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Organo ufficiale della Divisione SSOG di Innovhub Stazioni Sperimentali per l’IndustriaAzienda Speciale della Camera di Commercio di Milano

APRILE/GIUGNO 2013 ISSN 0035-6808 RISGARD 90 (2) 65-136 (2013)Poste Italiane S.p.a. - Spedizione in Abbonamento Postale -70% Finito di stampare nel mese di Giugno 2013 RISG

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GRAFICA IMPAGINAZIONE E STAMPA

LASERGRAFICA POLVER SRLVia Kramer, 17 - 20129 Milano

Redazione: [email protected] web: www.ssog.it

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biblioteca ssog it

Application note. Assessment of the cleaning performances ofhand dishwashing detergents. Bowl wash procedureDegradazione dei composti fenolici durante la conservazionedell’olio extra vergine di olivaSingle step extraction and derivatization of meat lipids for fattyacid Ultra Fast GC analysis

Seeds and seed oil compositions of Aleppo Pine (Pinus halepensis Mill.) grown in JordanInteresterification of goose fat and rapeseed oil mixture usingCandida rugosa lipase immobilized in alginate beads

Evaluation of fatty acids and physicochemical characteristics ofsix varieties of bambara groundnut (Vigna subterranea L. Verdc)seed oilsNota tecnica. Lubrificanti. Corrispondenze tra metodi analitici(gennaio-dicembre 2012)

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A P R I L E / G I U G N O 2 0 1 3 – A N N O X C

Sommario

D. Mariani, G. Pallotti, E. TrimignoM. Migliorini, C. Cherubini, L. Cecchi, B. ZanoniA. Ficarra, E.L. De Paola, D.P. Lo Fiego, G. Minelli, A. AntonelliS.K. Tukan, K. Al-Ismail, R.Y. Ajo,M.M. Al-DabbasK. Tarnowska, M. Kostecka, A. Piotrkowicz, M. Łobacz, B. KowalskiM.O. Aremu, S. Mamman, A. Olonisakin

M. Sala, F. Taormina, P. Fornasari, P. Ruggeri Notiziario

ORGANO UFFICIALE DELLA DIVISIONE SSOG DI INNOVHUB STAZIONI SPERIMENTALI PER L’INDUSTRIA

AZIENDA SPECIALE DELLA CAMERA DI COMMERCIO DI MILANO

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A. FABERI: Ministero delle Politiche Agricole Alimentari e Forestali – Roma

La RIVISTA ITALIANA DELLE SOSTANZE GRASSE è l’organo ufficiale della Divisione SSOG di Innovhub

Stazioni Sperimentali per l’Industria Azienda Speciale della

Camera di Commercio di Milano. Ha periodicità trimestrale e la scientificità dei contenuti è garantita

da un Comitato Internazionale di Referee.Pubblica lavori originali e sperimentali di autori

italiani ed esteri riguardanti la chimica, la biochimica, l’analisi e la tecnologia nei settori:

sostanze grasse e loro derivati, tensioattivi, detersivi, cosmetici, oli minerali.

Include metodi di analisi in inchiesta pubblica relativi ai settori di competenza, il Notiziario con

informazioni su congressi, notizie in breve e libri.La Rivista viene consultata in Italia dalle industrieproduttrici di oli e grassi alimentari ed industriali,

dalle industrie chimiche, da laboratori di enti statali,da istituti di ricerca e facoltà universitarie. E’ inoltre

distribuita all’estero in vari Paesi come Spagna,Grecia, Francia, Germania, Tunisia, Nigeria, Congo, Polonia, Romania, Bulgaria, Russia,

Stati Uniti, Brasile, Cina, Giappone, presso industrie produttrici ed esportatrici, enti pubblici e università,

da dove provengono diversi lavori scientifici.


L A R I V I S T A I T A L I A N A D E L L E S O S T A N Z E G R A S S E - V O L . X C - A P R I L E / G I U G N O 2 0 1 3

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A testing procedure for the assessment of the performances of hand dishwashing productsis described in this application note. Basing on the experiences already present into thescientific international landscape a simple but really reliable procedure was developed,also for making comparisons among different products or formulas.

Valutazione delle prestazioni di detergenti per il lavaggio manuale delle stoviglie. Procedura tipo Bowl-washIn questa nota applicativa viene descritta una procedura di verifica per la valutazione delleprestazioni dei prodotti per il lavaggio manuale dei piatti. Sulla base delle esperienze giàpresenti nel panorama scientifico internazionale, è stata sviluppata una procedura sempli-ce ma molto affidabile, adatta anche per la realizzazione di confronti tra diversi prodotti oformule.

D. Mariani*G. Pallotti

E. Trimigno

INNOVHUB - SSIAzienda Speciale della Camera di Commercio

Divisione SSOG - Milano

*CORRESPONDING AUTHOR:Dr. Mariani Davide

INNOVHUB - Divisione SSOGVia Giuseppe Colombo, 79

20133 Milano (Italy)E-mail: [email protected]

Application noteAssessment of the cleaning

performances of handdishwashing detergents.

Bowl wash procedure


1. INTRODUCTION

Despite of the general increasing, especially in theEuropean and North American society, of automat-ic procedures for cleaning the dishes above all dueto the greater penetration of dishwashers in themarket of the cited areas, (in Germany in 2010,67% of the households own a dishwashingmachine, 47% more than in 1980 [1], in Canada,the penetration rate for automatic dishwashers hasrisen from 54% in 1993 up to 70% in 2007 [2]) handdishwashing detergents are still considered of agreat importance on the market and in the develop-ment plains of many industries.This kind of products are water solution character-ized by a quite simple formulation in which, togeth-er with the mixture of surfactants responsible of thereal cleaning power of the product, is present acertain quantity of some other ingredients whichhave the task to make the product more apprecia-ble, like colorants, perfumes and additives for theadjustment of the viscosity as also the hedonisticaspect is became very important in the last years.The great part of the development’s efforts in thisfield are been done in the direction of a more widereliability with the most appreciated principles ofsustainability, in order to being able to give to thecustomers products more “green” but with thesame performance of the ones they are accus-tomed to used.The effort of changing the product lettingunchanged the cleaning performances is the rea-son why also the assessment of the performance ofthe hand dishwashing product is to be consideredan important activity, connected with a generaldevelopment towards sustainability.The methodology here below described is beendeveloped by the Detergents and SurfactantsDepartment of the SSOG division, a part ofInnovhub-SSI, Special Agency of the Chamber ofCommerce of Milan, starting from the examplesand the experiences already present into the scien-tific landscape related to the compartment of handdish washing products. [3]

2. DESCRIPTION OF THE METHOD

2.1 TEST PRINCIPLESSoiled plates are washed by hand in a dishwash-ing solution prepared using the products to be test-ed with a certain dilution, agreed with the client.

2.2 PREPARATION FOR THE EXECUTION OF THE TESTFor carrying out the test is necessary to prepareboth soiled plates and washing solution.

For soling the plates is necessary to prepare thesoiling mixture.

2.3 SOILING MIXTURE PREPARATION

Instructions:All components are separately weighted.Premix oleic acid and seeds oil melting them. Add wheat starch (or plain flour) to 1/3 of the water(demineralised) pre-heated at 60°C stirring byusing a mixer till obtaining a smooth paste.Add the premix to the hot water with flour stirringby using a mixer to a smooth paste. Add to themix (stirring) the rest of the water (demineralised)pre-heated at 100°C. Half of the soil is left boiling(low heat) for about 2 minutes, stirring. Add the other half part of the soil and let it cool atroom temperature, without forcing the process(minimum of three hours are necessary). When soil is stored overnight, seal the containerand keep it cool in the fridge.This soil can be stored in freezer up to one month.

2.4 SOILING PHASE:In order to have clean plates, wash them using acommon automatic dishwasher (normal program -Temperature 50-65°C)Put the first dry and cold plate on balance.Use disposable pipettes to dispense the amountof soil (5 g ± 0.1 g ) on each plate.Spread the soil evenly over the front surface ofeach plate and age the soil at least 10 minutes(letting it drying on the surface).Stack the plates at the side of the washing-upbasin.

2.5 EXECUTION OF THE TESTPut the required amount (g) of product directly inthe washing-up basin.Put the washing-up basin on the floor and add thewater at the required temperature using a reservoirpositioned with the outlet tap at a fixed height, (atleast 70 cm above the floor). This generates the initial lather.It is possible to agree with the customer the quan-tity of water to be used - from 2.5 to 5 liters - andthe hardness of the same water which has to beused for carrying out the test. Water is, in any case, tap water, hardened or soft-


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ened as necessary using appropriate devices.Immerse the first plate and wash it, using a plasticwashing up brush and making circular movementson all surface. Execute 20 circular complete move-ments on front surface, then turn the plate andmake other 6 movements on the back. Remove the washed plate from the bowl, let itdrain completely and pass to the next plate.Continue with the test, washing plates in thedescribed way, till reaching the end-point.

2.6 THE END POINTThe end-point is reached when the first permanentbreak in the foam layer covering the surface of thewashing solution is noted.The results are expressed in terms of number ofwashed dishes or of g of soil removed (5 g foreach dish washed).Carry out at least 4 replications of the test for eachsamples.These results can be statistically analyzed todetermine differences using, for example, statisti-cal methods for demonstrate, with a one-sided95% confidence range, that one of the testedproduct is as good as or better than another of thetested product.

3. EXPERIMENTAL

Comparison of 4 products following the lines of themethod (Tab. II, Tab. III).

Samples tested: 4 commercial products, here indi-cated as:Sample ASample BSample CSample D

Starting temperature of the solution: 45°CDosage: 4.5 gAmount of the washing solution: 3 litersConcentration: 1.5 g/l

3.1 STATISTICAL ELABORATION [4] (Tab. IV, Tab. V)Assessment of the significance of the differencesamong product applying a one-sided 95% confi-dence range.

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The p_values related to the comparison A>C, A>D,B>C and B>D are significative, with values lowerthen 0.05 (95% of confidence required).

In conclusion, in this case, products A and B areequal and better than product C and D which areequivalent (Tab. VI).

4. FINAL DISCUSSION

In order to check the stability of the procedure dur-ing the execution of the test it is possible to carryout some proofs using a reference product. In par-ticular, if the study is intended to last for severaldays, it could be considered a careful behavior toestablished the conformity of the procedure, dayby day, using the reference product which resultsare to be similar to the ones obtained during previ-ous days.For more, a randomization of the proofs with thevarious products which have to be tested is recom-mended.It has to be clear that the end point chosen, the firstpermanent break into the foam layer covering thesurface of the washing solution, is not the final

point in which the product loose completely its effi-cacy, but only a clear, identifiable point in which tostop the cleaning operation and the test in general.It is possible that the two events correspond butthis is not true unequivocally.The methodology described, in conclusion, let theoperator to proceed with the assessment of thewashing performances of the product that are to betested making a comparison among the variousend point determined using a statistical devicewhich permit the elaboration of the relativeassessed levels of efficacy.

BIBLIOGRAPHY

[1] ZVEI, 2011[2] OEE, 2007[3] Cg. Nitsch, G. Huttmann “Recommendation

for the quality assessment of the Cleaningperformances of hand dishwashing deter-gents”, IKW working group, SOFW-Journal,128 Jahrgang (5-2002)

[4] F. Pesarin, “Multivariate permutation test withapplication in Biostatistics” - John Wiley andSons 2001- Chichester

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L’olio extra vergine di oliva è un prodotto di particolare complessità. Il controllo della sua filieraproduttiva necessita conoscenze dei fenomeni di trasformazione della componente chimica e, diconseguenza anche funzionale, dell’olio. Durante lo stoccaggio e la distribuzione degli oli preval-gono le degradazioni ossidative non enzimatiche dei trigliceridi e le degradazioni idrolitiche deisecoiridoidi, che portano alla liberazione di idrossitirosolo e tirosolo. Scopo del lavoro è stato quello di seguire l’evoluzione in conservazione dei secoiridoidi e deriva-ti su oli differenti sia per il loro contenuto fenolico che per le differenti condizioni sperimentali distoccaggio dell’olio. Durante le campagne olearie 2008, 2009 e 2010 olive della cultivar Frantoiosono state lavorate in un frantoio operante a basso impatto ossidativo, in quanto basato sull’usodi una gramolatrice verticale che opera a 20°C in assenza d’aria. Gli oli prodotti durante la cam-pagna olearia 2008 e 2009 sono stati conservati in modo da simulare le normali condizioni diesposizione del prodotto in un punto vendita. Gli oli prodotti durante la campagna olearia 2010sono stati conservati in modo da accelerare le reazioni degradative. Al tempo zero e durante laconservazione, sui campioni di olio sono state misurati i parametri qualitativi di legge e il conte-nuto dei composti fenolici per HPLC secondo il metodo COI.I campioni di olio erano tutti a bassa acidità e con contenuti simili di acido oleico; un campionesu tre è risultato invece diverso per l'intensità di amaro e per il contenuto totale di composti feno-lici. Tutti i campioni erano poco degradati ovvero a basso contenuto dei parametri espressionedella degradazione ossidativa dei trigliceridi e della degradazione idrolitica dei secoiridoidi. Durante le prove di conservazione tutti i campioni di olio hanno subito una significativa degrada-zione sia in termini assoluti che in relazione al tempo zero di conservazione. I risultati hannodimostrato la significatività dei composti forma dialdeidica del decarbossimetil oleoeuropeinaaglicone (3,4 DHPEA-EDA) e idrossitirosolo (3,4 DHPEA) come indicatori della degradazioneidrolitica dei composti fenolici: il primo in relazione inversa alla degradazione, il secondo in rela-zione diretta con la degradazione. La ricerca ha infine evidenziato l’effetto protettivo dei compostifenolici sull’autossidazione dei trigliceridi.

Degradation of phenolic compounds during extra virgin olive oil shelf-lifeExtra virgin olive oil is a particularly complex product. Controlling the relevant production chainrequires having a good knowledge of both transformation phenomena occurring in its chemicalcomponent and, as a result, functional changes in oil. During oil storage and distribution bothnon-enzymatic oxidative degradation of triglycerides and hydrolytic degradation of secoiridoidsprevail, leading to the formation of hydroxytyrosol and tyrosol. The aim of this work was to follow the evolution during storage of secoiridoids and derivatives indifferent oil types, having different phenol contents and stored under different experimental con-ditions. During 2008, 2009, and 2010 oil crop seasons, olives from Frantoio cultivar wereprocessed in an oil mill working in low oxidative impact conditions. Oil produced during the 2008and 2009 crop seasons was stored to simulate normal shelf-life conditions of products in a store.Oil produced during the 2010 crop season was stored under specific conditions to acceleratedegradation reactions. Legal quality parameters and phenol compound content were measuredon olive oil samples by HPLC according to the COI method at both time zero and during storage.All the oil samples had a low acidity value and a similar oleic acid content. One out of three oilsamples resulted to be different in bitter intensity and total phenol compound content. All the oilsamples were subjected to low degradation, namely they showed a low parameter content, whichwas indicative of either oxidative degradation of triglycerides or hydrolytic degradation of secoiri-doids. During storage trials, all the oil samples were subjected to significant degradation both inabsolute terms and in relation to storage time zero.Results showed the significance of compounds dialdehydic form of decarboxymethyl oleuropeinaglycone (3,4 DHPEA-EDA) and hydroxytyrosol (3,4 DHPEA) as indicators of hydrolytic degra-dation of phenol compounds, the former being inversely related to degradation, while the latterwas directly related to degradation. Our research demonstrated a protective effect of phenol com-pounds on autoxidation of triglycerides.

Degradazione dei composti fenolici durante la conservazione dell’olio

extra vergine di oliva

M. Migliorini1

C. Cherubini1

L. Cecchi1

B. Zanoni2*

1Laboratorio ChimicoMerceologico,

Agenzia Speciale della Cameradi Commercio - Firenze

2Dipartimento di BiotecnologieAgrarie, Sezione

di Tecnologie Alimentari,Università degli Studi di Firenze

*AUTORE DI RIFERIMENTOProf. Bruno Zanoni

e-mail: [email protected].: +39 055 3288474Fax: +39 055 3288480


INTRODUZIONE

L’olio extra vergine di oliva è un prodotto di partico-lare complessità. Il controllo della sua filiera pro-duttiva necessita conoscenze non solo dei feno-meni puramente estrattivi dell’olio contenuto nelleolive, ma anche dei fenomeni di trasformazionedella componente chimica e, di conseguenzaanche funzionale, dell’olio [1, 2]. Mentre le trasformazioni a carico degli acidi grassisono note (i.e. idrolisi enzimatica dei trigliceridi [3],autossidazione e fotossidazione dei trigliceridi edegli acidi grassi liberi [4, 5], ossidazione enzima-tica degli acidi grassi liberi linoleico e linolenico o“LOX pathway” [6]), meno comprese sono le tra-sformazioni a carico della classe dei compostifenolici caratteristici delle olive, i secoiridoidi.Si ritiene che i composti primari presenti nelle olivemature, oleoeuropeina e ligstroside, possano subiresia una trasformazione idrolitica che una trasforma-zione di natura ossidativa [1, 7-11]. Tali trasformazio-ni hanno come effetto la variazione negli oli sia del-l'intensità di descrittori sensoriali quali l'amaro e ilpiccante [12, 13] che l'entità del potere antiossidan-te, con conseguente variazione della shelf-life e delvalore nutrizionale del prodotto [14, 15].L’ipotesi è che, per effetto della degradazione idro-litica, si formino gli agliconi (3,4-DHPEA-EA - oleo-europeina aglicone; p-HPEA-EA - ligstroside agli-cone) a seguito della perdita, dovuta all'azionedella β-glucosidasi, della molecola di zucchero; gliagliconi si idrolizzino poi a dare le relative formedialdeidiche e queste ultime si idrolizzino nei rela-tivi carbossimetil agliconi (3,4-DHPEA-EDA - formadialdeidica del decarbossimetil oleoeuropeinaaglicone; p-HPEA-EDA - forma dialdeidica deldecarbossimetil ligstroside aglicone). Infine secon-do meccanismi idrolitici ancora poco conosciuti siarriva a quelli che sono i composti finali di idrolisi:l'idrossitirosolo (3,4- DHPEA) e il tirosolo (p-HPEA). Per quanto riguarda la degradazione ossidativadei secoiridoidi si ipotizza prima di tutto una degra-dazione di natura enzimatica: le polifenolossidasi(PPO) e le perossidasi (POD) presenti nelle pasted'oliva catalizzano in presenza di ossigeno l'ossi-dazione dei composti fenolici a dare chinoni (i.e. ilcambiamento di colore verso il bruno delle paste inlavorazione è prova di tale ossidazione). Si ipotizzaanche un'ossidazione di natura non enzimatica,legata alle reazioni di terminazione dell'autossida-zione radicalica dei trigliceridi a dare perossidi ederivati: la cessione di atomi di idrogeno da partedei composti fenolici blocca la formazione dei radi-cali idroperossido. Si è notato come le suddette degradazioni deisecoiridoidi avvengano in momenti diversi del pro-cesso di produzione dell'olio extravergine. Durantele fasi di estrazione, in particolare durante la fran-

gitura e la gramolatura, prevalgono le degradazio-ni ossidative di natura enzimatica e le degradazio-ni idrolitiche che portano alla formazione delleforme dialdeidiche dei carbossimeti agliconi [16,17]. Durante lo stoccaggio e la distribuzione deglioli prevalgono invece le degradazioni ossidativenon enzimatiche e le degradazioni idrolitiche cheportano alla formazione di idrossitirosolo e tirosolo[11, 18, 19]. A tale proposito Siliani et al. [20]hanno costruito un modello concettuale di previsio-ne della conservabilità dell’olio extra vergine dioliva basato sul contenuto dei composti fenolici: laconcentrazione dell’oleoeuropeina aglicone comeindice favorevole per la stabilità e correlato diretta-mente con l’intensità di amaro dell’olio, la concen-trazione di idrossitirosolo come parametro espres-sione della degradazione dell’olio durante la shelf-life.Scopo di questo lavoro è stato quello di seguirel’evoluzione in conservazione dei secoiridoidi ederivati su oli differenti sia per il loro contenutofenolico che conservati in differenti condizioni spe-rimentali.

MATERIALI E METODI

DISEGNO SPERIMENTALEDurante le campagne olearie 2008, 2009 e 2010olive sane e al giusto grado di maturazione dellacultivar Frantoio sono state lavorare entro 4 oredalla raccolta in un frantoio (Toscana EnologicaMori, Firenze), messo in opera presso l’AziendaAgricola di Torre Bianca a San Casciano Val diPesa (Firenze). Il frantoio era dotato di una gramo-latrice verticale operante a basso impatto ossidati-vo, in quanto in grado di operare a 20°C in assen-za d’aria come già descritto da Migliorini et al. [21].Al termine dell’estrazione in decanter a due fasi glioli sono stati filtrati mediante un filtro pressa. I lottiomogenei di oli extra vergine di oliva così ottenutisono stati sottoposti a prove di conservabilità.Gli oli prodotti durante la campagna olearia 2008(i.e. oli extra vergine tipo A) e 2009 (i.e. oli extravergine tipo B) sono stati conservati in bottiglieambrate da 40 ml e tappate con apposito tappo avite (Unilab, Gravina di Catania, Catania); le botti-glie sono state conservate sotto luce artificiale fluo-rescente (8 ore al giorno), in condizioni di tempera-tura controllata a 18±2°C, così da simulare le nor-mali condizioni di esposizione del prodotto in unpunto vendita. L’intensità della luce, misurata conun luminometro portatile (modello HD 2302.0, DeltaOhm rsl, Padova), era di circa 380 lux. Al tempozero e approssimativamente ogni tre mesi di con-servazione, gli oli sono stati analizzati in triplo. Ladurata della sperimentazione è stata di circa 14mesi.Gli oli prodotti durante la campagna olearia 2010

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(i.e. oli extra vergine tipo C) sono stati conservati inbottiglie di vetro chiaro da 40 ml, tappate conapposito tappo a vite (Unilab, Gravina di Catania,Catania); le bottiglie sono state conservate allatemperatura di 60°C in una cella termostatata(mod. Nse60, SARIN s.a.s., Firenze), per accelera-re le reazioni degradative. Al tempo zero e appros-simativamente ogni 15 giorni di conservazione, glioli sono stati analizzati in triplo. La durata dellasperimentazione è stata di circa 4 mesi.

DETERMINAZIONE ANALITICHE• Le determinazioni del contenuto in acidità, del

numero di perossidi e delle misure spettrofoto-metriche sono state eseguite in accordo allemetodiche ufficiali riportate nel Reg. CEE2568/1991 [22]. La determinazione della com-posizione acidica è stata eseguita in accordoalla metodica ufficiale riportata nel Reg. CE796/2002 [23].

• La determinazione del contenuto in compostifenolici e la loro identificazione è stata eseguitaper HPLC in accordo con il metodo ufficiale COI[24]. L’estrazione è stata condotta usando unasoluzione metanolo/acqua (80:20); i compostifenolici estratti sono stati separati e, quindi,determinati con il cromatografo liquido HewlettPackard mod. 1200 Diode-Array Detector, muni-to di autocampionatore Hewlett Packard mod.1200 (Agilent Technologies, Santa Clara, Califor-nia, USA). Le condizioni di analisi sono state leseguenti. Colonna HPLC: LiChrospher 100endcapped RP-18, 5 μm, 250 × 4,6 mm DI; volu-me di iniezione: 20 μl; solvente: acqua con lo0,2% di H3PO4/acetonitrile/metanolo in gradien-te [24]; lunghezza d’onda: 280 nm.

• L’analisi quantitativa è stata effettuata utilizzando

l’acido siringico come standard interno e il tiro-solo come composto di riferimento per il fattoredi risposta. I derivati naturali e ossidati di oleuro-peina e ligstroside (i.e. secoiridoidi), i lignani, iflavonoidi e gli acidi fenolici sono stati quantifi-cati in mgtirosolokgolio-1. Il contenuto totale di com-posti fenolici (mgtirosolokgolio-1) è stato determina-to dalla somma delle aree dei picchi registrati a280 nm.

• L’analisi sensoriale è stata eseguita in accordocon la metodica comunitaria riportata nel Rego-lamento CE 640/2008 [25].

RISULTATI E DISCUSSIONE

Tutti gli oli al termine del loro processo di produzio-ne erano, sia dal punto di vista chimico-fisico chesensoriale, extra vergini di oliva in accordo alRegolamento CE 796/2002 [23]. La Tabella I riporta la relativa composizione deicampioni di olio, in cui sono state esplicitate lecaratteristiche che secondo il modello di Siliani etal. [20] sono indici di stabilità (i.e. in modo diretto oinverso favoriscono la stabilità in conservazione) oparametri di degradazione (i.e. esprimono il gradodi degradazione in conservazione). I campioni diolio erano tutti a bassa acidità e con contenuti simi-li di acido oleico; un campione su tre è risultatoinvece diverso per l'intensità di amaro e per il con-tenuto totale di composti fenolici. Tutti i campionierano poco degradati ovvero a basso contenutodei parametri espressione della degradazioneossidativa dei trigliceridi e della degradazione idro-litica dei secoiridoidi, a conferma delle buone con-dizioni operative di processo adottate per minimiz-zare la degradazione. La tabella II riporta la concentrazione dei singoli

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composti fenolici. A conferma di quanto riportato inletteratura [26], il componente quantitativamentepiù significativo è stato la forma dialdeidica deldecarbossimetil oleoeuropeina aglicone (3,4-DHPEA-EDA), la cui formazione è espressionedelle rapide degradazioni idrolitiche a carico deisecoiridoidi nel corso del processo di estrazionedell’olio. Coerentemente al contenuto totale dicomposti fenolici, gli oli di tipo B sono stati i cam-pioni a minore contenuto di 3,4-DHPEA-EDA.E’ anche interessante sottolineare come il contenu-to di aglicone oleoeuropeina sia risultato proporzio-nale all’intensità del descrittore amaro dei campio-ni di olio analizzati. Applicando infatti il modelloempirico esponenziale di Siliani et al. [12]

y = 7,46 · exp (0,716 · x)

e convertendo proporzionalmente le misurateintensità di amaro (x) dalla scala 0-10 alla scala 0-5 (come riportata nel modello), il valore 3,3 (1,7nella scala modificata) di amaro degli oli di tipo Aha portato a prevedere un contenuto di agliconeoleuropeina (y) di 25 mg/kg, mentre il valore 4 (2nella scala modificata) di amaro degli oli di tipo Be C ha portato a prevedere un contenuto di aglico-ne oleuropeina di 31 mg/kg; valori previsti dellostesso ordine di grandezza di quelli misurati: 27,

47 e 49 mg/kg rispettivamente per gli oli A, B e C.Le Tabelle III-VI riportano le variazioni dei parame-tri di degradazione degli oli durante le prove diconservazione.E’ evidente per tutti i campioni di olio una significa-tiva degradazione ossidativa dei trigliceridi, tale daportare gli oli a non poter essere legalmente consi-derati extra vergini (Tab. III). Si è avuto in modocoerente un incremento del numero di perossidi,del K232 in quanto espressione dei dieni coniugatipresenti negli idroperossidi e del K270 in quantoespressione dei prodotti secondari dell’irrancidi-mento ossidativo [18]. Nel caso degli oli conserva-ti alla luce ed a temperatura ambiente la non con-formità legale è avvenuta solo dopo più di 12 mesidi conservazione, mentre per gli oli sottoposti adegradazione accelerata a 60°C la non conformitàlegale si è raggiunta dopo circa 3 mesi. In unasimile sperimentazione condotta in condizioni didegradazione accelerata a 60°C da Lerma-Garciaet al. [11] gli oli hanno invece superato i limiti qua-litativi legali già dopo una settimana di conserva-zione; ciò è stato probabilmente dovuto ad un con-tenuto fenolico totale che era dalle 2 alle 3 volte piùbasso di quello misurato negli oli da noi prodotti.Analizzando i composti fenolici si è osservata unacomplessiva diminuzione della loro concentrazione(Tab. IV-VI). Durante la conservazione, per tutti

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i campioni di olio, i componenti che hanno presen-tato un andamento in grado di esprimere meglio ilfenomeno degradativo sono stati le forme dialdei-diche dei decarbossimetil oleoeuropeina e ligstro-side agliconi (3,4 DHPEA-EDA, p-HPEA-EDA),l’idrossitirosolo (3,4 DHPEA) e tirosolo (p-HPEA). Inrelazione ai dati di letteratura [11, 19, 27] è ipotiz-zabile una degradazione idrolitica nel tempo deicomposti fenolici quantitativamente più rappresen-tativi con conseguente incremento dei compostifinali di idrolisi, idrossitirosolo e tirosolo.Si sono successivamente confrontate le velocità didegradazione per le tre tipologie di olio. Sono statecalcolate le variazioni relative rispetto al tempozero di conservazione dei parametri di degradazio-ne, risultati dai dati precedenti quantitativamente

più interessanti: il numero di perossidi, il K232, leconcentrazioni di 3,4 DHPEA-EDA e 3,4 DHPEA.Le Figure 1-3 riportano gli andamenti dei relatividati sperimentali, descritti, per puri motivi di chia-rezza espositiva, con significative linee di tenden-za lineari o esponenziali. Si è evidenziata una netta differenza di velocità didegradazione dei campioni di olio. I campioni ditipo C, essendo stati sottoposti ad un test accele-rato di conservazione a 60°C, sono quelli a mag-giore velocità di degradazione ossidativa dei trigli-ceridi e idrolitica dei composti fenolici. La differen-za di velocità tra i campioni di tipo A e B è secon-do noi invece espressione del differente contenutoiniziale in composti fenolici (Tab. I). Coerentemen-te con il modello di Siliani et al. [20], i campioni di

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tipo B, a minore contenuto di composti fenolici,hanno avuto una degradazione sia ossidativa cheidrolitica più rapida dei campioni di tipo A, a mag-giore contenuto di composti fenolici. Un analogocomportamento è stato evidenziato da Lerma-Gar-cia et al. [11] dal confronto di oli con un contenutofenolico di circa 200 mg/kg rispetto agli stessi oliprivati della componente fenolica mediante estra-zione liquida-liquida in laboratorio.E’ interessante osservare come per tutti i campionidi olio ad una diminuzione tendenzialmente linearedella concentrazione della forma dialdeidica deldecarbossimetil oleoeuropeina aglicone è corri-sposto coerentemente un incremento tendenzial-mente lineare dell’idrossitirosolo (Fig. 3).

CONCLUSIONI

Il disegno sperimentale adottato in questa ricercaha permesso di ottenere lotti di olio omogenei alloro interno, estratti in campagne olearie successi-ve con identiche condizioni operative di processo,in grado di minimizzare non conformità legali chi-mico-fisiche e sensoriali del prodotto.La qualità iniziale delle tre tipologie di olio e le loromodalità di conservazione hanno permesso il con-fronto tra due campioni di olio (tipo A e B) a diver-sa suscettibilità per la degradazione, in quanto adiverso contenuto di composti fenolici antiossidan-ti, e un campione di olio (tipo C) conservato in untest accelerato a 60°C da intendersi come riferi-mento.I risultati hanno dimostrato la significatività deicomposti forma dialdeidica del decarbossimetiloleoeuropeina aglicone (3,4 DHPEA-EDA) e idros-sitirosolo (3,4 DHPEA) come indicatori della degra-dazione idrolitica dei composti fenolici: il primo inrelazione inversa alla degradazione, il secondo inrelazione diretta con la degradazione. La ricerca ha infine evidenziato il reale effetto pro-tettivo dei composti fenolici sull’autossidazione deitrigliceridi, a supporto del modello concettuale diprevisione della conservabilità dell’olio extra vergi-ne di oliva messo a punto da Siliani et al. [20].

RINGRAZIAMENTI

Un sentito ringraziamento al Prof. AlessandroParenti del Dipartimento di Economia, Ingegneria,Scienze e Tecnologie Agrarie e Forestali per lamisura delle lunghezze d’onda della luce fluore-scente.

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qualità, Camera di Commercio, Firenze (2005). [2] L. Di Giovacchino, S. Sestili, D. Di Vincenzo,

Influence of olive processing on virgin olive oilquality, European Journal of Lipid Scienceand Technology 104, 587-601 (2002).

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[4] M.T. Morales, J.J. Rios, R. Aparicio, Changesin the volatile composition of virgin olive oilduring oxidation: flavors and off-flavors, Jour-nal of Agricultural and Food Chemistry 45,2666-2673 (1997).

[5] E.N. Frankel, Recent advances in lipid oxida-tion, Journal of the Science of Food and Agri-culture 54, 495-511 (1991).

[6] F. Angerosa, R. Mostallino, C. Basti, R. Vito,Virgin olive oil odour notes: their relationshipswith volatile compounds from the lipoxyge-nase pathway and secoiridoid compounds,Food Chemistry 68, 283-287 (2000).

[7] F. Gutierrez-Rosales, M.P. Romero, M.Casanovas, M.J. Motilva, M.I. Mìnguez-Mos-quera, Metabolites involved in Oleoeuropeinaccumulation and degradation in fruits ofOlea europea L.: Hojiblanca and Arbequinavarieties, Journal of Agricultural and FoodChemistry 58, 12924-12933 (2010).

[8] L.S. Artajo, M.P. Romero, M. Suàrez, M.J.Motilva, Partition of phenolic compounds dur-ing the virgin olive oil industrial extractionprocess, European Food Research and Tech-nology 225, 617-625 (2007).

[9] M. Servili, R. Selvaggini, S. Esposto, A. Tatic-chi, G.F. Montedoro, G.Morozzi, Health andsensory properties of virgin olive oilhydrophilic phenols: agronomic and techno-logical aspects of production that affect theiroccurence in the oil, Journal of Chromatogra-phy A. 1054, 113-127 (2004).

[10] P. Rovellini, N. Cortesi, Liquid chromatogra-phy-mass spectrometry in the study of oleu-ropein and ligstroside aglycons in virgin oliveoil: aldeydic, dialdeydic forms and their oxi-dized products, Riv. Ital. Sostanze Grasse 79,1-13 (2002).

[11] M.J. Lerma-Garcia, E.F. Simo-Alfonso, E. Chi-avaro, A. Bendini, G. Lercker, L. Cerretani,Study of chemical changes produced in virginolive oils with different phenolic contents dur-ing an accelerated storage treatment, Journalof Agriculture and Food Chemistry 57, 7834-7840 (2009).

[12] S. Siliani, A. Mattei, B. Zanoni, L. BenevieriInnocenti, Bitter taste and phenolic com-pounds in extra virgin olive oil: an empiricalrelationship, Journal of Food Quality 29, 431-441 (2006).

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[13] P. Andrewes, J.L.H.C. Busch, T. De Joode, A.Groenewegen, H. Alexandre, Sensory proper-ties of virgin olive oil poliphenols: identifica-tion of deacetoxy-ligstroside aglycon as a keycontributor to pungency, Journal of Agricultur-al and Food Chemistry 51, 1415-1420 (2003).

[14] G.K. Beauchamp, R.S.J. Keast, D. Morel, J.Lin, J. Pika, Q. Han, C. Lee, A.B. Smith, P.A.S.Breslin, Ibuprofen-like activity in extra-virginolive oil, Nature 437, 45-46 (2005).

[15] F. Visioli, G. Bellomo, C. Galli, Free radicalscavenging properties of olive oil polyphe-nols, Biochemical and Biophysical ResearchCommunications 247, 60-64 (1998).

[16] M. Servili, A. Taticchi, S. Esposto, S. Urbani,R. Selvaggini, G. Montedoro, Influence of thedecrease in oxygen during malaxation of olivepaste on the composition of volatiles and phe-nolic compounds in virgin olive oil, Journal ofAgricultural and Food Chemistry 56, 10048-10055 (2008).

[17] C.M. Kalua, D.R. Bedgood, A.G. Bishop, P.D.Prenzler, Changes in volatile and phenoliccompounds with malaxation time and temper-ature during virgin olive oil production, Jour-nal of Agricultural and Food Chemistry 54,7641-7651 (2006).

[18] A. Bendini, L. Cerretani, S. Vecchi, A. Carras-co-Pancorbo, G. Lercker, Protective effects ofextra virgin olive oil phenolics on oxidativestability in the presence or absence of copperions, Journal of Agricultural and Food Chem-istry 54, 4880-4887 (2006).

[19] M. Brenes, A. Garcia, P. Garcia, A. Garrido,Acid Hydrolysis of secoiridoid aglycons dur-ing storage of virgin olive oil, Journal of Agri-cultural and Food Chemistry 49, 5609-5614(2001).

[20] S. Siliani, B. Zanoni, G. Fia, M. Bertuccioli, A.Mattei, O. Lorenzini, Verso un modello concet-tuale di previsione della conservabilità del-l’olio extra vergine di oliva, Riv. Ital. SostanzeGrasse 96, 31-38 (2009).

[21] M. Migliorini, C. Cherubini, B. Zanoni, M.Mugelli, E. Cini, A. Berti, Influenza delle con-dizioni operative di gramolatura sulla qualitàdell’olio extra vergine di oliva, Riv. Ital. Sostan-ze Grasse 86, 92-102 (2009).

[22] Commissione Europea, Regolamento (CEE)n. 2568/91 della Commissione dell’11 lugliorelativo alle caratteristiche degli oli d’oliva edegli oli di sansa d’oliva nonché ai metodi adessi attinenti, GUCE, L248, 1-83 (1991).

[23] Commissione Europea, Regolamento (CE) n.796/2002 della Commissione del 6 maggio2002 recante modifica del regolamento (CEE)n. 2568/91 relativo alle caratteristiche degli olid’oliva e degli oli di sansa d’oliva nonché ai

metodi ad essi attinenti e le note complemen-tari di cui all’allegato del regolamento (CEE)n. 2658/87 del Consiglio relativo alla nomen-clatura tariffaria e statistica e alla tariffa doga-nale comune, GUCE, L128, 8-28 (2002).

[24] COI/T.20/Doc. n. 29, Metodi ufficiali di analisi.Determinazione dei biofenoli degli oli di olivamediante HPLC, Consiglio Oleicolo Interna-zionale, Madrid (Spagna) Novembre 2009.

[25] Commissione Europea, Regolamento (CE) n.640/2008 della Commissione del 4 luglio 2008che modifica il regolamento (CEE) n. 2568/91relativo alle caratteristiche degli oli d’oliva edegli oli di sansa d’oliva nonché ai metodi adessi attinenti, GUCE, L178, 11-16 (2008).

[26] M. Migliorini, L. Cecchi, C. Cherubini, S. Tra-pani, E. Cini, B. Zanoni, Understandingdegradation of phenolic compounds duringolive oil processing by inhibitor addition, Euro-pean Journal of Lipid Science and Technolo-gy, in fase di stampa (2012).

[27] A. Romani, C. Lapucci, C. Cantini, F. Ieri, N.Mulinacci, F. Visioli, Evolution of minor polarcompounds and antioxidant capacity duringstorage of bottled extra virgin olive oil, Journalof Agricultural and Food Chemistry 55, 1315-1320 (2007).

Degradation of phenolic compounds duringextra virgin olive oil shelf-lifeExtra virgin olive oil is a particularly complex prod-uct. Controlling the relevant production chainrequires a good knowledge of both transformationphenomena occurring in its chemical componentand, as a result, functional changes in oil. Duringoil storage and distribution both non-enzymaticoxidative degradation of triglycerides and hydrolyt-ic degradation of secoiridoids prevail, leading tothe formation of hydroxytyrosol and tyrosol.The aim of this work was to follow the evolution dur-ing storage of secoiridoids and derivatives in differ-ent oil types, having different phenol contents andstored under different experimental conditions. During 2008, 2009, and 2010 oil crop seasons,olives from Frantoio cultivar were processed in anoil mill working in low oxidative impact conditions.Oil produced during 2008 (i.e. extra virgin olive oiltype A) and 2009 crop seasons (i.e. extra virginolive oil type B) was stored in closed, ambercoloured bottles under fluorescent lamp light at18°C. Oil produced during 2010 crop season (i.e.extra virgin olive oil type C) was stored in clearglass bottles at a temperature of 60°C. Legal qual-ity parameters and phenol compound content weremeasured on olive oil samples by HPLC accordingto the COI method at both time zero and duringstorage.All the oil samples had a low acidity value and a

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similar oleic acid content. One out of three oil sam-ples resulted to be different in bitter intensity andtotal phenol compound content (Tab. I). All the oilsamples were subjected to low degradation,namely they showed a low parameter content,which was indicative of either oxidative degrada-tion of triglycerides or hydrolytic degradation ofsecoiridoids (Tab. II). The quantitatively most sig-nificant component of secoiridoids was the dialde-hydic form of decarboxymethyl oleuropein agly-cone (3,4-DHPEA-EDA), which forms as a result ofrapid hydrolytic degradation of secoiridoids duringoil extraction process. During storage trials, all the oil samples were sub-jected to significant oxidative degradation oftriglycerides in absolute terms (Tab. III). A consis-tent increase was observed in peroxide number,K232 as expression of conjugated dienes inhydroperoxides and K270 as expression of deriva-tives from oxidative rancidification. Phenol compounds exhibited an overall decreasein concentration (Tab. IV-VI). The best trend toexpress degradation phenomenon was shown byfollowing components: the dialdehydic form ofdecarboxymethyl oleuropein and ligstroside agly-cones (3,4 DHPEA-EDA, p-HPEA-EDA), hydroxyty-rosol (3,4 DHPEA) and tyrosol (p-HPEA). Hydrolyt-

ic degradation over time of the quantitatively mostrepresentative phenol compounds was assumed,with a resulting increase in final compounds ofhydrolysis, hydroxytyrosol and tyrosol.In addition, rates of degradation were comparedbetween the three different oil types. Relativechanges were calculated, as compared to storagetime zero, in degradation parameters: peroxidenumber (Fig. 1), K232 (Fig. 2), 3,4 DHPEA-EDA and3,4 DHPEA (Fig. 3). A marked difference in rate ofdegradation of oil samples was determined. Type C samples, which were subjected to anaccelerated storage test at 60°C, showed the high-est rate of oxidative degradation of triglyceridesand hydrolytic degradation of phenol compounds. The difference in rate between type A and B sam-ples resulted from a different initial phenol com-pound content. In agreement with literature data,type B samples, which had a low phenol com-pound content, showed more rapid oxidative andhydrolytic degradation than type A samples, whichhad a high phenol compound content. This demon-strated a protective effect of phenol compounds onautoxidation of triglycerides.

Received, February 10, 2012Accepted, March 26, 2012

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Two different methods for direct fat extraction of meat and fatty acid transesterification havebeen tested. Freeze-drying (FD) of the meat sample, with or without a previous enzymaticdigestion, followed by a base-catalyzed transmethylation (KOH/MeOH) was carried out.Both methods were compared with the standard Folch (FOL) procedure of fat extractionfollowed by the same transmethylation. The fatty acids composition was determined byUltra Fast GC. FD showed absolute quantities comparable with data obtained from fattyacid methyl ester (FAME) analysis starting from lipids obtained with the FOL procedure.In addition, FD allows a lower solvent disposal plus a considerable time saving.Keywords: Fatty acids analysis; Meat lipids; Single step transesterification; Fast GC;Rapid derivatization methods

Estrazione e derivatizzazione contemporanea dei lipidi della carne per l’ana-lisi degli acidi grassi con Ultra Fast GCSono stati confrontati 2 diversi metodi di estrazione del grasso dalla carne ed è stato testa-to il grado di trans esterificazione degli acidi grassi. Sono stati transmetilati tramite cata-lisi basica (KOH/MeOH) campioni liofilizzati di carne (FD), con o senza digestione enzi-matica preventiva. Entrambi i metodi sono stati confrontati con la procedura standard diFolch (FOL) per l’estrazione dei grassi ed è stata effettuata la stessa transmetilazione. Lacomposizione in acidi grassi è stata determinata usando un Ultra Fast GC. La FD hamostrato quantità assolute comparabili con i dati ottenuti dall’analisi dei metilesteri degliacidi grassi a partire dal grasso estratto con il metodo FOL. Inoltre la FD permette unminor spreco di tempo e di solventi.Parole chiave: Analisi degli acidi grassi; lipidi della carne; transesterificazione a singo-lo step; Fast GC; metodi di derivatizzazione rapida

Single step extraction andderivatization of meat lipids for fatty

acid Ultra Fast GC analysis

A. FicarraE.L. De Paola*

D.P. Lo FiegoG. Minelli

A. Antonelli

Department of Agricultural and Food Sciences

Faculty of Agriculture, University of Modena and

Reggio EmiliaPad. Besta, Reggio Emilia

*CORRESPONDING AUTHORFax: +39 522 522053

E-mail: [email protected]


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1. INTRODUCTION

The fatty acids (FA) composition gives a unique fin-gerprint that can be used as a quality indicatoragainst frauds in several food products. Moreover,an accurate estimation of fatty acids composition isvery important in defining the nutritional value andit could give an idea of the food history [1]. FAanalysis generally requires three steps:fat extraction; FA derivatization (usually as methyl esters - FAME); FAME GC analysis. This last step can be very fast. Short narrow-borecolumns are able to give perfect separations ofcomplex FAME mixtures in couples of minutes [2],and nowadays many fast GC systems are largelycommercialized. However, the full exploitation ofthese systems requires extraction and derivatiza-tion methods up to the task that fast GC equip-ments should deserve. In fact, to avoid spoiling theadvantage of a quick GC separation, a continuousstream of samples should be prepared by usingreliable methods. In a previous paper [3], the prob-lem of FAME preparation was solved through afive-min procedure transforming crude fat into aready-to-injection FAME solution. However, themost difficult and time demanding step was still thefirst one: fat extraction. This phase is always com-plex, time and solvent consuming. When aqueous-containing samples are involved, the proceduresbecome even more cumbersome. Folch, Lees,Sloane, & Stanley [4] in 1957 and Boselli et al. [10]in 2001 or Bligh & Dyer [5] in 1959 are two of themost popular examples.The aim of speeding up this step is a very demand-ing task. New techniques, such as supercriticalfluid extraction [6] or high-pressure solvent extrac-tion [7, 8] are of limited effectiveness. For meatsamples, the large amount of water is the core ofthe problem. Its elimination would make the extrac-tion more open to alternative solutions. Thus, itwould be possible to join fat extraction and FAderivatization in a single step, as it was successful-ly proposed by Conte et al. [9] in 1989 for sun-flower seeds. However, oily seeds are naturallyvery rich in fat (10-30%), while water content isalmost negligible if compared to meat. In addition,oil is regularly stored into the endosperm cells,making its extraction quite simple. Meat, on thecontrary, has only 1-5% of fat, which is closely con-nected to proteins and cell structures.Considering these characteristics of the meat sam-ples, the possibility of using a single-step proce-dure to extract and simultaneously derivatize FA isa very difficult and attractive goal. Previous resultsof this approach are reported in this short commu-nication. A preliminary evaluation of the single-stepprocedure was carried out by examining

KOH/MeOH derivatization directly applied tofreeze-dried meat samples. An approach to previ-ously enzymatically digested and then freeze-driedsamples was also verified. These early results werecompared with classical FOL extraction followedby fat derivatization, considered as the referenceprocedure.

2. MATERIALS AND METHODS

2.1 SAMPLE RETRIEVAL AND PREPARATIONThe meat sample used for the experimentation wastaken from muscle (Longissimus dorsi) of pork car-casses from the Mora Romagnola, an autochtho-nous Italian pig. The tissue (100 g) was ground ina piece of homemade equipment for 1 min.

2.2 MEAT DIGESTIONAn aliquot of ground meat (1 g) was transferredinto a 100 mm × 16 mm test tube with Teflon-linedcap (5 replicates for each method). An aliquot ofpapain solution (1% w/v; 2 ml) was added to thesample and it was incubated at 60°C for 4 h. Thenthe sample was frozen at -20°C.

2.3 FREEZE-DRYINGAn aliquot of ground meat (1 g) was transferred ina 100 mm × 16 mm test tube with Teflon-lined cap(5 replicates for each method). Tubes were frozenat -20°C and than they were placed in a rack insidea desiccator. Digested samples were treated in the same way.After connecting the desiccator to the freeze-drier,samples were set aside up to complete drying (24 h).

2.4 REAGENTSAll solvents and reagents were of analytical grade;chloroform, methanol, n-hexane, potassiumhydroxide, and enzyme (papain) were obtainedfrom Sigma-Aldrich (Milan, Italy).Single fatty acid standards were purchased fromLarodan Fine Chemicals AB (Malmö, Sweden),methyl nonadecanoate (C19:0) assay ≥ 99% as aninternal standard was purchased from Fluka (Milan,Italy) and prepared at a concentration of 1% (w/v)in hexane.

2.5 PREPARATION AND ANALYSIS OF FATTY ACIDMETHYL ESTERS2.5.1 Single stepThe freeze-dried samples were operated as Ficar-ra et al. (2010) have already described for fat, dou-bling the amount of each reagent. Strictly in thisorder, hexane (4 ml) and internal standard wereadded and mixed with an Ultra-Turrax (T25 basic,IKA, WERKE; max velocity for 1 min). Then,KOH/MeOH solution (2N; 0.4 ml) was added andthe sample was mixed again with the Ultra-turrax


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(max velocity for 1 min). After centrifugation FAMEwere recovered in the upper phase.

2.5.2 Two stepsTotal intramuscular lipids were extracted from 1 gof fresh muscle (5 replicates) adding 60 ml of asolution of chloroform/methanol (2:1) and omoge-nizing for 3 minutes, according to the method ofFOL [4] modified by Boselli et al. in 2001 [10]. Thesample was warmed for 20 min at 60°C, then 60 mlof a solution of chloroform/methanol (2:1) wasadded and another omogenization was carried out.After filtering with a vacuum pump, 34 ml of a KCl(0,88%) solution was added to the sample, and itwas left at -20°C overnight. The organic phase wasrecovered and dehydrated with anhydrous sodiumsulphate for 2 hours at the same temperature. Theorganic phase was filtered and the solvent wasremoved with a rotary evaporator. The fat wasrecovered in 2 ml of hexane and 20 μl of aKOH/MeOH solution (2N) was added in order toproduce fatty acid methyl esters for the CG analy-sis. After mixing (1 min), the sample was centrifu-gated to separate fatty acid methyl esters fromsoaps, according to the method already describedin a previous paper [3].

2.6 ANALYTICAL CONDITIONSFatty acid methyl esters were separated using aTRACETM GC Ultra (Thermo Electron Corporation,Rodano, Milano, Italy) equipped with the Ultra FastModule (UFM), a Fast Flame Ionization Detector(FFID) and a UFM-Carbowax column, 5-m long, 0.1mm i.d, 0.2 μm film thickness. 1 μl of FAME extract was analyzed in an ultra fastmode, with a temperature program ranging from150°C (held 10 sec) to 240°C (90 sec, 102°C/min)and held at 240°C for 2.30 minutes.The injection (PVT split mode) was operated with asplit ratio 1:150 and constant flow operating modeat 0.5 ml/min (helium as carrier gas). The injectorand detector temperature were both set at 240°C.Fatty acid peaks were recorded and integratedusing a Chrom-Card software (ver. 2.3.3, ThermoElectron Corporation, Rodano, Milano, Italy) andidentified by comparing retention times with FAMEstock solution.

2.7 STATISTICAL ANALYSISQuantification of FAME was based on the internalstandard technique and peak areas were correct-ed using FID response factors (RF) for each fattyacid, estimated by a previous calibration proce-dure with standards.Data are presented as means of five replicates, asconcentration (mg of fatty acid per g of fresh mus-cle). Coefficients of variation (RSD) were calculat-ed as SD x 100/mean.

Differences between methods were analyzed byone-way ANOVA and principal component analysis(PCA). When a significant effect (p < 0.05) wasdetected, the comparative analyses were conduct-ed using a post hoc Tukey test (STATISTICA, ver. 8,Stat. Soft Inc., Tulsa, OK, USA).

3. RESULTS AND DISCUSSION

3.1 ENZYMATIC DIGESTIONIn order to depress the matrix effect of meat, a pre-ventive enzymatic protein hydrolysis was carriedout. Among the different proteolityc enzymes,some requisites were considered as fundamental:optimal pH, affordability, rate of reaction, anddegree of liquefaction. Because of this, pepsine,which is characterized by a very high rate of reac-tion, was excluded due to its low pHopt = 1-2.Under these conditions, polyunsaturated FA(PUFA) can be damaged, while a partial triglyc-eride hydrolysis could be very likely as well, with anincrease of free FA. The inability of KOH/MeOH inderivatizing these FA would increase yield loss.Many other enzymes, such as trypsin and chy-motrypsin, were too expensive, or their action wastoo specific. Eventually, papain met all the definedprerequisites and it has a pHopt ≅ 6, similar to meat.

3.2. FA COMPOSITIONFA meat composition (Table I) showed perfectcomparable results between the FOL and FD test-ed methods, both for high (> 10%) and for low FAconcentrations (< 10%). On the contrary the enzy-matic method tend to underestimate FA both forhigh (> 10%) and for low FA concentrations (< 10%). Only for FA with the FD procedure < 1%some differences could be detected. In particular,C20:4 showed the greatest discrepancies. The roleof this FA in membrane lipids can explain the bestextraction with the FOL method. With its carefulsmashing of the tissue due to the kind of solventused, it should be more efficient in extracting thesekinds of lipids.RSDs of the absolute concentrations of FA (Table I)were below 10% in FD samples, a very remarkablevalue, considering they were the expression of thewhole process; on the contrary RSDs were higherthan 10% for the FA values resulting from the FOLmethod. Saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA) are better extract-ed with the FD method compared to the FOL one.PUFA seems to be better extracted with the FOLmethod. The C20:4 in FD samples are the only fattyacids that have a higher variability in terms of RSDs %.Although the preventive digestion of the samplewas thought as a means to eliminate the complexprotein framework of the matrix, it was a double-edged weapon. On drying, in fact, the proteic


hydrolizate generates a glassy mass difficult to bepenetrated by the MeOH/KOH/hexane mixture. Theobvious result was a depletion of the quantitativeyield, as shown in Table I. For these reasons thepre-digestion of the sample was rejected. On theother hand on adding the KOH/MeOH to FD sam-ple, dried meat was transformed into a gummymatter that was not easy to homogenize, as well.The use of a little spatula to detach the samplefrom the wall of the test tube is essential to reachrepeatability as good as those reported in Table I.The omission of this step affected yield as well.Tukey test (Table I) sum up these observations.FOL and FD gave comparable FA amounts. In fact,only C18:1, C20:1, and C20:4 were statistically dif-ferent. In particular, FOL seemed to underestimateC18:1 and C20:1, while C20:4 was higher with thisprocedure.In all cases, however, the discrepancies are tinyexcept for C20:4, one of the most sensitive FA todegradation.The other two methods were perfectly comparable,with freeze-drying apparently slightly more efficient(but not statistically different). Furthermore, theRSD values of the freeze-drying method, whenconsidering the absolute quantities, turned out tobe lower (in some cases even tenfold lower) thanthose of the other two methods, pointing out the

excellent repeatability of this procedure.The application of PCA to the sample set (Figure 1)confirmed and explained the considerations so farexpressed. FD was the most repeatable methodand it was very similar to the FOL one. In fact, theywere at the same height on PC1, which explained

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the vast majority of the total variability (86.19%). FDsamples have scores in a range of values between-4 and -2 and FOL ones between -4 and 2. AllFAME gave a contribution on PC1 in a very compa-rable way, with a negligible effect on PC2. Themain differences were concentrated on PC2, evenif it explains the 8.69% of the total variability. Themain contributor to the variation on PC2 is due to

some kinds of PUFA; in fact, only C20:4 and C18:3n-6 have an important effect on PC2, but an oppo-site sign (Figure 2). FOL samples are separated onPC2 because of the weight of C20:4 values vari-ability, considering the positive scores that charac-terize FOL cluster. C18:3 n-6 values seem to deter-mine the cluster position of FD samples.Digested samples were the most dispersed andthe most different from the standard method. Therejection of this new proposed method was thusfurther justified. A typical chromatogram of FAME inpork samples is represented in Figure 3.

3.3. TIME OF ANALYSISFrom a simple comparison of analysis time, freeze-drying seems to be not competitive. In fact, waterremoval is longer than the FOL extraction. Howev-er, this latter method requires a continuous pres-ence of an operator, while the FD required only alimited amount of time for sample freezing and toset up the freeze-drier. Consequently the wholeprocess will not require any operator support. Inaddition, it can work up a large number of samplesat the same time. In our laboratory a small 4-portfreeze-drier could theoretically operate at least 80samples simultaneously. Moreover, no lipid losscan occur. In spite of no experimental trial beingcarried out to support this hypothesis, it is veryunlikely that lipids can vaporize, even at the very


low pressure involved. Finally, by eliminating a realextraction step the use of expensive solvents andthe related problem of their disposal are limited.Comparing the two proposed methods and theFOL one (Table II), the freeze-drying of the freshsample followed by direct base-catalyzed trans-methylation, and the same procedure with a pre-ventive enzymatic digestion of the sample, only theformer gave good results. As a matter of fact, theFD was even better in terms of cost and ease ofautomation making this method particularly suit-able for a large number of sample processing.Nevertheless the method worked very well, andthese important details will be studied in the forth-coming research. At the moment, we are workingon samples with a different fat content and we areimproving the dispersion of the sample inKOH/MeOH solution, which is of crucial impor-tance for reaction yield. Moreover, the problem ofPUFA is a problem still open. Operating undermilder conditions of time and temperature wouldenhance the yield for these substances.Nevertheless the method worked very well, andthese important details will be studied in the forth-coming research.

4. CONCLUSIONS

Freeze-dried meat samples before the transmethy-lation step with KOH/MeOH seems to be a goodalternative to the FOL method followed by thederivatisation. This latter method is very labour-intensive, time and solvent consuming. At themoment, further data of experiments carried out ona larger number of samples with different lipid con-tent are very encouraging.

ACKNOWLEDGEMENTS

The research was supported by a financial contri-bution from G.A.L. Antico Frignano and AppenninoReggiano.

REFERENCES

[1] C.M. Murrieta, B.W. Hess, D.C. Rule. Compar-ison of acidic and alkaline catalysts for prepa-ration of fatty acid methyl esters from ovinemuscle with emphasis on conjugated linoleicacid. Meat Sci 65, 523-529, (2003).

[2] P. Magni, R. Facchetti, D. Cavagnino, S. Tre-stianu. KNL05 In: Proceedings of 25th Interna-tional Symposium of Capillary Chromatogra-phy. Sandra. P., Riva del Garda, Italy (2002).

[3] A. Ficarra, D.P. Lo Fiego, G. Minelli, A. Anto-nelli. Ultra fast analysis of subcutaneous porkfat. Food Chem 121, 809-814, (2010).

[4] J. Folch, M. Lees, G.H. Sloane Stanley. A sim-ple method for the isolation and purification oftotal lipides from animal tissues. J. of Biol.Chem. 226, 497-509, (1957).

[5] E.G. Bligh, W.J. Dyer. A rapid method of totallipid extraction and purification. Can J. ofPhysiol and Pharm 37, 911-917, (1959).

[6] A.I. Carrapiso, C. García. Development inlipid analysis: some new extraction tech-niques and in situ transesterification. Lipids35, 1167-1177, (2000).

[7] R. Eggers, U. Sievers, W. Stein. High pressureextraction of oil seed. J. Am. Oil Chem. Soc.62, 1222-1230, (1985).

[8] M.D. David, J.N. Seiber. Comparison ofextraction techniques, including supercriticalfluid, high-pressure solvent, and Soxhlet, fororganophosphorus hydraulic fluids from soil.Anal. Chem. 68, 3038-3044, (1996).

[9] L.S. Conte, O. Leoni, S. Palmieri, P. Capella,G. Lercker. Half-seed analysis: rapid chro-matographic determination of the main fattyacids of sunflower seed. Plant Breeding 102,158-165, (1989).

[10] E. Boselli, V. Velazco, M.F. Caboni, G. Lercker.Pressurized liquid extraction of lipids for thedetermination of oxysterols in egg-containingfood. J. Chromatography A., Elsevier, 239-244, (2001).

Received July 28, 2011Accepted January 24, 2012

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The seeds of Aleppo pine (Pinus halepensis Mill.) cultivated in Jordan were evaluated fortheir proximate composition and fatty acid, sterols, and amino acid contents. The fat, pro-tein, ash and total carbohydrate contents (on dry matter basis) were 32.1, 29.8, 5.5 and32.6%, respectively. The major fatty acids were linoleic (55.4%) and oleic (23%) acids.The seed oil content of Δ-5-olefinic acids was 8.7%. The unsaturated/saturated fatty acidsratio was 10.4. The major phytosterols were β-sitosterol (75.4%), campesterol (11.4%)and Δ-5-avenasterol (9.8%). The amino acid profile is characterized by high glutamic acidand arginine contents (193.7 and 140.3 mg/g protein respectively) and very low ratio oflysine to arginine (0.13). In conclusion, the high content of protein and unsaturated fattyacids, and the high ratio of unsaturated to saturated fatty acids, as well as the presence ofbioactive components including sterols, Δ-5-olefinic acids, and the low ratio oflysine/arginine in Aleppo pine seeds contribute to a healthy heart diet. Keywords: Fat, Fatty Acids, Sterols, Δ-5-Olefinic Acids, Protein, Amino Acids

Semi e composizione dell’olio di semi di Pino d'Aleppo (Pinus halepensisMill.) cresciuto in GiordaniaI semi di pino d'Aleppo (Pinus halepensis Mill.) coltivati in Giordania sono stati valutatiper la loro composizione in acidi grassi, steroli e il contenuto di aminoacidi.Il grasso, il contenuto totale di carboidrati, proteine e ceneri (sulla sostanza secca) sonostati 32,1, 29,8, 5,5 e 32,6%, rispettivamente. Gli acidi grassi più importanti sono l’acido linoleico (55,4%) e l’acido oleico (23%) l’olioè caratterizzato dal possedere acidi Δ-5-olefinici il cui contenuto è 8,7%. Il rapporto di acidi grassi insaturi/saturi è stato del 10,4. I fitosteroli principali sono il β-sitosterolo (75,4%), campesterolo (11,4%) e Δ-5-avenasterolo (9,8%). Il profilo aminoa-cidico è caratterizzato da un elevato contenuto di acido glutammico e arginina (193,7 e140,3 mg/g di proteine, rispettivamente) e il rapporto molto basso di lisina/arginina(0,13). In conclusione, l'elevato contenuto di proteine e di acidi grassi insaturi e l'elevato rappor-to di acidi grassi insaturi e saturi, così come la presenza di componenti bioattivi tra cuisteroli, acidi Δ-5-olefinici, e il basso rapporto di lisina/arginina nei pinoli Aleppo contri-buiscono ad una dieta corretta. Parole chiave: Grassi, Acidi grassi, Steroli, Acidi Δ-5-olefinici, Proteine, Aminoacidi

Seeds and seed oil compositions ofAleppo Pine (Pinus halepensis Mill.)

grown in Jordan

S.K. Tukan1*K. Al-Ismail1

R.Y. Ajo2

M.M. Al-Dabbas1

1Department of Nutrition andFood Technology, Faculty of

Agriculture, The University of Jordan,

Amman, Jordan

2Applied Science Department, Al-Huson University College

*CORRESPONDING [email protected]

Tel: +962 6 5355000Fax: +962 6 5527287


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1. INTRODUCTION

Aleppo pine (Pinus halepensis Mill.) is a large conif-erous evergreen tree. It is naturally distributed in theMediterranean, extending from Morocco in the westto Jordan in the east and from France in the north toPalestine in the south [1] and it covers an area ofabout 3.5 million Ha [2]. It is native to Jordan, andconsidered the dominant pine species in the naturalpine forests. It usually grows naturally on high alti-tudes, mostly over 700 m. Pine forests in Jordancover a total area estimated to be 13000 Ha compris-ing of about 30% of the total forest area (43000 Ha)in the country. The Aleppo pine tree is widespread inJerash, Ajloun, Dibbin and the Zay forest areas whichfall in the sub-humid bioclimatic zone, the Mediter-ranean type climate with a mild sub-humid winter anda long xerothermic period. It is also found in parks,sidewalks and also in gardens of old houses. It growson a variety of soils and tolerant to drought and heat,and it is widely used in reforestation [3, 4]. According to Jordanian seed collectors, who areusually women, cones of the Aleppo pine are usual-ly picked up from trees closed and freshly maturedwhen the color changes from green to bright brown,the typical color of freshly matured cones. The conesare kept until scales separate and seeds arereleased (usually between 8-10 weeks). The seedsare edible and have been traditionally used for foodin many countries of the Mediterranean [5]. In Jordanand Palestine, the seeds are eaten as a snack eitherraw or roasted; they are also used in several tradition-al foods; they are a significant component of malbanfruit leather prepared from semolina-thickened con-centrated grape juice, mainly produced in the Pales-tinian town of Hebron. The seeds give a crunchytaste to the chewy fruit leather. The seeds are alsoadded to porridge made from whole wheat, andused in liberal amounts in the production of tradition-al popular crunchy yeast-leavened bread called quz-mat prepared from whole wheat flour, olive oil andvarious oil seeds. Pine seeds are important for the local population inJordan especially to women; they can improve theirincome; enhanced seed consumption can contributeto family food security and better nutrition in additionto their possible health benefits. Furthermore, there iscurrently increased interest in underutilized plantsglobally for their multiple uses as an important dietarycomponent or as functional foods, in addition to theirrole in environmental sustainability [6]. Some information is available about Aleppo pineseeds grown in Europe and North Africa (northernand western regions of the Mediterranean) but stud-ies on seeds grown in the south eastern region of theMediterranean are scarce. The aim of this study,therefore, is to investigate the nutritive value of seedsgrown in Jordan (Mediterranean sub-humid biocli-

matic zone) with respect to proximate composition,lipids and amino acids composition.

2. MATERIALS AND METHODS

SAMPLE COLLECTION AND PREPARATIONAbout 2 kg of Aleppo pine seeds were purchasedfrom the local market. The seeds were ground usinga Krups coffee grinder. The ground seeds weresieved using 1-2 mm screen and stored in polyethyl-ene bags at freezing temperature until analysis.

PROXIMATE ANALYSIS Moisture, fat, protein and ash of the seeds weredetermined according to the standard AOAC meth-ods (1990) [7]. Crude protein was calculated byusing nitrogen conversion factor of 5.3 [8].

LIPID EXTRACTIONAbout 100 g of the ground seeds were immersed in500 ml hexane in a dark flask and stirred using mag-netic stirrer for 7 hrs at room temperature. The mix-ture was centrifuged for 10 min at 200 rpm. Thesupernatant was filtered using Whattman no.2 filterpaper. The extraction process was repeated once.The collected solvent was removed using rotaryevaporator at 40°C. The fat obtained was placed in aclosed bottle (50 ml) at freezing temperature untilanalysis.

FATTY ACID COMPOSITIONFatty acid methyl esters (FAMEs) of the oil extractedfrom the seeds were prepared according to EC Reg-ulation no. 2568/91 method [9]. Briefly, 50 mg of lipidextract was weighed, dissolved in 2 ml hexane (GCgrade) and mixed by vortex for 1 min. A 200 μl of 2M-potassium hydroxide prepared in anhydrousmethanol was added and mixed for 30 sec. until thesolution became clear, and then 200 μl of acetic acidwas added and mixed for 30 sec. The preparedmethyl esters were analyzed using capillary GLC col-umn (Restek, Rtx-225, USA, cross-bond 50%-cyanopropylmethyl 50%-phenylmethyl polysiloxane,60 m, 0.25 mm/D, 0.25 μm df) immediately afteresterification by injecting 1μl of the hexane layerthrough the injection port of the GLC (model GC-2010, Shimadzu. Inc., Koyoto, Japan). The FAMEswere injected after adjusting the GLC conditions; col-umn oven temperature was 165°C for 10 min,increased to 185°C 1°C/min and kept at 185°C for 1min, then increased to 220°C 3°C/min and kept at220°C for 20 min. Injector temperature was 240°C,flame ionization detector temperature was 260°C,flow rate 0.8 ml/min Helium and split ratio used was80. FAMEs were identified using chromatogram offatty acids standard (12-24 carbon atoms). The Δ-5-olefinic acids were identified by comparison of theirequivalent chain lengths (ECLs) with those of authen-


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er S433 (Sykam GmbH, Eresing, Germany) accord-ing to standard AOAC methods (1990) [7]. Briefly, 10ml of 6N HCl were added to 0.1 g ground seedssample in a test tube. The sample was hydrolyzed at110°C for 20 hours under continuous stirring thenallowed to cool to room temperature. The hydrolyzatewas transferred quantitatively with distilled water, fil-tered through filter paper to remove sediments, andthen evaporated to dryness under vacuum at 65°C.The hydrolyzate was dissolved in 1ml buffer (pH 2.2).A known volume (100 μl) was injected (done by anautosampler at temperature 12°C) into the aminoacid analyzer to determine the amino acid profile.Tryptophan content was then determined using thebasic hydrolysis method [11]. The ground seeds (0.1g) were treated with 4.2 M sodium hydroxide (10 ml)and hydrolyzed at 110°C for 20 hours. After cooling,the pH was adjusted to 4.5, and the mixture wasmade up to a certain volume. A programmable col-umn oven with temperatures ranging from 37-74°Cwas fitted as standard on the analyzer, a specialreagent pump “ninhydrin pump” was used to deliverninhydrin with a flow rate of 0.25 ml/min, and “bufferpump” with a flow rate of 0.45 ml/min was used. Theinert gas (N2) was supplied with a pressure of 0.5bars to prevent buffer/reagent oxidation and contam-ination. Dual channel photometer for the amino aciddetection was done at λ = 440 nm for proline, OH-proline and tryptophan, and at λ=570 for all otheramino acids.

3. RESULTS AND DISCUSSION

CHEMICAL COMPOSITIONThe chemical composition of Aleppo pine seeds isgiven in Table I. The seeds are a good source of oil(32.1% DM). When compared with seeds grown inother regions, the oil content of seeds in this study washigher than Turkish pines (21.1%) [12] but slightlylower than those obtained from Australia (34.1%)where seeds are used as aviary food [1] and fromFrance (35%) [10], but the value was much lower thanthe values obtained from Tunisian pine seeds (43.3%and 48.12 DM) [5, 13] respectively. Variation in com-position might be due to differences in the degree ofmaturation of seeds and cone age as well as variationsin environmental and ecological conditions.

tic related standards. ECLs were determinedaccording to Wolff and Bayard [10] with 16:0, 18:0,18:1, 18:2, 18:3, 20:0, 22:0, 22:1, 24:0 fatty acidsmethyl esters as reference components (Supelco,USA).

STEROL AND SQUALENE ANALYSESThe analysis of sterols in seed oil was carried outaccording to the method described by EC 2568/91[9]. For oil saponification, 5 g of the oil was weighedinto a 50 ml volumetric, 500 μg of squalane and 500μg α-cholestanol as internal standards and 50 ml of2 M KOH in methanol were added to the oil sample,the oil was heated under reflux in a water bath at70°C for about 1 h. The unsaponifiables wereextracted three times with 80, 70 and 60 ml diethylether. The pooled extract was then washed severaltimes with 50 ml water until the washing solutionbecame colorless with phenolphthalein. The solventwas removed under low pressure using a vacuumrotary evaporator. A solution of 10% w/v of theunsaponifiables in chloroform was prepared and 50μl of this solution was applied on a silica gel TLCplate (20×20 cm) for sterol isolation using n-hexa-ne/ethyl ether 60/40 (v/v). The plate was sprayed withethanolic solution of 2,7-dichloroflurosceint sodiumsalt (0.2% w/v). Sterols band was identified under UVlamp at 254 nm by comparing the Rf value with thatof β-sterol. The sterols were scrapped and extractedthree times with 3 ml chloroform. The solvent wasevaporated and the sterol-TMS derivatives were pre-pared using a solution composed of 5 volumes pyri-dine, 2 volumes of hexamethyldisilazan and 1 vol-ume of trimethylchlorosilan at 40°C for 20 min, andthen TMS solution was evaporated using extra purenitrogen gas. The derivatized TMS sterols wereredissolved in 1 ml hexane (GC grade) and mixed for30 seconds in a vortex mixer then centrifuged for 5min. The derivatized sterols were analyzed usingcapillary GC column (Restek, USA, crossbond 5%-diphenyl 95%-dimethyl polysiloxane, 30 m, 0.25mm/D, 0.1 μm df) immediately after trimethylsilylationby injection 1 μl of the hexane layer through the injec-tion port of the GLC. The GLC conditions used were;column oven temperature was 255°C for 40 min,injector temperature was 270°C, Detector tempera-ture was 280°C, flow rate was 1ml/min He, and splitratio used was 80:1. For squalene analysis theremaining unsaponifiables were derivitized as men-tioned for sterols and analyzed by GC. The GLC con-ditions used were; column oven temperature was200°C for 2 min, increased to 300°C 2°C/min andkept at 300°C for 12 min. the other GC conditionswere as those mentioned for sterols.

ANALYSIS OF AMINO ACIDS The amino acids profile of Aleppo pine seeds wasobtained using Sykam automatic amino acid analyz-


On the other hand, protein content (29.8%) of Alep-po pine seeds in this study is higher than that foundby other researchers. Values obtained for seedsgrown in Tunisia [5] and Australia [1], were 22.7 and24.8% respectively. The high protein content in thisstudy could be due to increased exposure of pineseeds in Jordan to high temperatures and waterstress which were reported to increase protein con-tent in other types of seeds [14]. The seeds also con-tain a good amount of ash (5.3%) but lower than thevalues for Tunisian (8.3%) [5] and Australian (7.7%)seeds [1].

FATTY ACID COMPOSITION Fatty acid composition of oil extracted from Jordan-ian Aleppo pine seeds is shown in Table II. As withother pine species [15], linoleic acid in this study isalso the predominant fatty acid (56.1%), followed byoleic (23.37%), palmitic (4.15%), sciadonic (3.82%),pinolenic (3.80%) and stearic (3.28%) acids. TheTable also shows linolenic acid is a minor constituent(0.66%) of pine seed oil. Unsaturated fatty acidsaccount for 90.2% of total fatty acids, with linoleicand oleic acids comprising 79.5% of the total unsat-urated fatty acids; the health benefits of these acidsare well documented. Diets rich in monounsaturatedfatty acids are recommended to prevent cardiovas-

90


cular diseases; they decrease low density lipoprotein(LDL)-cholesterol levels when replacing saturatedfatty acids [16, 17]. Saturated fatty acids in Jordanian pine seeds consti-tute to less than 10% of total fatty acids. These resultsagree with those reported by Wolff et al. [15], andNasri et al. [13] for oil extracted from seeds of theAleppo pine grown in France and Tunisia respective-ly, but differ from the results of Cheikh-Rouhou et al.[5] who reported much higher contents of total satu-rated fatty acids (16.8%) as well as relatively highpalmitic acid (8.7%) for Tunisian seed oil, that havecontributed to the high total fat content. Furthermore, the ratio of polyunsaturated to saturat-ed fatty acids in this study is very high (10.4:1), it ishigher than that obtained by Wolff et al. [15], Nasriand Tirki [18] and Cheikh-Rouhou et al. [5]; theirratios were (9.3:1; 8.87:1 and 4.9:1 respectively).With respect to Δ-5-olefinic acids, Jordanian Aleppopine seed oil, based on the ECLs determination, hadthe fatty acids: 18:2 cis-5, 9; 18:3 cis 5, 9, 12; and20:3 cis -5, 11, 14 at concentrations 1.0, 3.8, and3.82%, respectively (Table II). Total content of Δ-5-olefinic acids was 8.65% as compared to 9.1% forseeds cultivated in France [15] and 7.84% for thosecultivated in Tunisia [18]. These acids are known tobe specific to Pinus species and were used as a tax-onomic marker, and their content varies between 0.6- 31% [19]. Pinolenic acid (18:3cis -5, 9, 12) in thisstudy comprises about 44% of total olefinic acids. Itis also the main Δ-5-olefinic acid in various pinespecies, and its content varies between 0.16 - 14.9%of total fat [20]. Furthermore, the Δ-5-olefinic acids may have healthbenefits, they were found to decrease serum triglyc-eride levels in rats [21], while some studies onhumans using pine seed oil rich in pinolenic acid,suggest they may act as a natural appetite suppres-sant in overweight women [22].

UNSAPONIFIABLE CONTENTThe unsaponifiable lipid fraction of plant-based foodscomprises of the bioactive components phytosterolsand squalene. The unsaponifiable content of thestudied seed oil was 1.68% of total fat (Table III). Thisvalue is slightly lower than that found for TunisianAleppo pine [5] but relatively higher than the valuesobtained for oil from other pine species [18]. Theunsaponifiable content of seed oil in this study iscomparable to those of vegetable oils such as cornoil, palm oil and cottonseed oil [23].

STEROLS COMPOSITION AND SQUALENE CONTENT Phytosterols content of Aleppo pine seeds oil in thisstudy (Table III) was very high (6453 mg/kg oil), itaccounted for 38.4% of the unsaponifiables. Thesterol content of this seed oil is higher than the con-tent of some vegetable oils such as peanut (900-


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Phytosterols decrease blood levels of total choles-terol and low density lipoprotein (LDL) cholesterol, arisk factor in coronary heart disease, through com-petitive inhibition of cholesterol absorption [28, 29,30]. The National Cholesterol Education/Adult Treat-ment III program guidelines recommend intake ofplant sterols/stanols as part of a heart healthy eatingplan [30], while the Food and Drug Administrationapproved the health claim regarding their role inreducing risk of heart disease. Although their absorp-tion (≤ 5%) is low compared to cholesterol (35-70%),small amounts of phytosterols can be found in circu-lation and may influence other physiological func-tions. They were reported to possess anti-cancer,anti-inflammatory, anti-atherogenicity, and anti-oxida-tion activities [29, 31] Squalene is specific for vegetable oils, particularlyolive oil. The squalene range in olive oil is 136-709mg/kg oil, while its range in the other vegetable oilsis between 2-332 mg/ kg oil [32]. The squalene con-tent in the seed oils of our study was 144 mg/kg oil,which is similar to most vegetable oils. Squalene is a precursor in the synthesis of choles-terol and steroid hormones. It has antioxidant poten-tial and found to reduce lipid peroxidation in the reti-na. Although no direct relationship between squa-lene consumption and incidence of cancer has beeninvestigated, its high content in olive oil was suggest-ed to be a protective factor that might explain the lowincidence of certain cancers in the Mediterraneanpopulation [33].

AMINO ACID COMPOSITION Previous studies on pine seeds as food for humanswere mostly done on lipid fractions, including bioac-tive components, but data on amino acids content isvery limited. The amino acid composition of Aleppopine seeds is shown in Table IV. The major aminoacids are glutamic acid (5.5% dry weight) and argi-nine (4.0%) comprising of about one third (33%) ofseed protein. Storage proteins are usually rich inariginine, the amino acid with the highest nitrogencontent [34, 35]. Arginine is believed to have ahypotensive effect on humans; this is due to itsmetabolism to nitric oxide a potent vasodilator [36,37]. Furthermore, the lysine to arginine ratio (0.13:1)in this study is very low, it is similar to macadamianuts but lower than many other nuts which rangefrom 0.19 for hazel nuts to 0.57 for pistachios [38,39],and was 0.67 for sesame globulins [40]. The low ratioof lysine to arginine has been indicated a health ben-efit, it is believed to lower the risk of hypercholes-terolemia and atherosclerosis in laboratory animals[40, 41]. Furthermore, aspartic acid, tryptophan andhydroxyproline occur in pine seeds in significantamounts (2.43, 2.30 and 2.23% DM respectively).Table IV also shows that the contents of essentialamino acids in this study were much lower than those

2900 mg/kg), soybean (1800-4500 mg/kg), and sun-flower (2400-5000 mg/kg) oils, but lower than thoseof corn (7000-22100 mg/kg) and sesame (4500-19000 mg/kg) oils [23]. Data in Table III indicate that β-sitosterol is the mostabundant sterol (75.3% of total sterols) followed bycampesterol (11.4%), similar results were obtainedby Cheich-Rouho et al. [24] for Tunisian oil fromseeds of the same species and by Nasri et al. [13] forseed oil from other species of pine. Both sterols arethe most commonly occurring phytosterols in plantfoods [25]. In addition, Δ-5-avenasterol (9.8%) ispresent in significant amounts in Jordanian pine seedoil but was not reported or detected in Tunisian seedoil [24]. Its content in oils of seeds from other pinespecies studied [13] and many other seed oils wasmuch lower than the values obtained in this studyexcept for peanut (8.3-18.8) and coconut oils (26-29%). On the other hand, Δ-7-avenasterol and Δ-5,24-stigmastadienol are present in very low amounts(0.89% and 0.81% respectively). The seed contentsof sterols, sitosterol, campesterol, Δ-5-avenasteroland stigmasterol were 198, 23, 19 and 0.04mg/100g, respectively. The pine seed content ofsitosterol is higher than those of many grains,legumes and seeds except chickpeas and peas;campesterol content was also higher, except forpeas and sesame seeds, whereas stigmasterol wasmuch lower [26].Cholesterol content in the Jordanian Aleppo pine seedsis very low (0.4% of total sterols). Cholesterol is specif-ic to animal fat, and occurs at low levels in most of veg-etable fats and oils ranging from not detected up toabout 4%, except in tomato seed oil which containsabout 20% of total sterols [27]. Cholesterol in plants isa precursor of steroidal saponins and alkaloids.


reported for Australian pine seeds [1] except formethionine which was higher in Jordanian seeds.Methionine is known to have positive effect on zincabsorption [42].

CONCLUSIONS

The results of the study show that Aleppo pine seedsare an important dietary component, they are calorieand protein dense food and have components withimportant biological functions, thus contributing tofighting hunger, maintenance of a good and healthyheart and to disease prevention. Accordingly, con-sumption of seeds as part of a healthy diet andlifestyle is recommended especially by the localcommunities.

ACKNOWLEDGEMENT

The author S. K. Tukan would like to thank the Dean-ship of Scientific Research at the University of Jordanfor the financial support of this work.

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[40] T. Rajamohan, P.A. Kurup. Lysine: arginineration of a protein influences cholesterol metab-olism part-1 studies on sesame protein havinglow lysine: arginine ratio. Indian Journal ofExperimental Biology 35, 1218-1223 (1997).

[41] D. Kritchevsky. Dietary Protein and Experimen-tal Atherosclerosis. Annals of the New YorkAcademy of Sciences 180-187 (1997)

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Received, September 5, 2011Accepted, October 28, 2011

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Candida rugosa lipase was immobilized by entrapment in alginate beads. The optimiza-tion of the immobilized lipase preparation in regard to water content was performed. Theobtained immobilized biocatalyst preparation was tested in acidolysis of rapeseed oil witha lauric acid (9:1 w/w) process. The activity of Candida rugosa lipase was high andincreased with the increasing of the processing time. The immobilized Candida rugosalipase preparation was used as the biocatalyst in interesterification of goose fat and rape-seed oil mixture (2:3 w/w). The starting mixture and the interesterified products were sep-arated by column chromatography into a pure triacylglycerol fraction and non-triacylglyc-erol fraction. It was found that the concentrations of free fatty acids and partial acylglyc-erols increased after interesterification. The sn-2 and sn-1,3 distributions of fatty acids inthe TAG fractions after interesterification indicated that Candida rugosa lipase kept itspositional nonspecificity after immobilization. The interesterified fats had reduced oxida-tive stability, as assessed by Rancimat induction times and peroxide values.Keywords: Candida rugosa lipase, immobilization, enzymatic interesterification, goosefat, rapeseed oil

Interesterificazione di grasso d'oca e miscela di olio di colza utilizzandolipasi di Candida rugosa immobilizzata in perline di alginatoLa lipasi di Candida rugosa è stata immobilizzata tramite intrappolamento in perline dialginato. È stata eseguita l’ottimizzazione rispetto al contenuto d'acqua della preparazionedi lipasi immobilizzata. Il preparato biocatalizzatore immobilizzato ottenuto è stato testatocon un processo di acidolisi di olio di colza con acido laurico (9:1 w/w). L'attività lipasi-ca della Candida rugosa era alta e cresceva con l'aumentare del tempo di elaborazione. La preparazione di lipasi immobilizzata di Candida rugosa è stata utilizzata come biocata-lizzatore nella interesterificazione della miscela di grasso d'oca e olio di colza (2:3 w/w).La miscela di partenza e i prodotti interesterificati sono stati separati mediante cromato-grafia su colonna in una frazione pura di triacilgliceroli e una frazione non triacilgliceroli-ca. Si è riscontrato che le concentrazioni degli acidi grassi liberi e degli acilgliceroli par-ziali aumentavano dopo l’interesterificazione. Le distribuzioni sn-2 e sn-1,3 degli acidigrassi nelle frazioni TAG dopo l’interesterificazione erano casuali. I grassi interesterificatiavevano una stabilità ossidativa ridotta, come stimato dai tempi di induzione Rancimat edai valori di perossido.Parole chiave: Lipasi di Candida rugosa, immobilizzazione, interesterificazione enzima-tica, grasso d'oca, olio di colza

Interesterification of goose fatand rapeseed oil mixture using

Candida rugosa lipase immobilizedin alginate beads

K. Tarnowska1*M. Kostecka2

A. Piotrkowicz3

M. Łobacz1

B. Kowalski1

1University of Life Sciences Faculty Of Food Sciences

Warsaw, Poland

2PhD Student at Faculty of FoodSciences, University of Life

Sciences (SGGW), Warsaw, Poland

3M.Sc. Student at Faculty of FoodSciences, University of Life

Sciences (SGGW), Warsaw, Poland

*CORRESPONDENCE: Katarzyna Tarnowska

University of Life Sciences (SGGW)Faculty of Food SciencesDepartment of Chemistry

Laboratory of Food Chemistry159C Nowoursynowska Street

02-787 Warsaw, PolandFax: +48 22 5937625

e-mail:boleslaw@[email protected]

[email protected]


INTRODUCTION

Poultry fat, one of the by-products in the meatindustry, is an inexpensive raw material producedin considerable quantities. Despite of its produc-tion and ready availability as a coproduct of chick-en or goose meat production, poultry fat, unlikebeef tallow, is usually not used separately in otherfood or nonfood applications [1]. Animal fats, ingeneral, are of dietary concern because of their rel-atively high long-chain (C16 and C18 carbonatoms) saturated fatty acids (SFA) content. Com-pared to the fat of mammals, poultry fat is regard-ed to be relatively safe for consumers. It is charac-terized by a highly desirable composition of fattyacids. Chicken fat contains about 60% unsaturatedfatty acids, while goose fat 70% (70.83% in abdom-inal fat and almost 73% in subcutaneous fat) andoleic acid is dominating [2]. Both chicken and goose fat are characterized byrelatively good polyunsaturated/saturated(PUFA/SFA) acids ratio ranging from 0.3 to 0.8 andhigher proportions of PUFA with at least two unsat-urated sites, but it reduced oxidative stability ofthese fats [1, 2, 3]. Poultry fat has not been the focus of many investi-gation till XXI century [4]. There is a potential optionof technologically modifying this fat by blending it,fractioning, or by its interesterification.One of the possible ways of animal fats modifica-tion is their interesterification with vegetable oils.The selection of vegetable oil depends on theregional production and purpose of fat modifica-tion. In Poland rapeseed oil can be taken intoaccount, because rapeseed is cultivated on alarge scale. Compared to the other vegetable oilsoccurring on the polish market, rapeseed oil con-tains low levels of saturated fatty acids (about 7%).A high content of oleic acid (about 60%) andantioxidants ensure its relatively good oxidativestability. There are two types of interesterification present-ly in use: chemical and enzymatic. Depending onthe catalyst used and operating conditions of theprocess, products with different structures of tria-cylglycerols can be obtained. Enzymatic inter-esterification relies on the use of random or

regiospecific and fatty acid-specific lipases ascatalysts, whereas for chemical interesterification,metal alcoholate catalysts are typically employed[5]. The application of lipases in the oil and fatindustry has several advantages over the chemi-cal method, including the above mentioned selec-tivity and positional specificity against fatty acidsexchange, milder reaction conditions, toleranceto water and free fatty acids, reduced environ-mental pollution, availability of lipases from a widerange of sources and the ability to improve lipas-es by genetic engineering. The use of lipases andother enzymes has been limited due to their con-siderably unstable nature and the resultingrequirement of stringent conditions, such as aparticular pH and temperature. In chemical andbiochemical reactions, purified enzymes can berather costly and to discard them after each use isnot economical. Retrieval of enzymes from thereaction medium, however, can be difficult andtherefore researchers have explored the immobi-lization of enzymes. Immobilized enzymes typical-ly exibit greater stability and enzyme activity overa broader range of pH and temperature [6, 7]. Microencapsulation of lipases has proved to be asuccessful immobilization technique. Microcap-sules can be easily separated from the reactionmedium by filtration, and immobilized catalystshave the potential of being used in several reactionbatches [7, 8, 9]. The objective of this study was to immobilize Can-dida rugosa lipase by entrapment in alginatebeads to use it as the biocatalyst in interesterifica-tion of goose fat and rapeseed oil mixture. Theactivity of the catalyst used (immobilized lipase)and its dependence on the water content in thepreparations were also studied. The selectedchemical and physical properties of the interester-ified fats were compared with those obtained fromfat blends before interesterification. The productscan be used as "low trans" margarine oils or com-ponents of frying fat.

MATERIALS AND METHODS

MATERIALSThe extra virgin/first pressing rapeseed oil (RSO)

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(ZT "Kruszwica" S.A., a Bunge Company, Poland)was commercially refined, bleached and deodor-ized. Goose fat (GF) (abdominal fat) was obtainedfrom the local market and it was refined andbleached in a laboratory. The mixture 2:3 (w/w)GF:RSO was prepared by weight. The parametersof the goose fat, rapeseed oil and goose fat: rape-seed oil mixture are listed in Table I. The main fattyacid composition and distribution among sn-2 andsn-1,3 positions of triacylglycerols in goose fat andthe initial blend are shown in Tables II and III.

CATALYSTAs the catalyst, lipase from Candida rugosa TypeVII (Sigma, USA) was used. The lipase has areported activity of ≥ 700 unit/mg (One unit willhydrolyze 1.0 microequivalent of fatty acid from a

triacylglycerol in 1 h at pH 7.2 at 37°C).

CHEMICALS AND SOLVENTSAll chemicals and solvents used in this study werereagent grade and their producers are mentionedthroughout the paper.

METHODS Immobilization of lipase (the entrapment in alginatebeads)10 ml enzyme solution contained 250 mg lipasewas mixed with 10 ml of 4% sodium alginate(Aldrich, USA). The mixture was slowly droppedinto 100 ml 0.2 M CaCl2 using a pasteur pipette.This resulted in polymerization of alginate, whichtook approximately 0.5 h to be completed. Gel beads with entrapped enzymes were collected

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by filtration through a sieve.

Optimization of the immobilized lipase preparation in regardto water contentGel beads with entrapped lipase were dried in37°C in variously variants:- to constant mass and then watered on the level

of 10% in regard to lipase preparation weight,- to constant mass and then watered on the level

of 60% in regard to lipase preparation weight,- to 50% water content in regard to lipase prepara-

tion weight,- to 20% water content in regard to lipase prepara-

tion weight,- to 10% water content in regard to lipase prepara-

tion weight.Dried as specified enzyme preparations were usedas a catalyst for the interesterification of goose fatand rapeseed oil mixture (2:3 w/w; 40°C, 2 h). Afterinteresterification process the acid value wasmeasured.

Estimation of the activity of the immobilized lipase prepara-tionThe determination of the activity of the immobilizedlipase was done based on the model acidolysisreaction using lauric acid (Merck, Germany) andrapeseed oil mixture. There is no lauric acid in tria-cylglycerols of rapeseed oil, therefore the percent-age lauric acid content incorporated during theacidolysis process into triacylglycerols of rapeseedoil could be an estimation of lipase transesterifica-tion activity.Directly before the acidolysis rapeseed oil (27 g)and the lauric acid (3 g) mixture was thermallyequilibrated at 45°C in stoppered flasks positionedin a thermostated mineral oil bath shaker. Afterthermal equilibration of the sample immobilizedlipase preparation was added. The catalyst prepa-ration mass was fitted so that the protein contentwas 0.23 g. The acidolysis was carried out withcontinuous shaking for 2, 4, 8, 21, 24 and 48 hours.Experiments performed with a catalyst contained20% of water. This water content was a result ofdrying lipase preparation after the immobilizationprocess. Filtering off the catalyst stopped the aci-dolysis. The fat after acidolysis was deacidifiedwith ethanolic KOH solution, washed with water toremove soaps and dried. The remained fractions offat: triacylglycerols, mono- and diacylglycerols(TAG, MAG + DAG) were analysed by GLC.

Interesterification of goose fat and rapeseed oil (2:3 w/w/)mixtureFlasks contained goose fat and rapeseed oil mix-ture (30 g) were positioned in a thermostated min-eral oil bath shaker. After thermal equilibration(40°C) immobilized lipase preparation was added.

The catalyst preparation mass was fitted so that theprotein content was 0.23 g. The water content inthe biocatalyst was established after immobiliza-tion on the 20% level in regard to the lipase prepa-ration weight. The interesterifications were carriedout with continuous shaking for 1, 2, 3, 4 and for 8hours. Interesterification was stopped by filteringoff the biocatalyst.

DETERMINATIONS Determination of protein content in the catalystIn gel beads with entrapped lipase the nitrogenpercentage content were measured using CHNSanalyzer model Vario EL III (Elementar, USA). Thenthe results were recalculated into the protein con-tent.

Determination of the acid valueAcid values (AV, in mg KOH/g) were determined bytitration of fat samples dissolved in a mixture ofethanol and diethyl ether 1:1 (v/v) with 0.1 M potas-sium hydroxide water solution using phenolphtalein(1% in ethanol) as an indicator. Analyses were car-ried out in accordance with the Standard PN-ENISO 660:2005 and PN-EN ISO 660:2005/Ap1:2007.

Determination of free fatty acidsFree fatty acids (FFA, in %) content was calculatedbased on acid values and the values of averagemolar masses of fatty acids obtained from GCresults. The average molar masses of fatty acidsdisplayed: MGF = 274 g/mol, MRSO = 278 g/mol,MGF:RSO (2:3) = 276 g/mol (before interesterifica-tion) and MGF:RSO (2:3) = 276 g/mol (after interester-ification).

Determination of peroxide value Fat samples were dissolved in the mixture of chlo-roform: acetic acid = 2:3 (vol/vol) and 1ml solutionof saturated potassium iodide was added. Flaskswith fat were closed and kept in darkness. After 5minutes several drops of 2% starch solution wereadded. Fat samples were titrated with 0.001 Msodium thiosulfate solution (Polish Standard PN-ISO 3960:1996).

Separation of fat samples into triacylglycerol and non-tria-cylglycerol fractionFats before and after interesterification were sepa-rated into triacylglycerols (TAG) and non TAG frac-tion, refered to as polar fraction (PF), by columnchromatography on silica gel (SG 60, 70-230mesh, Merck, Germany). The TAG were eluted withthe mixture of petroleum ether:diethyl ether = 87:13vol/vol and the PF fraction contained FFA, monoa-cylglycerols (MAG) and diacyglycerols (DAG) waseluted with ethyl ether. The weight percentages ofTAG and PF were determined after the evaporation

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of the eluting solvent. Analyses were carried out inaccordance with the Standard PN-EN ISO8420:2004 and PN-EN ISO 8420:2004/AC:2008.

Positional distribution of fatty acids among sn-2 and sn-1,3positions of TAGThe fatty acids composition in the triacylglycerolsamples obtained from the mixtures before andafter the enzymatic and chemical interesterifica-tions was determined based on the results of gaschromatography (GC) analysis. Samples were pre-pared and analyzed based on the Standards PN-EN ISO 5509:2001 and PN-EN ISO 5508:1996.The positional distribution of fatty acids among thesn-2 and sn-1,3 position of triacylglycerols wasdetermined using the method developed by Brock-erhoff (selective hydrolysis of TAG) [10-12]. TheBrockerhoff method employs the ability of the pan-creatic lipase enzyme to selectively hydrolyze esterbonds in the sn-1,3-position of the TAG molecules.The hydrolysis products were separated by thinlayer chromatography (TLC), and the compositionof fatty acids in the isolated sn-2-monoacylglyc-erols was examined by GLC. Next, the compositionof fatty acids in the sn-1,3-position of TAG wascomputed from the results obtained and the knowl-edge of the composition of fatty acids in the triacyl-glycerols. The participation of particular fatty acidsin the sn-2-position of TAG was also determined.

APPARATUS AND PROCEDURESGC measurements of fatty acid methyl estersThe methyl esters were analyzed on a ShimadzuGC17A gas chromatograph equipped with a flameionization detector and a BPX capillary column(30m × 0.22mm × 0.25μm film thickness). The tem-perature program was: 60°C for 1 min, after whichthe temperature was increased at a rate of10°C/min to 170°C and from 170 to 230°C at a rateof 3°C/min. The temperature was kept at 230°C foranother 15 minutes. The injection and detectortemperature were 225 and 250°C, respectively.Nitrogen was used as a carrier gas, flowing at arate of 1 mL/min. The composition of fatty acidswas expressed as weight percentages.

Oxidative stability of fats studied by Rancimat measure-mentsA Metrohm Rancimat model 679 (Herisau, Switzer-land) was used for the measurement of oxidationstability indexes. The test was carried out twice foreach sample, using 2.5 g of deacidified oil/fat. Allmeasurements were taken at 100°C. The tempera-ture of the conductivity tube was kept at a constant21°C and the air flow rates were set at 20L/h foreach experiment. The Rancimat induction times(τRancimat, in hours), supplied automatically by theapparatus control software had a precision of two

decimal places for results between 0 and 10 hoursand one decimal place for results over 10 hours.The results reported in this study are the average ofthe two measurements taken for each sample.

Statistical evaluation of the experimental dataStatistica 8.0 (StatSoft Inc., USA) package wasused for statistical evaluation of experimental data.

RESULTS AND DISCUSSION

OPTIMIZATION OF THE IMMOBILIZED LIPASE PREPARA-TION IN REGARD TO WATER CONTENTThe water content is a very important factor in sys-tems where lipase as a catalyst is used. To favorsynthesis, the water content should be kept as lowas possible. However, water plays an importantrole in the stabilization of the three dimensionalstructure of proteins. Additionally, lipases are acti-vated only when adsorbed in an oil/water interface,so a certain necessary amount of water for lipaseactivity must be guaranteed. With this in mind, thefirst task of this study was to optimize the immobi-lized lipase preparation in regard to water content.As seen in Table IV the result of the drying of lipaseimmobilized preparation to constant mass andthen watering to 10 and 60% was a lack in activityof this lipase. In these cases the changes in acidvalues were slight if compared with the initial mix-ture (0.22 mg KOH/g). It indicated that hydrolysisdid not occur. Water was inaccessible for the lipaseboth when it was added to fat and to lipase prepa-ration. Probably, drying caused excessive rigidityof the lipase molecule, so it was impossible to getthe right dimensional structure for the catalyticfunction. A similar situation was observed for dry-ing to 10% water content in regard to the lipasepreparation weight. The acid value after the inter-esterification process increased, however, basedon our earlier experiments with Lipozyme RM IMand Novozym 435 watered to 10% [13, 14], we canassume that activity of Candida rugosa lipase islow. Whereas the water content in the catalystadjusted to 50% caused a shift in the balancebetween hydrolysis and synthesis in the directionof hydrolysis. In this study optimum water contentin biocatalyst preparation appeared to be 20% andthis level of water was kept in further interesterifica-tion processes.

ACTIVITY OF IMMOBILIZED LIPASE The significant question of this investigation was toverify if entrapped in alginate beads Candidarugosa lipase is active in the interesterificationprocess. The determination of the transesterifica-tion activity of the immobilized lipase was donebased on the model acidolysis reaction performed


using lauric acid and rapeseed oil mixture. There isno lauric acid among other fatty acids of rapeseedoil triacylglycerols. Accordingly it is obvious thatthe effectiveness of acidolysis with lauric acid canbe monitored by the determination of this acidincorporation into the triacylglycerols of rapeseedoil. The products of acidolysis contained TAG,DAG, MAG, FFA from rapeseed oil and unreactedlauric acid. As the fatty acids were removed bydeacidification the final products after acidolysisconsisted of TAG and DAG + MAG fractions. Theresults of experiments (acidolysis) presented inFigure 1 are time dependent. The activity of Candi-da rugosa lipase (the yields of the incorporation offatty acid into acyloglycerols) during acidolysisreactions increased with the increasing of acidoly-sis time. The incorporation of lauric acid was in therange of 1 to almost 8%. The obtained biocatalystactivity was recognized as rather high if comparedto our other experiments concerning enzymaticacidolysis of beef tallow with lauric acid catalysedby commercial immobilized lipase preparations[13].

INTERESTERIFICATION OF GOOSE FAT AND RAPESEEDOIL (2:3 w/w) MIXTUREThe results of determinations and analyses ofgoose fat, rapeseed oil and their 2:3 GF:RSO blendbefore interesterification are listed in Table I. TheGF, RSO and their non-interesterified mixture dis-played: 0.08 – 0.17% of FFA, 0.38 – 10.80% of DAG+ MAG and 89.03 – 99.54% TAG. The τRancimat

values were between 2.75h to 13.95h. As expect-ed, the addition of rapeseed oil resulted in anincrease in content of linoleic and α-linolenic acidsand a decrease in content of palmitic and stearicacids, compared to raw poultry fat (Tables II andIII). The addition of the RSO oil also caused a sig-nificant increase in oxidative stability of the mixture,due to compensation of the lack of antioxidants inGF [15].It is well-known that during the interesterification offats/oils two reactions take place simultaneously:hydrolysis and resynthesis of triacylglycerols. As aresult, interesterification products contain, in addi-tion to TAG, certain amounts of free fatty acids, dia-cylglycerols and monoacylglycerols, that constitutethe polar fraction (PF). The hydrolysis reactionreduces the final yield of the interesterified TAG butaccelerates the interesterification reactions by rais-ing the concentration of DAG, which are essentialintermediates.The interesterified products contain the larger per-centage of PF in comparison to the initial mixture asdepicted in Table V. The FFA content is in the rangeof 6.68% to 30.51% depending on the reactiontime. The increase of FFA content was proportionalto the time of reaction. The acid value (AV) is relat-ed with the degree of hydrolysis reaction and canbe the measure of the FFA content in a final fattyproduct. The direction of changes of AV deter-mined for fats after interesterification is the same asfor FFA percentages. The contents of partial acyl-glycerols in post-reaction mixtures increased when

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compared with the starting blend and reached amaximum level of 18.59% for the mixture after 2hours of interesterification. Generally, the percent-age of MAG in the interesterified product is quitelow (about 1%) because MAG are interesterifiedvery fast as discussed by Mohamed et al. [16] andby Ledóchowska et al. [17].As the concentrations of polar components duringthe modification process increased, the efficiencyof TAG in the post-reaction mixtures decreased.The decrease of nonpolar fractions was inverselyproportional against the time of process. Theamounts of TAG and polar fraction in the interester-ified samples strongly depend on the water contentin the system. Ledóchowska et al. [17] andTarnowska et al. [18] showed that the higher theamount of water in the reaction system, the higherthe FFA and MAG + DAG contents and the smallerthe TAG content. The preparation of immobilizedCandida rugosa lipase contained 20% water and ithas a relatively high moisture degree if comparedto the commercial immobilized lipase preparation(Lipozyme RM IM and Novozym 435) used in ourearlier experiments [18]. In those studies the watercontent above 10% caused predominance hydrol-ysis over TAG resynthesis. The differences in opti-mum water content, required for high interesterifi-cation activity, may be caused by various immobi-lization methods and a kind of support. Enzymaticinteresterification reaction systems must contain acertain amount of water, because the reactiontakes place on the water/oil boundary phase. Thiswater has two functions: it is required by theenzyme to maintain an active hydrated state and itis a reactant for producing partial acylglycerolsand FFA from TAG by hydrolysis [5]. The presenceof these by-products has detrimental effects on thequality of the resulting fats, for example on oxida-tive stability [19].The oxidative stability of obtained modified prod-ucts was also measured and the results are shownin Table V. The peroxide values (PV) determined foresterified fats increased if compared with the start-

ing mixture. The maximum PV value (mmol O2/kg)was measured for the mixture after 8 hours of inter-esterification. This fat product contained the highestpolar fraction content, which exhibits prooxidativeproperties. The Rancimat tests performed for inter-esterified fats (Table V) showed that their τRancimat

times were drastically reduced (4.7-2.6 h) com-pared with 13.45 h for the starting blend. It may becaused by esterification of antioxidants from rape-seed oil (esters have no antioxidant properties) ortheir deactivation during the transesterificationprocess. As mentioned earlier, the oxidative stabil-ity could be influenced by the high FFA, DAG andMAG contents. The product of interesterificationperformed for 4 hours and proved to be the mostresistant to oxidation compared to the other inter-esterified fats. Such a decrease of thermal-oxida-tive stability after enzymatic interesterification wasobserved also in our earlier experiments [14, 18]and by other authors in their studies concerninginteresterification of beef tallow with vegetable oils[17]. Modified lipids (structured lipids – SLs) differ fromnaturally occurring lipids/fats because they containfatty acids in quantities, proportions or positions inwhich they do not exist naturally. The main fattyacid composition of triacylglycerols in the initialblend and TAG isolated from products after variousreaction times are shown in Table VI. It can be seenthat the composition of fatty acids in TAG of the ini-tial mixture and of the products of interesterificationdiffer significantly in some analyzed cases. Theinteresterified lipids (TAG) contain mainly oleic(over 60%), linoleic (~ 14%), palmitic (12-13%) andα-linoleic (5-6%) acids. After a comprehensive sta-tistical analysis of the obtained results the influenceof the reaction time on the TAG composition wasobserved. It was found that interesterification had asignificant influence on the contents of the C16:1-cis, C18:2 all-cis and C18:3 all-cis acids, and asmaller influence on C16:0, C18:0 and α-C18:1-cisacids when compared to the initial mixture. Statisti-cally significant changes in C16:0 and C18:0 acids


contents were observed for reactions with pro-longed interesterification times. When SLs are synthesized from animal fats for fooduses, it is often desirable to reduce the saturatedLCFA content of the starting fat and increase theMUFA and PUFA content as a means of improvingthe nutritional quality of the targeted structuredlipid [1]. We have observed higher MUFA andPUFA contents for TAG isolated from the productsof reaction compared to goose fat. The MUFA andPUFA contents for the interesterified blend were62.8-63.1% and 18.3-20.5%, respectively. Thesecontents were much higher than those measuredfor raw goose fat (MUFA – 60.3%, PUFA – 8.7%). Itis worth noting, that after interesterification the con-tent of trans C18:1 isomer retained on the same

level as for initial blend.The properties of fats depend not only on the fattyacids composition of triacylglycerols but on theirstructure as well. Fatty acyl groups located at thesn-2 position of TAG are regarded as being easierto absorb by the human digestive system than sim-ilar fatty acyl groups located at the sn-1,3 positionsof TAG [3]. Hydrolysis using pancreatic lipase,which is regioselective to positions 1-3, leadsmainly to the formation of free fatty acids originat-ing from outer TAG positions, as well as 2-monoa-cylglycerols. The 2-monoacylglycerols can easilypass through the intestinal wall, in contrast to freefatty acids, and especially their saturated forms,which readily form insoluble calcium salts prevent-ing their absorption. Hence, saturated fatty acids

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occupying this position will be highly absorbed. Itwas therefore important to investigate the structureof goose fat: rapeseed oil (2:3 w/w) mixture beforeand after interesterification. The major fatty acidscompositions and distributions among sn-2 andsn-1,3 positions for nonesterified goose fat, its ini-tial mixture with rapeseed oil and TAGs isolatedfrom selected products are listed in Tables II, IIIand VII-IX. Triacylglycerols isolated from goose fatcontained mainly oleic acid in the inner sn-2 posi-tion, whereas palmitic, palmitooleic and stearicacids were located mostly in the outer sn-1,3 posi-tions. Participation of C14:0, C18:2 all-cis andC18:3 all-cis acids in the sn-2 position was 33.3-36.7% and their distribution was random. The addi-

tion of rapeseed oil to goose fat caused somechanges in TAG structure. Palmitic, palmitooleicand stearic acids occupied mostly the sn-1,3 posi-tions, but distribution of oleic acid was statistical.The PUFA (C18:2 all-cis and C18:3 all-cis) werelocated in the sn-2 position.The native Candida rugosa lipase is positional non-specific [5], however after immobilization its prop-erties could change. The obtained data (Table VII,VIII) suggests that some positional randomizationof fatty acids in TAG, after interesterification lasted1 and 4 hours, has occurred. On the other hand,the sn-2 percentage data showed that althoughthere is some degree of randomization, the distri-bution of some fatty acids is far from statistical

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(33.3%). The "disturbance" of structure of TAG of fatmodified in interesterification lasted 8 hours mayresult from acyl migration in the TAG species dur-ing prolonged interesterification time and highwater content in the reaction system [20]. Acylmigration is caused mainly by the existence of par-tial acylglycerols, especially diacylglycerols(DAG), necessary and unavoidable intermediatesin lipase-catalyzed interesterification reactions.Acyl migrations between positions 2 and 1(3)occur via identical processes. The 1,2(2,3)-DAG'sare not thermodynamically stable and tend tospontaneously transform into 1,3-DAG. There aremany factors possibly influencing acyl migration:acyl donor (different migration rates for differentfatty acids), water content, solvents, the catalyzingenzyme and its support, temperature, reaction time[20,21].

CONCLUSIONS

The studies performed in this work can be con-cluded as follows:• immobilization Candida rugosa lipase by entrap-

ment in alginate beads makes the biocatalystpreparation active in acidolysis rapeseed oil withlauric acid and in interesterification of goose fatand rapeseed oil mixture

• the optimal water content in immobilized lipasepreparation in these reaction systems appearedto be 20% in regard to the mass of the lipasepreparation

• Candida rugosa lipase kept its positional non-specificity after immobilization

• interesterification of GF+RSO (2:3 w/w) blendproduces new fats which do not have naturalequivalents; these fats, when purified, are suit-able for use in various food applications, thuswidening the utilization of goose fat

ACKNOWLEDGEMENTS

The authors are grateful to Prof. Ewa Białecka -Florjańczyk and Dr. Jolanta Krzyczkowska fromthe Department of Chemistry University of Life Sci-ences (SGGW) in Warsaw for aiding in the creationof the immobilization formula.

LITERATURE

[1] K.T. Lee, T.A. Foglia, Fractionation of chickenfat triacylglycerols: synthesis of structuredlipids with immobilized lipases. J. Food Sci.65 (5), 826-831 (2000).

[2] B. Biesiada - Drzazga, Description of select-ed characteristics of muscle and fat tissue of10-week white Koluda W31 geese. Acta Sci.Pol., Technol. Aliment. 5 (2), 47-54 (2006).

[3] K.T. Lee, T.A. Foglia, Synthesis, purificationand characterization of structured lipids pro-duced from chicken fat. J. Am. Oil Chem. Soc.77 (10), 1027-1034 (2000).

[4] M. Kostecka, M. Łobacz, Lipids from chickenfat - invaluable (underestimated) fat. Part I -Chicken fat characteristic and chosen modifi-cation methods. Technol. Progress in foodprocessing, 1, 98-103 (2009) [in Polish].

[5] A.G. Marangoni, D. Rousseau, Engineeringtriacyloglycerols: The role of interesterifica-tion. Trends in Food Sci. & Technol. 6, 329-335(1995).

[6] KE Pye, Present and future trends in enzymetechnology and its application. (Ed.) OlsonAC, Cooney CL, Immobilized enzymes in foodand microbial processes. Plenum Press, NewYork 1-17 (1974).

[7] Z.D. Knežević, S.S. Šiler-Marinković, L.V.Mojović, Immobilized lipases as practical cat-alysts. APTEFF I-208, 151-164 (2004).

[8] S.S. Betigeri, S.H. Neau, Immobilization oflipase using hydrophilic polymers in the formof hydrogel beads. Biomaterials 23, 3627-3636 (2002).

[9] M. Matsumoto, K. Ohashi, Effect of immobi-lization on thermostability of lipase from Can-dida rugosa. Biochem. Eng. J. 14 (1), 75-77(2003).

[10] H. Brockerhoff, A stereospecific analysis oftriglycerides. J. Lipids Research 6, 10-15(1965).

[11] H. Brockerhoff, Stereospecific analysis oftriglycerides. Lipids. 6, 942-956 (1971).

[12] B. Drozdowski, Wpływ budowy glicerydów iwystępujących w nich kwasów tłuszczowychna mechanizm hydrolizy enzymatycznej(Effect of the structure of acylglycerols and itsfatty acids on enzymatic hydrolysis mecha-nism). Scientific Books of the Gdansk Univer-sity of Technology Chemistry, 25, 3-86 (1974)(in Polish).

[13] B. Kowalski, K. Tarnowska, E. Gruczyńska,Enzymatic acidolysis of beef tallow with lauricacid. Riv. Ital. Sostanze Grasse, 284-289(2004).

[14] B. Kowalski, K. Tarnowska, E. Gruczyńska, W.Bekas, Chemical and enzymatic interesterifi-cation of beef tallow and rapeseed oil equal-weight blend. Eur. J. Lipid Sci. Technol. 106,655-664 (2004).

[15] A. Raemy, I. Froelicher, J. Loeliger, Oxidation oflipids studied by isothermal heat flux calorime-try. Thermochim Acta. 114, 159-164 (1987).

[16] H. M. A. Mohamed, S. Bloomer, K. Hammadi,Modification of fats by lipase interesterifica-tion I: Changes in glyceride structure. Fat Sci.Technol. 11, 428-431 (1993).

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[17] E. Ledóchowska, E. Wilczyńska, Comparisonof the oxidative stability of chemically andenzymatically interesterified fats. Fett/Lipid100, 343-348 (1998).

[18] K. Tarnowska, M. Kostecka, B. Kowalski, E.Gruczyńska, M. Kowalska, Improvement ofnutritional quality of low-value animal fats byinteresterification with rapeseed oil. In:Advances in research and technology ofrapeseed oil. Monograph – part II. Ed. E.Szłyk, Scientific Press UMK, Toruń, 109-128(2009).

[19] E. Ledóchowska, I. Datta, Wpływ frakcji nietri-acyloglicerolowej na stabilność oksydatywnątłuszczu przeestryfikowanego chemicznie ienzymatycznie (The influence of nonpolarfraction on the oxidative stability of chemical-

ly and enzymatically interesterified fats).Food. Science. Technology. Quality 18(1), 15-23 (1999) [in Polish].

[20] X. Xu, A.R.H. Skands, C.-E. Høy, H. Mu, S.Balchen, J. Adler-Nissen, Production of spe-cific-structured lipids by enzymatic interester-ification: elucidation of acyl migration byresponse surface design, J. Am. Oil Chem.Soc. 75, 1179-1186 (1998).

[21] X. Xu, S. Balchen, C.-E. Høy, J. Adler-Nissen,Pilot batch production of specific-structuredlipids by lipase-catalyzed interesterification: pre-liminary study on incorporation and acyl migra-tion. J. Am. Oil Chem. Soc. 75, 301-308 (1998).

Received, July 11, 2011Accepted, November 21, 2011

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A study was conducted to determine the suitability of six varieties of bambara groundnut(Vigna subterranea L. Verdc) seed oils, an under–utilized crop grown in Nasarawa State, Nige-ria. For this purpose, fatty acid composition of six different seed colours of bambara ground-nut was determined using standard analytical techniques. The most concentrated fatty acidswere oleic acid (17.54-18.49%) < palmitic acid (21.79-23.27%) < linoleic acid (34.04-35.62%). Arachidic and behenic acids were present in small quantities with none of themrecording up to 1.0% in any of the samples. Caprylic, palmitoleic, margaric, capric, lauric,myristic, arachidonic, erucic and lignoceric acids were all determined but not detected. Unsat-urated fatty acids predominated in all the samples with an adequate amount of essential fattyacid (linoleic and linolenic acids). Significant differences were observed (p < 0.05) in the fattyacid compositions among the bambara groundnut dehulled seed cultivars. The results ofphysicochemical properties of bambara groundnut varieties seed oils showed mean rangevalues of the following parameters: saponifications value (174.80-181.02 mg KOH/g); perox-ide value (9.10-11.05 meqO2/kg); iodine value (111.98-122.22 mg of I/100g); acid value(1.28-1.40 mg KOH/g); specific gravity at 25°C (0.874-0.881); unsaponifiable matter (2.39-2.47%); flash point (210-220°C); kinetic viscosity at 40°C (2.95-4.94). Generally, the valuesof the physicochemical parameters showed that the oils may be useful as edible oils due totheir stability as frying oils and may also be useful industrially for the manufacture of prod-ucts such as paints, liquid soaps and shampoos.Keywords: Vigna subterranea, seed oils, fatty acids, physicochemical parameters.

Valutazione delle caratteristiche chimico-fisiche e degli acidi grassi di seivarietà di olio di arachide della specie Bambara (Vigna subterranea L. Verdc) E’ stato svolto uno studio per determinare l'idoneità dell’olio di semi ricavato da sei varie-tà di arachidi bambara (Vigna subterranea L. Verdc), una coltura sottoutilizzata nello Statodi Nasarawa, in Nigeria. A tal fine è stata determinata la composizione degli acidi grassi,utilizzando le normali tecniche analitiche, di sei differenti semi, di colori diversi, di arachi-di bambara. Gli acidi grassi più rappresentati sono stati: l’acido oleico (17,54-18,49), <l'acido palmitico (21,79-23,27%), < l’acido linoleico (34,04-35,62%). Gli acidi arachicoe beenico erano presenti in piccole quantità con valori inferiori all’1,0% in tutti i campio-ni. Gli acidi caprilico, palmitoleico, margarico, caprico, laurico, miristico, arachidonico,erucico e lignocerico erano tutti determinati, ma non rilevati. Gli acidi grassi insaturi predominavano in tutti i campioni con adeguata quantità di acidi grassiessenziali (acidi linoleico e linolenico). Sono state osservate differenze significative (p < 0,05)nella composizione in acidi grassi tra le cultivar di semi di arachidi bambara decorticata. I risul-tati delle proprietà fisico-chimiche dell’olio di semi di arachidi della varietà bambara ha mostra-to valori medi gamma dei seguenti parametri: numero di saponificazione (174,80-181,02 mgKOH/g); di perossidi (9,10-11,05 meq/kg), iodio (111,98-122,22 mg di I/100g), numero di aci-dità (1,28-1,40 mg KOH/g), il peso specifico a 25°C (0,874-0,881); insaponificabile (2,39-2,47%), punto di infiammabilità (210-220°C); viscosità cinetica a 40°C ( 2,95-4,94). I parametri chimico fisici hanno dimostrato come i campioni esaminati possano essereutilizzati sia come alimento, sia come oli per frittura, naturalmente come molte sostanzegrasse possono essere utilizzate anche per la produzione di prodotti per l’industria cosme-tica e per quella delle vernici.Parole chiave: Vigna subterranea, oli di semi, acidi grassi, parametri fisico-chimici.

Evaluation of fatty acids andphysicochemical characteristics of

six varieties of bambaragroundnut (Vigna subterranea

L. Verdc) seed oils

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M.O. Aremu1*S. Mamman1

A. Olonisakin2

1Department of Chemistry, Nasarawa State University, PMB 1022, Keffi – Nigeria

2Department of Chemistry,Adekunle Ajasin University,

Akungba Akoko-Nigeria

*CORRESPONDING AUTHOR:e-mail: [email protected]


INTRODUCTION

Bambara groundnut (Vigna subterranea L. Verdc)species is of African origin and is widespread inthe south of the Sahara. It appears to have evolvedin the Savannas probably in West Africa, and itscompact habit seems to be an adoption to growingin hot and windy environments [1 – 2]. In terms ofproduction, bambara groundnut ranks third amongthe grain legume crops of the African lowland trop-ics after groundnut (Arachis hypogeae L.) andcowpea (Vigna unguiculata L. Walp) [3]. The crop is grown in the middle belt and northernregions of Nigeria. They are consumed fresh whensemi–ripe, as pulses when dry and mature orground to flour for the preparation of moi-moi (Yoru-ba tribe) or opa (Hausa tribe) [4]. No cultivars ofbambara groundnut have been named but geno-types are distinguished on the basis of seed attrib-utes such as colour, size and hardness and plantform (bushy or spreading). Sometimes names are based on the location wherethe seed was collected [5]. The colour of the seed varies from white to ivory,dark brown to red or black. There are various com-binations of these colours such as speckled, mot-tled, mosaic, variegated or patterned seeds with orwithout colouring around the eye [6]. However,bambara groundnut which was formally popular inthe northern part of Nigeria because it was wellfavoured by dry lands and environmental climaticconditions has been gradually replaced over thecourse of decades by the groundnut particularly inmore productive areas.

The bambara groundnut seed cannot be regardedas an oil seed [7 – 8] because of its low crude fatcontent but its oils are consumed as part of theseed. This work therefore reports on some fatty acid com-position and physicochemical parameters (saponi-fication value, proxide value, iodine value, acidvalue, specific gravity, unsaponifiable matter, flashpoint, kinetic viscosity) of bambara groundnutseed oils as part of our continuing research into

new vegetable fat and oil sources, to know whetherthe seed oils could be used for food and industrialapplications.

MATERIALS AND METHODS

COLLECTION AND TREATMENT OF SAMPLESMature and dry bambara groundnut (Vigna subter-ranea L. Verdc) seeds were collected from differentfarms located at Keffi, Garaku, Lafia, Akwanga,Wamba and Nasarawa towns in Nasarawa State,Nigeria. Six different bambara groundnut colourvarieties of cream with black eye, cream with blackspeck, cream, cream with violet spotted, lightbrown and dark brown, from respective towns wereidentified and labeled as VSK, VSG, VSL, VSA, VSW

and VSN (Table I). The various seeds were sortedand screened to remove the ones that were notmature. Boiling water was added to the seeds, leftovernight, and the seeds dehulled, dried in theoven at 45°C for 8 h. The dried seeds were finelyground into fine flour with a small sample mill [9],stored in polythene bags and kept in a refrigeratorat 4°C prior analyses.

EXTRACTION OF OILSOven dried sample was extracted in Soxhlet appa-ratus with redistilled n–hexane of Analar grade(British Drug Houses, London) for the recovery ofundiluted oil. The crude oil extract was made to befree of water by filtering through the anhydroussodium sulphate salt. The hexane was removedfrom the oil/hexane mixture by using a rotary evap-orator.

FATTY ACID ANALYSISThe oil extracted was converted to the methyl esterusing the method described by Akintayo andBayer [10]. About 2 mg crude oil sample wastransferred into a 5–10 ml glass vial and 1 ml of dia-zomethane ether solution added. The mixture wasshaken thoroughly and allowed to stand for 1 min.Then 16 μL of 3.33M CH3CONa/CH3OH solutionwas added; mixture shaken and allowed to standfor 10 mins after which 10 μL acetic acid was

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added. The equation of the transmethylation isshown below:

The fatty acid methyl esters were analyzed using aHP 6890 gas chromatograph powered with HPChemstation Rev. a 09.01 (1206) software fittedwith a flame ionization detector and a computingintegrator. Nitrogen was used as the carrier gas.The column initial temperature was 250°C rising at5°C/min to a final temperature of 310°C while theinjection port and the dector were maintained at310°C and 350°C, respectively. A polar (HP INNOWax) capillary column (30 m × 0.53 mm × 0.25 µm)was used to separate the esters. The peaks wereidentified by comparison with standard fatty acidmethyl esters obtained from Sigma Chemical Co.(St. Louis, MO, USA).

PHYSICO–CHEMICAL ANALYSESThe physico–chemical analyses of the oils forsaponification value, peroxide value, iodine value,acid value, specific gravity, unsaponifiable matter,flash point and kinetic viscosity were carried outaccording to the methods of AOAC [11].

STATISTICAL EVALUATIONThe statistical calculations included percentagevalue, grand mean, standard deviation and relativestandard deviation (RSD).

RESULTS AND DISCUSSION

The fatty acid composition of six varieties of bam-bara groundnut (Vigna subterranea) seed oils is

shown in Table II. The result showed that saturatedfatty acids present were palmitic (C16:0), stearicacid (C18:0), arachidic acid (20:0) and behenicacid (22:0). The monounsaturated fatty acids werepalmitoleic (C16:1), oleic (C18:1) while the polyun-saturated were linoleic (C18:2) and linolenic(C18:3). Linoleic acid had the highest concentra-tion of fatty acid with values ranging from 34.04%in cream to 35.62% in dark brown varieties. Theobservation in this report is in perfect agreementwith the earlier reports by some researchers [12 –19] that linoleic was the most concentrated fattyacid in legume seed oils. The second concentrat-ed fatty acid was palmitic acid with values rangingfrom 21.79% in cream to 23.27% in cream with vio-let varieties and average of 22.48±0.60% while thethird in position was oleic acid (17.54–18.49%). Ithas been reported that linoleic and oleic acids aremajor fatty acids in peanut, soya bean, chide pea,garden pea, broad bean and lentil while cowpea,black-eyed pea, kidney and California small whitebean have linoleic and linolenic acids as the majorfatty acids [15, 20]. Many lipids from legume seedscontain substantial amounts of saturated fattyacids, especially palmitic acid [21]. A higher pro-portion of either linoleic or linolenic acid is associ-ated with legumes containing insignificant amountsof lipids [22]. The arachidic (0.43–0.70%) andbehenic (0.20–0.82%) acids had the least values(Table II) with mean values of 0.58±0.11% and0.54±0.22%, respectively. The total lipid content oflegume values vary with variety, origin, locationand climate as well as seasonal and environmentalconditions, and the type of soil in which they aregrown [23]. Statistical analysis of the fatty acidcomposition in the present study showed that sig-nificant differences occurred (p < 0.05) among thefatty acid values of the samples. The only likely rea-son for this observation is due to genotypic differ-ences since all other information about the sam-ples are similar.

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The distribution of results in Table II into TSFA,MUFA, DUFA, TUFA, TUSFA, and oleic to linoleic(O/L) ratio is shown in Table III. TSFA ranged from33.37% in light brown to 35.32% cream with blackspeck varieties with an average of 34.45±0.77%and variation of 2.24%. These values are lowerthan TSFA mean value (54.51%) of dehulledAfrican yam bean [16] but higher than the valuesreported for the raw and processed pinto beans(9.0–12.9%) [19]. The total unsaturated fatty acids(TUSFA) which makes up ω-9 and ω-6 fatty acidsranged from 64.68 to 66.63%. The ω-6 and ω-3fatty acids have critical roles in the membranestructure [24 – 25] and as precursors ofeicosanoids, which are potent and highly reactivecompounds. Since they compete for the sameenzymes and have different biological roles, thebalance between the ω-6 and ω-3 fatty acids in thediet can be of considerable importance [20]. Therecommended ratio of linoleic to α–linolenic acidsin the diet should be between 5:1 and 10:1 [20].The ratios of linoleic to α–linolenic of six varieties ofbambara groundnut in the present study rangedbetween 4.0:1 to 4.3:1 with an average of 4.1:1.This shows that on a diet based mainly on onlydehulled varieties of bambara groundnut;FAO/WHO nutritional requirement will not be met.Saturated fatty acids and monosaturated fattyacids can be synthesized from carbohydrates andproteins. Also, positive protein balance can con-tribute to an overall energy balance in the sameway that positive carbohydrate balance does [20].A high fat, low–carbohydrate diet increases the riskfor onset of non–insulin–dependent diabetes melli-

tus (NIDDM) [26]. Thus, this may not occur in abambara groundnut diet.It has been concluded that relative to carbohy-drates, the saturated fatty acids elevate serumcholesterol, while the polyunsaturated fatty acidslower serum cholesterol [27 – 28]. The saturatedfatty acids (SFA); palmitic (C16:0), stearic (C18:0),arachidic (C20:0) and behenic (22:0) (Table II),have been established as the most important of thedietary risk factors in CHD [29]. There are threetypes of lipoproteins (protein–lipid complexes) inthe blood. These are low–density lipoproteins(LDL) in which 46% of molecule is cholesterol, etc.,high–density lipoproteins (HDL) which include 20%as cholesterol, and very–low–density lipoproteins(VLDL) which have 8% cholesterol. High levels oftotal blood cholesterol are associated with the inci-dence of CHD as well as high intakes of saturatedfatty acids [29]. A direct comparison of palmiticacid showed that it raises LDL cholesterol relativeto oleic acid, but that myristic acid was slightlymore powerful (but below the detection limit of GCin this work) [30]. The other major SFA in the bam-bara groundnut, stearic acid (C18:0), may not beas hyper-cholesterolemic as the other SFA (appar-ently because it is converted to oleic acid) [31]. The monounsaturated fatty acid (MUFA) of maininterest is oleic acid (C18:1) which is high in vari-eties of bambara groundnut (17.54–18.49%) (TableII) compared with varieties of African yam bean(0.16–1.81%) [16]. Polyunsaturated linoleic acidmoderately reduces serum cholesterol and LDLlevels [20]. Linoleic and α–linolenic acids are themost important essential fatty acids required for

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growth, physiological functions and body mainte-nance [22]. Bambara groundnut seed oils will par-ticipate well in these functions. The total essentialfatty acid (TEFA) values ranged from 42.81 to44.39% with an average value of 43.77±0.58% anda variation of 1.33% (Table III). The ratios of TUSFAto TSFA (P/S ratio) ranged from 1.83 in cream withblack speck to 2.0 in light brown varieties of bam-bara groundnut with an average of 1.90. This P/Sratio is better than the value of 0.52 reported for thekernel of Terminalia catappa (Tropical almond) [32]and that of varieties of African yam bean (0.54 to0.72) [16]. The relative amounts of PUFA and SFAin oils is important in nutrition and health. The ratioof polyunsaturated to saturated acids (P/S) ratio istherefore important in determining the detrimentaleffects of dietary fats. The higher the P/S ratio themore nutritionally useful is the oil. This is becausethe severity of artericosclerosis is closely associat-ed with the proportion of the total energy suppliedby saturated fats and polyunsaturated fats [33 –34]. The TEFA in dehulled varieties of bambaragroundnut are much higher than TEFA in rice,sorghum, millet and maize [35 – 36]. The bestsource of EFA (44.39%) among our samples is thedark brown dehulled variety (Table III). With the rel-atively high P/S mean ratio (1.90) for the bambaragroundnut seed oils and appreciable amount ofEFA, the arterogenic tendencies of this oil could beeasily suppressed [34]. The oleic/linoleic (O/L)acids ratio has been associated with high stability

and potentiality of the oil for deep frying fat [14].The O/L levels in the present study ranging from0.50 to 0.54 are lower than peanut oil (1.48) [14]hence bambara groundnut oils may be less stablecompared with peanut oil and may also not be use-ful as frying oil. The levels of RSD ranged from 0.96in TUSFA to 3.02 in MUFA.Table IV displays the physicochemical characteris-tics of bambara groundnut seed oils. The lightbrown to reddish – coloured oils had saponificationvalues ranging from 174.80 mg KOH/g in lightbrown to 181.02 mg KOH/g in cream with violetspotted. The values are higher than valuesobtained for some vegetable oils in the literature[10, 37 – 38]. This suggests that seed oils of bam-bara groundnut may be utilized in production of liq-uid soaps and shampoos. The peroxide and acidvalues ranged from 9.10–11.05 meq/kg and1.28–1.40 mg KOH/g, respectively. Peroxide valueis an indicator of deterioration of oils [10]. The per-oxide values in the present study are far below thevalue reported for Bilphia sapida seed oil whichwas only recommended for industrial use [10] butcomparable to melon seed oil [38]. Acid value isused as an indicator for edibility of an oil and suit-ability for use in the paint and soap industries [39].The low acid values of all the bambara groundnutseed oils indicate that they may be edible [37]. Theiodine values obtained for all the samples placethem in the drying group oil since drying oils havean iodine value above 100 [40]. The relative high

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iodine value may be an indication of the presenceof high unsaturated bonds and high susceptibilityto oxidative rancidity [41]. The specific gravity at25°C ranged from 0.874 in dark brown to 0.881 incream with black eye, indicating that the oil is lessdense than water and compared with cashew nutoil of 0.964 [42]. The unsaponifiable matter values(2.39–2.47%) are low and favourably comparedwith 2.44% reported for Malagasy legume seed oils[43]. High unsaponifiable matter content of fatsand oils have been reported to be an indication ofadulteration or contamination [42]. The flash points(°C) ranged from 210 in cream with black eye to220 in light brown while kinetic viscosity values at40°C were 2.95 to 4.94. The relative standard devi-ation (RSD) levels were relatively close rangingfrom 0.00 to 12.53 Table IV. This suggests thatgenotypic differences showed not much influenceon the physicochemical properties of bambaragroundnut seed oils.

CONCLUSION

The study had presented data on the concentra-tions of saturated and unsaturated fatty acids of sixvarieties of bambara groundnut (Vigna subterraneaL. Verdc.) grown in Nasarawa State, Nigeria. Somephysico–chemical properties of the seed oils werealso presented. There was a clear indication thatthe seed oils of all the six varieties contained a highlevel of polyunsaturatated fatty acids, making thema healthy low fat food. The suitability of the oils fordomestic use and utilization in the production ofpaints, liquid soap and shampoos are alsorevealed.

REFERENCES

[1] M.O. Aremu, O. Olaofe, C.A. Orjioke, Proxi-mate and amino acid composition of bambaragroundnut (Vigna subterranea), kersting’sgroundnut (Kerstingiella geocarpa) and scar-let runner bean (Phaseolus coccineus) proteinconcentrates. Riv. Ital. Sostanze Grasse 85,128–134 (2008).

[2] J.A. Duke, Handbook of Legumes of WorldEconomic Importance. Plenum Press, NewYork (1981).

[3] K.O. Rachie, P. Silvestre, Grain legumes. In:Food Crops of the Lowland, Tropics, Leaky,C.L. A and Will, J.B. (eds.). Oxford UniversityPress, Oxford, 41–74 (1977).

[4] M.O. Aremu, O. Olaofe, E.T. Akintayo, Mineraland amino acid composition of two varietiesof bambara groundnut (Vigna subterranea)and kersting’s groundnut (Kerstingiella geo-carpa) flours. Int. J. Chemstry 16(1), 24–30(2006).

[5] D.E. Kay, Food Legumes. TPI Crop Digest No3 Trop. Products Institute, London (1979).

[6] J. Coudert, Market openings in West Africa forcowpeas and bambara groundnut. Int. TradeForum 20(1), 14–15 (1984).

[7] M.O. Aremu, O. Olaofe, E.T. Akintayo, A com-parative study on the chemical amino acidcomposition of some Nigerian under–utilizedlegume flours. Pak. J. Nutrition 5(1), 34–38(2006).

[8] N.J. Enwere, Y.C. Hung, Some chemical andphysical properties of bambara groundnutseed and products. Int. J. Food Sci. and Nutri-tion 47(6), 469–475 (1996).

[9] M.O. Aremu, O. Olaofe, E.T. Akintayo, Nutri-tional qualities assessment of the presence ofhull in some Nigerian under–utilized legumeseeds. Bulletin of Pure and Applied Sciences24(1-2), 47–52 (2005).

[10] E.T. Akintayo, E. Bayer, Characterization andsome possible uses of Plukenetia conophoraand Adenopus brevilorus seeds and seedoils. Bioresource Tech. 85, 95–97 (2002).

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[12] A.I. Ihekoronye, P.O. Ngoddy, Integrated FoodScience and Technology for the Tropics Lon-don, Macmillan (1985).

[13] A.A. Paul, D.A.T. Southgate, McCance andWiddowsen’s “The Composition of Foods”,Royal Society of Chemistry, London (1985).

[14] W.D. Branch, T. Nakayama, M.S. Chennan,Fatty acid rariation among US runner typepeanut cultivars. J. Am. Oil Chem. Soc. 67,591–596 (1990).

[15] A. A. Oshodi, O. Olaofe, G.M. Hall, Aminoacid, fatty and mineral composition of pigeonpea (Cajanus cajan). Int. J. Food Nutri. 43,178–191 (1993).

[16] E.I. Adeyeye, A.A. Oshodi, K.O. Ipinmoroti,Fatty acid composition of six varieties ofdehulled African yam bean (Sphenostylisstenocarpa) flour. Int. J. Food Sci. and Nutri.50, 357–365 (1999).

[17] E.T. Akintayo, E. Bayer, Identification of oils byNMR spectroscopy. Riv. Ital. Sostanze Grasse79, 207–210 (2002).

[18] M.O. Aremu, O. Olaofe, E.T. Akintayo, L.Lajide, Characterization of some under–uti-lized legume seed oils by NMR spectroscopy.Oriental J. Chemistry 23(1), 81–87 (2007).

[19] S.S. Audu, M.O. Aremu, L. Lajide, Effect ofprocessing on fatty acid composition of pintobean (Phaseolus vulgaris L.) seeds. Int. J.Chem. Sci. 4(1), 114–119 (2011).

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Food and Nutrition Paper 57, Rome:WHO/FAO (1994).

[21] F.A. Lee, L.R. Mattick, Fatty acids of the lipidsof vegetables. I. Peas (Pisum satirum). J.Food Sci. 26, 273–275 (1961).

[22] D.K. Salunkhe, S.S. Kadam, J.K. Chavan,CRC Posthharvest Biotechnology of FoodLegumes. Boca Raton, FL: CRC Press (1985).

[23] R.E. Worthington, R.O. Hammons, J.R. Alli-son, Varietal differences and seasonal effectsof fatty acid composition and stability of oilfrom 282 peanut genotype. J. Agric FoodChem. 20: 727–730 (1972).

[24] D.V. Lynch, G.A. Thompson, Re–tailored lipidmolecular species: A tactical mechanism formodulating membrane properties. TrendBiochem. Sci. 9, 442–445 (1984).

[25] J.E. Kinsella, Possible mechanisms underly-ing the effects of n–3 polyunsaturated fattyacids, Omega–3, News, V: 1–5 (1990).

[26] J.A. Marshall, R.F. Hamman, J. Baxter, High-fat, low-carbohydrate diet and the etiology ofnon-insulin-dependent diabetes mellitus: TheSan Luis Valley diabetes Study. Am. J. Epi-demiol. 134(6), 590–603 (1991).

[27] A. Keys, J.T. Anderson, F. Grande, Predictionof serum cholesterol response of man tochanges in fats in the diet. Lancet 2, 959–966(1957).

[28] D.M. Hegested, L.M. Dusman, J.A. Johnson,G.E. Dallal, Dietary fat and serum lipids: Anevaluation of the experimental data. Am. J.Clin. Nutr. 57, 875–883 (1993).

[29] A. Bender, Meat and Meat Products in HumanNutrition in Developing countries, FAO Foodand Nutrition Paper 53. Rome: FAO (1992).

[30] P.L. Zock, J.H. De Vries, M.B. Katan, Impact ofmyristic and versus palmitic acid on serumlipid and lipoprotein levels in healthy womenand men. Arterioscier. Thromb. 14, 567–575(1994).

[31] A. Bonanome, S.M. Grundy, Effect of dietarystearic acid on plasma cholesterol andlipoprotein levels. New England J. Med. 318,1244–1248 (1988).

[32] A.M. Balogun, B.L. Fetuga, O. Osibanjo,Nutrient composition of the kernel of Termina-

lia catappa (Tropical almond). Nigerian J. Sci.24(1–2), 206–209 (1990).

[33] G. Honatra, Dietary fats and arterial thrombo-sis. Heamostasis 2, 21–52 (1974).

[34] A. Keys, Diet as a risk factor in cardiovasculardisease. Paper presented at the 148th Scientif-ic Meeting of the Nutrition Society, 6–7th July,1972. University of Surrey, Guildford (1972).

[35] A. Adeyeye, K. Ajewole, Chemical composi-tion and fatty acid profiles of cereals in Nige-ria. Food Chem. 44, 41–44 (1992).

[36] B. Holland, I.D. Union, H.D. Buss, Vegetables,Herbs and Spices, rth Supplement to McCaneand widdowson’s ‘The Composition of Foods’.London: Royal Society of Chemistry (1991).

[37] M.O. Aremu, O. Olaofe, E.T. Akintayo, Chemi-cal composition and physicochemical char-acteristics of two varieties of bambaragroundnut (Vigna subterranea) flours. J. Appl.Sci. 6(9), 1900–1903 (2006).

[38] M.O. Oladimeji, A.O. Adebayo, A.H. Adegbe-san, Physico–chemical properties of Hausamelon seed (Cucumeropsis edulis) flour. UltraScience 13(3), 374–377 (2001).

[39] I. E. Akubugwo, G. C. Chinyere, A. E.Ugbogu, Comparative studies on oils fromsome common plant seeds in Nigeria. Pak. J.Nutri. 7(4), 570–573 (2008).

[40] H.J. Jr. Duel, The Lipids: Their Chemistry andBiochemistry. Vol. 1 New York IntersciencePublishers (1951).

[41] O.U. Eka, Proximate composition of bushmango tree and some properties of dika fat.Nig. J. Nutr. Sci. 1, 33–36 (1980).

[42] M.O. Aremu, A. Olonisakin, D.A. Bako, P.C.Madu, Compositional studies and physico-chemical characteristics of cashew nut (Anar-cadium occidentale) flour. Pak. J. Nutr. 5(4),328–333 (2006).

[43] E.M. Gaydolel, J.P. Blanchimi, J.V. Rotoro-hovy, Triterpenealcohols, methysterols, sterolsand fatty acids in five Malagasy legume seedoils. J. Agric Food Chem. 31, 833–836 (1983).

Received, December 2, 2012Accepted, January 14, 2012

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Da diversi anni viene pubblicata una guida, a disposizione di chi lavora nel settore deilubrificanti, in cui sono riportati i controlli maggiormente utilizzati per la caratterizzazionedei prodotti petroliferi e lubrificanti e i relativi metodi di analisi pubblicati da Enti Nazio-nali ed Internazionali (UNI, CEI, ASTM, IP, ISO, IEC, EN).Quest’anno è stata fatta la revisione della tabella con un aggiornamento di tutti i metodipubblicati da gennaio a dicembre 2012.La struttura base della tabella non è stata modificata rispetto alla versione precedente:nella prima colonna si riporta il parametro analitico, cui corrispondono i numeri dinorma/metodo riportati nelle colonne successive.I riferimenti normativi sono sempre divisi in quattro classi: EN - ISO - IEC; Metodi Italia-ni (UNI - UNI EN - UNI EN ISO - CEI – NOM); IP; ASTM.Tutti i metodi che durante l’anno hanno avuto revisioni o modifiche sono evidenziati conlo sfondo grigio.La nuova versione dei metodi ASTM è stata confrontata con l’edizione precedente e nelfoglio “Commento alle nuove revisioni” si riportano i risultati di tale confronto. Quandocompare la dizione “equivalente” significa che c’è una perfetta rispondenza tra le metodi-che; differenze non sostanziali tra i vari metodi sono riassunte nell’espressione “tecnica-mente equivalenti”; per i metodi in cui è stata riscontrata anche una sola, ma significativadifferenza, viene riportata l’espressione “non equivalente”. Per i metodi IP si rimanda al sito http://ein.powerweb.co.uk/cssiptmqbe.htm dove èdisponibile l’elenco aggiornato dei metodi e un loro confronto con i metodi ASTM e ISO.Preso atto della velocità di cambiamento dei metodi in ambito normativo, soprattutto deimetodi ASTM, si ricorda che la presente guida, non potendo essere aggiornata in temporeale, ma facendo riferimento ad una valutazione temporale pari a un anno solare, ha dellelacune, insite proprio nella modalità in cui è stato concepito il lavoro di revisione.

M. Sala1*F. Taormina1

P. Fornasari2

P. Ruggeri3

1 INNOVHUB - SSIAzienda Speciale della Camera di Commercio

Divisione SSOG - Milano

2CESI SpA - Engineering &Environment - ISMES Division

Piacenza

3ENI SpA - Refining & MarketingMilano

nota tecnicaLubrificanti

Corrispondenze tra metodi analitici*

(gennaio-dicembre 2012)


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PARAMETRO ANALITICO EN-ISO-IEC IP ASTM DMetodi Italiani


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NotiziarioIN BIBLIOTECA

RIVISTE IN ABBONAMENTO

OLI E GRASSI

dal 1938 European Journal of LipidScience and Technologyeditore: Wiley-Vch Verlag Gmbhperiodicità: mensileThe journal is the official organ of the EuropeanFederation for the Science and Technology ofLipids (Euro Fed Lipid).Lipids, fats and oils play an ever increasing role inmany aspects of health, science and technology,e.g. health requirements, metabolism, tailor-maderaw materials and renewable resources.The European Journal of Lipid Science and Tech-nology focuses on the scientific and geographicalintegration of this varied spectrum ranging fromlipidomics, nutrition and health to analytics,biotechnology and process engineering as well aschemistry and physical chemistry.

dal 1973 Journal of Agricultural andFood Chemistry editore: American Chemical Societyperiodicità: 51 volte l'annoThe Journal of Agricultural and Food Chemistrypublishes studies and researches dealing with thechemistry and biochemistry of agriculture andfood. The Journal also encourages papers withchemistry and/or biochemistry as a major compo-nent combined with biological/sensory/nutritional/toxicological evaluation related to agricultureand/or food.

dal 1954 Journal of the American OilChemists' Society - JAOCSeditore: American Chemical Societyperiodicità: mensileJAOCS is a monthly, peer-reviewed journal devot-ed to fundamental and practical research, produc-tion, processing, packaging and distribution in thegrowing field of fats, oils, proteins and other relat-ed substances.

dal 1992 Oils and Fats Internationaleditore: Quartz Business Media Quartz Houseperiodicità: 8 volte l'annoOils and Fats International (OFI) - internationaljournal devoted to oils and fats.Contents:• Themed biofuel issues• Official catalogue for OFI events• Industry news• Specialist features and articles

• Company news• Legislation, merger and acquisition updates• International market reports• New product information• Diary of events

dal 1990 Lipid Technology editore: Wiley-Vch Verlag Gmbhperiodicità: mensileLipid Technology provides with a monthly source ofconcise information on the critical developmentsand trends in the oilseeds, fats and oils industries;fats and oils markets; as well as lipids in science,technology, biotechnology, food, agriculture, nutri-tion and pharmaceuticals.

COSMETICA

dal 1977 Cosmetics & Toiletrieseditore: GCI Magazineperiodicità: mensileCosmetics & Toiletries magazine provides expertinformation on new technologies, ingredients, inter-national regulations and industry news and analysis.

dal 1999 Kosmeticaeditore: Cosmeo S.R.L.periodicità: 9 volte l'annoLa Rivista fornisce informazioni approfondite sulledifferenti tematiche inerenti le materie prime e gliingredienti, la filiera produttiva, la ricerca e svilup-po, il controllo qualità, il marketing e il manage-ment aziendale. Una specifica sezione è riservata al packaging delsettore e ampio spazio è dedicato all’attualità e almercato italiano.

DETERGENTI E TENSIOATTIVI

dal 2007 Journal of Surfactants andDetergents (JSD)editore: American Oil Chemists Societyperiodicità: bimestraleJSD is concerned with the practical and theoreticalaspects of oleochemical and petrochemical sur-factants, soaps and detergents. This scientific jour-nal publishes peer-reviewed research papers, andreviews related to surfactants and detergents tech-nologies.

dal 1900 Sofw Journal editore: Verlag Fuer Chemische Industrie H. Ziol-kowsky Gmbhperiodicità: mensile(Ex Seifen, Oele, Fette, Wachse)Since over 135 years now the professional journal


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European Biodiesel12-13 June 2013 - Lisbon, PortugalEuropean Biodiesel 2013 will be held in Lisbon, Portu-gal on June 12-13, 2013.European Biodiesel 2013 is a conference that coverstopics such as:• National implementation of EU biofuels policy• The future of biodiesel from the viewpoint of the fos-

sil fuel industry• Policy implementation & uncertainty• Regional industry focus: Eastern Europe• Running a competitive biodiesel plant in Europe• Commercial-scale advanced generation renewable

diesel production• Ensuring a sustainable supply chain• Aviation insight: Jatropha-based bio jet fuel• Emerging feedstock outlook: Cost & availabilityhttp://www.wplgroup.com/aci/conference/eu-eaf6.asp

2013 Conference - Sustainable Agriculture through ICT innovation23-27 June 2013 - Turin Conference Centre Torino, ItalyArgomenti trattati:- ICT applications for food chain and logistics- ICT applications for natural resources management - Environmental information systems/Environmen-

tal management systems- Remote Sensing, GIS applications and planning tools- Robotics in Agriculture- On line farm services and web applications- Social Networking- Cloud computing applications and apps- ICT Applications for sustainable precision farming- ICT applications for sustainable biomass produc-

tion and use- ICT applications for agriculture and sustainability- Weather prediction models for sustainable agri-

cultural production- Decision Support Systems for agriculture- e-agribusiness- Rural economies and ICT policies for rural devel-

opment- Metadata and data standards in agriculture- Extension and knowledge repository services- Agricultural Information Systems including data

banks and data miningPer informazioni:www.efita2013.org

COSI 2013 - 9th Coatings SciencesInternational 201324-28 June 2013 - Noordwijk, The Netherlands Conference Secretariat:Mrs Imanda Scholten, Conference SecretariatCOSI 2013

SOFW Journal tracks the development of the soap,detergent, perfume, personal care and cosmeticindustry in chemistry and technology as well as theprogression of associations and societies.

dal 1964 Tenside Surfactants Detergentseditore: Hanser Carl Verlag Gmbh & Co. Kg-Arvatoperiodicità: bimestraleTenside Surfactants Detergents deals comprehen-sively with all aspects of the surfactant chemistrysuch as: synthesis, analysis, physiochemical prop-erties, new classes of surfactants, progress in pro-duction processes, application-related problems,environmental behavior.

VERNICI

dal 2008 EC European Coatings Journal editore: Vincentz Network GMBH & CO.KGperiodicità: mensileEuropean Coatings Journal serves Europe as asource of information for the coatings, printing inks,construction chemicals, adhesives and sealantsindustry.

dal 2004 Progress in Organic Coatingseditore: Elsevier Science S.A.periodicità: mensileThe aim of this international journal is to analyzeand publicize the progress and current state ofknowledge in the field of organic coatings andrelated materials. The journal publishes papersdealing with such subjects as:• Chemical, physical and technological properties

of organic coatings and related materials• Problems and methods of preparation, manufac-

ture and application of these materials• Performance, testing and analysis.

CONGRESSINo

tiziar

io


- Lipidomics - Jonathan Markham, Donald DanforthPlant Science Center, St. Louis, USA. "Solving theRiddle of Sphingolipids through Lipidomics.

http://www.eurofedlipid.org/meetings/bordeaux2013/index.php

Institute of Food Technologists’Annual Meeting and Expo13-17 July 2013 - Chicago, Illinois, USA Gli argomenti che verranno discussi durante ilMeeting Annuale IFT verteranno sugli ultimi svilup-pi scientifici, le più recenti innovazioni e le ultimetendenze. Professionisti del settore alimentare siincontreranno per uno scambio di informazioni. Per informazioni:www.ift.orghttp://www.am-fe.ift.org/cms/

PREP 2013, 26th Internatational SymposiumPreparative & Process Chromatography14-17 July 2013 - The Westin Boston Waterfront,Boston, MA, USAI maggiori esperti scientifici, produttori e media alivello internazionale, si incontreranno e presente-ranno le idee più innovative nelle aree della croma-tografia preparativa e di processo mostrandocome gli ultimi prodotti e tecnologie guideranno infuturo l’innovazione nella ricerca scientifica.Gli obiettivi di questi incontri saranno quelli di:- Trovare delle soluzioni di separazione cromato-

grafica- Ottimizzare i processi della cromatografia per

una migliore produzione in termini di economia- Discutere dei cambiamenti attuali e futuri e le

opportunità per i produttori così come per imedia e i fornitori di attrezzature

- Dare una visione generale degli ultimi prodotti etecnologie che sono alla guida dell’innovazionenella cromatografia preparativa

- Interagire a livello internazionale con espertiscientifici ed ingegneri per tutti gli aspetti dellacromatografia liquida preparativa e di processo

Per aggiornamenti:http://www.prepsymposium.org/

36th Annual Short Course: Advances inEmulsion Polymerization5-9 August 2013 – Davos, Switzerland www.davoscourse.com

XV Latin American Congress andExhibition on Fats and Oils20-23 August 2013 - Sheraton Santiago Hotel and Con-vention Center Santiago, Chile Il Convegno sarà caratterizzato da:- importanti temi riguardanti l’industria degli oli e

dei grassi

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NotiziarioEindhoven University of TechnologyHelix STO 2.34, P.O. Box 513,5600 MD Eindhoven The NetherlandsTel: + 31 40 247 2770, Fax:+31 40 244 5619www.coatings-science.comEmail: [email protected]

XXXV CIOSTA & CIGR V Conference 3-5 July 2013 - Denmark Le conferenze CIOSTA si concentreranno sulla bio-gestione della produzione e l’organizzazione dellavoro, sulla base di approcci di ingegneria e tec-nologie innovative.Argomenti:- Service robots in bio-production system- Field machinery management- Traceability system- Biomass and agri-food supply chains- Decision support system- System analysis and design- Precision farming based operations planning- Processing, post-harvest technology- Liverstock management- Labour, ergonomist,safety and healthwww.ciosta.org

6th European Symposium on PlantLipids7-10 July 2013, Bordeaux, Franceil 6 ° Simposio europeo dei lipidi vegetali organizza-to da Euro Fed Lipid si terrà a Bordeaux, Francia, dal7 al 10 luglio, 2013. Il programma scientifico saràcaratterizzato da sessioni orali che riguarderanno lediverse aree di ricerca dei lipidi. Il simposio forniràopportunità di scambio e collaborazioni.Programma scientifico (aggiornato al 12.11.2012] Argomenti principali: - Fatty Acid Modification - Frederic Beaudoin,

Rothamsted Research, Harpenden, Herts, UK - Membrane and Chloroplast Lipid Metabolism -

Peter Dörmann, University of Bonn, Germany - Storage Lipid Accumulation and Mobilisation -

Sebastian Baud, Institut Jean-Pierre Bourgin,Versaille, France. "Importance of TranscriptionalRegulators for the developmental Control of FattyAcid Production in Arabidopsis thaliana

- Plant Lipid Biotechnology - Johnathan Napier,Rothamsted Research, Harpenden, Herts, UK

- Lipid Surface and Suberin Metabolism - Ljerka Kunst,University of British Columbia, Vancouver, Canada

- Lipid Signalling - Ingo Heilmann, University ofHalle-Wittenberg, Germany

- Sphingolipids and Sterols - Howard Riezman,University of Geneva, Switzerland. "Sterol andSphingolipid Homeostasis and Function"

- Intracellular Lipid Trafficking - V. Zarski. "Mem-brane Lipid Dynamics in the Regulation of PollenTube Expansions"


128


Notiz

iario - Dissecting lipid metabolism in diabetes and ath-

erosclerosis- Lipid metabolism, transcription, neurogenesis,

stemness- Phospholipid metabolism in physiology and dis-

ease- Lipids and membranes in stress management:

perception, signaling and adaptation.Per aggiornamenti aggiornamenti: http://www.icbl.unibe.ch/index.php?id=81

10th Oilseed & Oil Processing ShortCourse17-18 September 2013 - Munich GermanyI corsi brevi offriranno ampi programmi che copro-no una vasta gamma di argomenti con specificiaspetti pratici:- Aquaculture feed;- Snackfoods;- Soyfoods;- Functional Lipids; - Functional Beverages;- Advanced Oil Processing;- Industrial Oil UsesDestinatari dei corsi:tecnici di prodotto, ricerca e sviluppo, autorità divigilanza, ingegneri, tecnici, ingegneri di progetto,manager dei processi di sviluppo del business, divendita e specialisti di marketing, produttori diapparecchiature, della formulazione del prodotto,costruttori di impianti, trasformatori, chimici. Per informazioni:http://www.smartshortcourses.com/

PT CONF - 4th International Proficiency Testing Conference18-20 September 2013 - Brasov, RomaniaTemi della Conferenza:- Reference materials and Proficiency Testing (RM)- Validation of Testing Methods and Proficiency

Testing (VTM)- Uncertainty of Measurement and Proficiency

Testing (UM)- International Quality Control of Analyses and Pro-

ficiency Testing (IQC)- Metrology, Traceability and Proficiency Testing

(MT)- Accreditation of Quality Management and partic-

ipation at Proficiency Testing (AQM)- Development of the Analyses Methods in the

Testing Laboratories (DAM)Organizzato da:Ceprocim S.A.Research, Consulting & ProcessDevelopment,www.ceprocim.roAFSIC Association of Proficiency Testing Providershttp://www.afsic.ro/about_en.htmlIn co-operation with:

- significativa opportunità ai leader dell'industria eal mondo accademico per discutere le opportu-nità esistenti oggi sul mercato

- sessioni di poster e presentazioni orali di espertileader nel settore

Annotare l’evento. Per informazioni e aggiorna-menti:AOCS Meetings Department Phone: +1 217-359-2344Fax: +1 217-351-8091 http://lacongress.aocs.org/en/index.cfm

10th AIIA Conference – Horizons inagricultural, forestry and biosystemsengineering8-12 September 2013 – Viterbo (Italy)Argomenti:- Forest-wood chain- Hydrology and dynamics of water and sediments

in agricultural and mountain basins: monitoring,modeling and risk analysis

- Hydraulics and hydro-morphological processesfor stream and river restoration and management

- Information technology, automation and preci-sion farming and forestry

- Structures and technologies for livestock produc-tion: technical, energy and environmentalaspects

- Structures and technologies for protected cropproduction: technical, energy and environmentalaspects

- Post harvest, food and process structures andtechnologies

- Power and machinery in agriculture and forestry- Analysis, modeling and planning of rural areas- Renewable energy, biomass and biological

residues- Safety, health and ergonomics - SHWAnet inter-

national meeting- Sustainable planning and management of soil

and water resourcesSegreteria organizzativa:[email protected]

246th American Chemical SocietyNational Meeting & Exposition8-12 September 2013 - Indianapolis, Indiana, USA www.acs.org

54rd International Conference on theBioscience of Lipids17-21 September 2013 - Bari, Italy Argomenti del Simposio:- Signal transduction, gene expression and circa-

dian rhythm in the regulation of lipid metabolism- Nuclear receptors and the transcriptionalregulation of lipid metabolism

- The gut-liver axis route for lipids


ri di impianti di riempimento. Il successo della formula è evidente anche dalnumero crescente di visitatori internazionali. L’edizione 2011 è stata l’inizio di numerosi progettiinternazionali alla ricerca di processi più efficientiper mantenere il rapporto prezzo/prestazioni sem-pre su livelli ottimali. Massima resa e minimo sprecodi materie prime, unitamente al risparmio energeticonel processo di lavorazione, sono i fattori fondamen-tali. A questi si aggiungono considerazioni legatealla salute: le scoperte scientifiche nella ricerca ali-mentare impongono una gestione precisa e attentadelle tecniche di raffinazione, per evitare la forma-zione di sostanze nocive di recente scoperta o elimi-narle del tutto. Quando si parla di singole macchine,la tecnologia punta a capacità sempre più elevate,ad esempio con presse in grado di trattare fino a800 tonnellate di semi al giorno. Soluzioni specifi-che, su misura per le esigenze di ciascun cliente,sono quindi fondamentali per il settore.L’industria di lavorazione di oli e grassi sta prestan-do grande attenzione all'impiego di enzimi, finaliz-zato a migliorare determinate reazioni, ad aumen-tare le rese e a ottimizzare l'efficienza dei processi,ad esempio nei trattamenti di demucillaginazione,esterificazione e transesterificazione. I risultati diquesto lavoro dovrebbero essere visibili a oils+fats2013. A questi si aggiungono esperimenti che,sfruttando tecniche di miscelazione intensiva, adesempio con miscelatori a cavitazione, puntano adaumentare le reazioni, e quindi le rese, e a ridurreal minimo gli sprechi nei processi. Come già acca-de in molti altri settori, anche nell’industria degli olialimentari si denota una chiara tendenza verso lariduzione degli sprechi e dei consumi energetici.

La dieta mediterranea favorisce l’industria degli olialimentariL’olio è uno degli alimenti chiave della dieta medi-terranea, sempre più apprezzata nel mondo, chefavorisce una crescita costante e continua del set-tore degli oli alimentari. Le proprietà benefiche,soprattutto dell’olio di oliva, sono indiscutibili, gra-zie alla presenza di acidi grassi polinsaturi, minera-li preziosi e vitamine. L'olio d'oliva di provenienzaspagnola, ad esempio, che rappresenta oltre i dueterzi della produzione mondiale insieme a Italia eGrecia, viene esportato prevalentemente in botti-glie di vetro per esaltarne le caratteristiche di pro-dotto da "gourmet". Queste bottiglie vengono riem-pite con sistemi innovativi come quelli che sipotranno vedere esposti a drinktec 2013. Il livellodi riempimento esatto viene garantito con l’uso dicelle di pesatura posizionate in corrispondenza diogni valvola di riempimento, sfruttando il principiodi gravità, senza alcuna pressione. Per il confezio-namento di oli alimentari in contenitori di PET, inve-ce, sono state sviluppati nuovi sistemi di riempi-

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Notiziario“Transilvania” University from Brasov Faculty ofWood Engineeringhttp://www.unitbv.ro/il/enLucian Blaga University from Sibiu “HermannOberth” Engineering Facultywww.ulbsibiu.ro/en/Conference secretariat: Ceprocim S.A., Bucarest,Romaniae-mail: [email protected]

Home and Personal Care IngredientsMiddle East and Eurasia Exhibition &Conference18-19 September 2013 – Istanbul, TurkeyE’ un evento importante focalizzato su prodotti eservizi per l'industria cosmetica e domestica.La Fiera HPCI offre ai visitatori informazioni sunuovi servizi e prodotti.Piattaforma ideale per confrontarsi con i rappresen-tanti, fornitori di prodotti, tecnici di laboratori, servi-ce provider su problematiche inerenti il settore.Due giorni di Conferenza scientifica per fornire leultime importanti informazioni sugli sviluppi in R&D,nuove formulazioni e problematiche normative inmateria di cosmetici e detergenti.http://www.hpci-congress.com/

OILS+FATS 201318-20 Settembre 2013 – Monaco di BavieraL’industria degli oli e dei grassi si dà appuntamen-to nelle date dal 18 al 20 settembre al Centro Fieri-stico di Monaco di Baviera. Al salone oils+fatssono attesi circa 60 espositori da oltre 10 Paesi eun migliaio di visitatori qualificati da tutta Europa.La fiera specializzata per le tecnologie e il com-mercio di sostanze oleose e grasse, si avvicina allaquinta edizione con una nuova immagine e un'of-ferta ancora più ricca. Espositori da oltre dieciPaesi presenteranno macchine, sistemi, compo-nenti e mezzi ausiliari per la produzione e la lavo-razione di oli e grassi. Alle tecnologie si affiancanomaterie prime e ingredienti, soluzioni logistiche,sistemi di riempimento e confezionamento. Lapeculiarità di quest’anno è lo svolgimento dioils+fats, negli stessi giorni di drinktec, con moltesinergie e opportunità di interazione e contatto conil comparto degli alimenti liquidi.Drinktec, salone internazionale delle tecnologieper le bevande e gli alimenti liquidi, è la fiera piùimportante del settore. A drinktec 2013, sono atte-si circa 1.500 espositori da oltre 70 Paesi e circa60.000 visitatori da più di 170 nazioni.L’abbinamento a drinktec, è una scelta mirata inquanto in questo modo è stata ampliata l'offertaespositiva. I visitatori non trovano più in fiera solosocietà di ingegneria e costruttori di macchineesclusivamente specializzati nel settore degli oli edei grassi, ma anche fornitori di pompe o produtto-


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Notiz

iario mento monoblocco che uniscono la produzione in

linea di PET con sistemi di riempimento a bilancia.I vantaggi dei blocchi, oltre al grado elevato di igie-ne, sono il forte risparmio energetico, la maggioreefficienza e la riduzione dei costi di manutenzione,oltre alla possibilità di utilizzare bottiglie leggere.

Olio di colza e materie prime rinnovabiliI tassi di crescita sostenuti nel comparto degli olialimentari erano in primo piano già a oils+fats2011. Il secondo grande tema era il biodiesel, chenegli ultimi anni ha registrato un ulteriore aumentodi interesse. Il mercato del biodiesel resta lo sboc-co principale per le coltivazioni di colza tedesche.Le aziende dotate di impianti integrati, che partonodall’estrazione e dalla raffinazione dell’olio per arri-vare alla produzione di oli alimentari e biodiesel,sono sopravvissute al processo di consolidamentoe continueranno a crescere. Chi produce esclusi-vamente biodiesel, invece, è costretto a rincorrerela coltivazione e la produzione di materie prime,spingendosi in America Latina, in particolare inArgentina e Brasile con le loro gigantesche pianta-gioni di soia, piuttosto che nel Sudest asiatico dovedominano le palme da olio o nelle regioni del Cana-da dove si coltiva la canola.La grande varietà di impiego dell’olio di Raps, al dilà della produzione di biodiesel, consente moltepli-ci utilizzi innovativi ed ecologici. Ad esempio, èstato sviluppato un asfalto nel quale il petrolio èstato sostituito per il 20 percento con olio di colza.Un produttore di membrane per tetti e impermeabi-lizzazioni ha sostituito il bitume con olio di colza,che ha trovato applicazione anche come separato-re per il cemento nell’industria edilizia. Il metileste-re di colza viene utilizzato anche nell'industria dellevernici, nella fabbricazione di automobili e nellaproduzione di carta autocopiante. È questo che siintende per sostenibilità.In futuro potrebbe acquisire rilevanza la lavorazio-ne delle alghe per la produzione di biocarburanti,proteine, oli e grassi. Le materie prime rinnovabilisono sotto molti punti di vista un tema di grandeattualità per l’alimentazione e l’energia. Il saloneoils+fats 2013 è il posto giusto per aggiornarsisulle ultimissime novità.Per maggiori informazioni: http://www.oils-and-fats.com/en

drinktec 201316-20 settembre 2013 - Monaco di Baviera drinktec, salone internazionale delle tecnologieper le bevande e gli alimenti liquidi, una fieraimportante del settore. Produttori (fornitori) di tuttoil mondo, dalle multinazionali alle medie imprese,si incontrano a Monaco di Baviera con i grandiproduttori e distributori di bevande e liquid food.

drinktec viene considerato dagli addetti ai lavorila principale vetrina mondiale per la presentazio-ne di novità. Le aziende produttrici propongono letecnologie più recenti per la produzione, il riempi-mento e il confezionamento di tutti i tipi di bevan-de e alimenti liquidi, accanto a materie prime esoluzioni logistiche. L’offerta del salone è comple-tata dai temi legati al marketing delle bevande eal design delle confezioni. Circa 1500 espositori da 70 Paesi presenterannol'intera gamma di tecnologie per bevande e liquidfood, incluse le soluzioni per imballaggio e confe-zionamento e gli strumenti di marketing. Il 60 per-cento circa delle aziende espositrici proviene dal-l'estero: drinktec vanta un livello di internazionali-tà superiore a qualsiasi altra manifestazione, intermini sia di espositori sia di visitatori. La doman-da di superficie espositiva è altrettanto consisten-te e, a oggi, già il 90 percento dell’area disponibi-le è stata assegnata. A settembre sono attesi oltre60.000 operatori da tutto il mondo.UNA SOLUZIONE PER OGNI PROBLEMA E OGNI ATTIVITA’“Produzione + Riempimento + Confezionamento+ Marketing” è il nuovo claim che parla chiaro:drinktec è molto più di una vetrina tecnologica. Lamanifestazione abbraccia tutto ciò che riguardal'industria delle bevande e degli alimenti liquidi alivello mondiale, come sottolinea la direttrice delsalone Petra Westphal: “Si parla di materie primecome di processo di riempimento, ma anche diimballaggio, confezionamento e marketing, tuttiargomenti che occupano un posto di primopiano.” Oltre a essere una vetrina di tecnologie e novità,drinktec si propone come una vera e propria“borsa di idee e informazione”, un punto di incon-tro privilegiato per la comunicazione e le relazio-ni. Per questo motivo a Monaco di Baviera sidanno appuntamento operatori specializzati ditutti i comparti del settore. In fiera sono presentitutte le tipologie di aziende, dai produttori dibevande e alimenti e società commerciali interna-zionali ai piccoli e medi birrifici che operano suscala regionale.Gli espositori distribuiti in dodici padiglioni hannosoluzioni adatte per qualsiasi problema e perimprese di tutte le dimensioni, dalla linea di riem-pimento completa al singolo prodotto. Su un’areadi 132.000 metri quadrati verranno presentatiimpianti, sistemi e prodotti per la produzione, ilriempimento, il confezionamento e il marketing,incluse le materie prime e le soluzioni di logistica.- L’area più estesa è quella dedicata alla tecnica

di processo specifica per la produzione/lavora-zione di bevande, latte e alimenti liquidi. Questocomparto da solo copre una superficie di circa80.000 metri quadrati, che comprende anchePETpoint, un intero padiglione dedicato alle tec-


zione, il riempimento e il confezionamento diacque, succhi e bibite.- Imperdibile per gli addetti al marketing e allo

sviluppo dei prodotti di grandi e piccole azien-de la “Special Area New Beverage Concepts“nel padiglione B1, dove verranno presentatenuove strategie per dolcificanti, coloranti earomi.

- Come si evolve il mercato, non solo dal punto divista tecnologico, ma anche in termini di marke-ting? A questa domanda risponderà il drinktec-Forum nel padiglione A2, con un programma dipresentazioni dedicate ad esempio al tema del-l’Audio Branding. Inoltre, nell’Innovation FlowLounge la tecnologia incontrerà il marketing.

- Il prestigioso Beverage Forum organizzato dallarivista specializzata Beverage World avrà luogoalla vigilia di drinktec (la domenica che precedel’apertura del salone), per la prima volta fuoridagli Stati Uniti, configurandosi quasi come unasorta di mostra speciale. Nell’ambito del forumsi incontreranno i CEO delle grandi multinazio-nali per discutere del presente e del futuro del-l’industria delle bevande con i massimi rappre-sentanti delle aziende fornitrici. Un appunta-mento assolutamente imperdibile.

- In occasione di drinktec 2013, le bevande piùinnovative verranno premiate dalla casa editricebritannica Foodbev Media con i Beverage Inno-vations Awards.

• Focus sulla birra Con o senza alcool, birra vera e propria o bevan-da analcolica: i confini si fanno sempre più sfuo-cati, le bevande miste sono in voga in tutto ilmondo e, sempre più spesso, i produttori di bibi-te sono anche produttori di birra, e viceversa.Inoltre, la birra registra tassi di crescita elevatissi-ma in mercati emergenti in Asia e Africa. Nei mer-cati occidentali maturi, invece, furoreggiano i bir-rifici che propongono ricette speciali. Per questomotivo i birrifici rappresentano per gli espositori didrinktec un target importante tanto quanto i pro-duttori di bevande analcoliche. Circa due terzidelle aziende propongono infatti soluzioni per bir-rifici di tutte le dimensioni: drinktec 2013 offriràquindi una risposta a tutte le esigenze anche inquesto settore. Il salone metterà in mostra tutta lagamma di tecnologie di processo per birra emalto.- Nella seconda giornata del salone verranno

offerte in degustazione le birre premiate con la“European Beer Star”, uno dei riconoscimentipiù importanti per il mondo della birra a livellomondiale. Fra tutte le birre verrà eletta la “prefe-rita del pubblico”.

- Il campionato mondiale dei sommelier dellabirra metterà alla prova le capacità dei più gran-di esperti mondiali. La gara si svolgerà la dome-

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Notiziarionologie per il PET. Un settore sempre più impor-tante e ampio all’interno della tecnica di proces-so è rappresentato dai sistemi di riempimento econfezionamento.

- Il settore trasversale della tecnica di processo,dell’automazione, della tecnica di controllo ecomando e dell’IT è raccolto nei Padiglioni A3,A4 e B2, su una superficie complessiva di circa25.000 metri quadrati.

- Il padiglione B1 e una porzione del B2 sonoriservati a materie prime, additivi e sostanze dilavorazione, che assumono sempre maggioreimportanza. Si troveranno, ad esempio, sistemidolcificanti, coloranti e aromi, e ingredienti pernuovi tipi di bevande di ogni genere. Quest’areaespositiva non è interessante solo per gli esper-ti di tecnologie: anche gli addetti al marketing eallo sviluppo di nuovi prodotti troveranno spuntiinteressanti e avranno la possibilità di provarebevande particolari e innovative.

- Altrettanto interessanti per chi opera nel marke-ting e nella comunicazione i padiglioni A1 e A2.Accanto a soluzioni innovative per il confeziona-mento e l'etichettatura, ci sarà tutto quantoserve al mondo della gastronomia, in particola-re per la promozione e la pubblicità.

• Temi chiave: sostenibilitàAnche nell’industria delle bevande, come in altrisettori, tutto ruota attorno al grande tema dellasostenibilità. La tecnica di processo innovativa invetrina a drinktec 2013 punta in primo luogo allariduzione dei consumi di energia e acqua, con ilfine ultimo di ottimizzare la redditività e l'economi-cità della produzione. Anche sul fronte degliimballaggi l'obiettivo è massimizzare l'efficienzadi impiego delle risorse, principalmente attraver-so la riduzione degli spessori dei materiali delleconfezioni. Altri temi generali che attraverserannotutti gli stand a drinktec 2013 sono la flessibilitàdegli impianti, la progettazione igienica e l’assicu-razione della qualità.• Focus su bibite e bevande analcolicheNegli ultimi anni pochi comparti dell’industriadelle bevande hanno conosciuto uno sviluppodinamico come le bibite analcoliche. Questacategoria di bevande è particolarmente soggettaalle tendenze di consumo. Il numero di novità enuovi prodotti che il mercato propone ogni anno èenorme. Per chi fornisce tecnologie, questo signi-fica che bisogna essere molto reattivi e pronti adadeguarsi alle evoluzioni del mercato. La flessibi-lità è fondamentale soprattutto nell’ambito delconfezionamento. Anche a drinktec le tecnologie, le soluzioni diimballaggio e gli ingredienti per le bevande anal-coliche sono in primo piano da molti anni. Il pros-simo settembre, circa due terzi degli espositoriproporranno tecnologie specifiche per la produ-


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Notiz

iario Nei padiglioni si trova infatti tutto ciò che serve

per trasformare un prodotto in un marchio: ingre-dienti, soluzioni di imballaggio, idee per l’etichet-tatura e, non ultimo, strumenti di promozione epubblicità. Dove se non a drinktec i tecnici posso-no incontrarsi e scambiare idee e punti di vistacon gli addetti al marketing? - Girando per i padiglioni di drinktec 2013, le

menti creative degli uffici marketing dei produt-tori di bevande e alimenti liquidi potranno rac-cogliere spunti e suggerimenti utili. In particola-re, nell’area “World of Labels” potranno aggior-narsi su design e tendenze di imballaggio, eti-chettatura e pubblicità di ogni genere.

- La “Special Area New Beverage Concepts”(pad. B1) mette in vetrina ingredienti per lacreazione di bevande innovative.

- Un’altra novità di drinktec è la Die Innovation-FlowLounge, una sorta di "borsa del networkinge dei contatti" per gli addetti al marketing cheguardano oltre la loro sfera di competenza perinteragire con rappresentanti di altri reparti del-l'impresa. Scambio di idee, raccolta di spunti esuggerimenti, valutazione di nuove proposte:questo e altro nella lounge all'ingresso ovest(Eingang West). Ogni giorno verranno propostitemi sempre diversi, con brevi presentazioni etavole rotonde. E quando serve una pausa direlax dai ritmi frenetici della fiera, basta acco-modarsi a uno dei tanti bar dove si possonogustare bevande innovative.

- Come si evolve il mercato, non solo dal punto divista tecnologico, ma anche in termini di marke-ting? A questa domanda risponderà il drinktec-Forum nel padiglione A2, con un programma dipresentazioni dedicate ad esempio al tema del-l’Audio Branding.

Per maggiori informazioni:www.drinktec.comhttp://www.drinktec.com/en/Home

15th AOCS Latin American Congressand Exposition on Fats and Oils20-23 September 2013 - Santiago, ChileLe aree tematiche generali saranno le seguenti:• Analytical techniques and applications• Biotechnology of lipids• Chemistry and lipid synthesis• Detergent and soaps• Lipid oxidation and antioxidants• Norms, regulations and food safety• Oils, fats and lipids in human and animal health

and nutrition• Production statistics of fats and oils• Specialty fats and oils• Technological development and innovations in

the processing of oil seeds, fats and oilshttp://lacongress.aocs.org/en/index.cfm

nica antecedente all'apertura della manifesta-zione. Giunto alla terza edizione, il campionatomondiale dei sommelier della birra è organizza-to dall’accademia Doemens.

- La prima giornata del drinktec-Forum sarà nelsegno della birra, con un programma di inter-venti dedicati ad esempio al confezionamento,particolarmente interessante per i visitatori chesi occupano di marketing.

• Focus sul liquid foodLe tecnologie per gli alimenti liquidi, in particola-re per il latte e le bevande a base di latte, hannoacquisito negli ultimi anni sempre maggior impor-tanza nell’ambito di drinktec. Il motivo è cheanche in questo comparto si assiste a una cre-scente contaminazione, con la creazione dibevande miste a base di latte e succhi di fruttae/o bibite analcoliche. Gli espositori di drinktechanno risposto anche a questa evoluzione delmercato. Quasi un espositore su due presente infiera fornisce tecnica di processo specifica per lalavorazione del latte. Rispetto alla scorsa edizio-ne, il comparto è aumentato del dieci percentocirca.- La ricca offerta di soluzioni di confezionamento

e logistica a drinktec richiama operatori anchedal mondo del non-beverage e degli alimentiliquidi.

- Le principali associazioni hanno riconosciuto lacrescente importanza di drinktec per l’industriadel latte. L’associazione centrale dell’industriadel latte tedesca (Zentralverband DeutscherMilchwirtschaftler-ZDM) e l’associazione euro-pea AEDIL “Association Européenne des Diplô-més de l`Industrie Laitière” sfruttano drinktec2013 per il networking e lo scambio di idee eopinioni. Inoltre, i membri delle associazioni ter-ranno le rispettive assemblee durante drinktec.

- Nella giornata di venerdì si raccomanda ai rap-presentanti dell’industria del latte di fare unsalto al drinktec-Forum, dove sono in program-ma numerosi interventi dedicati al latte e agli ali-menti liquidi. Ricordiamo che tutti i convegni didrinktec-Forum sono compresi nel biglietto diingresso al salone e sono coperti da traduzionesimultanea in inglese e tedesco.

LA TECNOLOGIA INCONTRA IL MARKETINGIl successo di un prodotto dipende anche dall’ori-ginalità del messaggio commerciale: in due paro-le, da un marketing azzeccato. I brand managersono figure decisionali sempre più importanti,perché in ultima analisi un prodotto deve esserevenduto. Gusto, colore, confezione, etichetta...affinché tutto questo si traduca in un successocommerciale, servono tecnologie, ingredienti eadditivi adeguati. drinktec è quindi un appunta-mento imperdibile non solo per i tecnici, maanche per gli esperti di marketing delle aziende.


American Association for Aerosol Research (AAAR) 32nd Annual Conference30 September-4 October 2013 - Oregon ConventionCenter Portland, Oregon, USA Il meeting offre una significativa opportunità perpromuovere e divulgare i progressi tecnici nelcampo della ricerca aerosol, fornendo un'eccellen-te piattaforma internazionale di scambio tra esper-ti nel campo della ricerca e dell'industria.Al momento non sono disponibili informazioni sulprogramma scientifico e sulle modalità d’iscrizione. http://www.aaar.org/

60th Sepawa Congress and EuropeanDetergents Conference08-11 October 2013 - Fulda, Germanyhttp://www.sepawa.com/

3rd MS-Food Day9-11 ottobre 2013, Trento, ItaliaDopo il successo della prima e seconda edizione èun piacere annunciare la 3a edizione MS-Food Day,che si terrà nei giorni dal 9 all’11 ottobre 2013,nella città di Trento, organizzata dalla FondazioneEdmund Mach e dalla Divisione di Spettrometria diMassa della Società Chimica Italiana.Oggi, diventano sempre più importanti alimentisicuri e di alta qualità, con buone caratteristichenutrizionali e sensoriali.La caratterizzazione di componenti alimentari, l'au-tenticazione e la tracciabilità dei prodotti alimenta-ri, il controllo di qualità, l'identificazione e la quan-tificazione di additivi, allergeni, contaminanti chimi-ci e microbiologici, la conservazione di componen-ti alimentari durante lo stoccaggio e la trasforma-zione, le tecnologie per il packaging, le determina-zioni nutrizionali e le proprietà sensoriali, hannoconseguenze diffuse in materia di agricoltura,industria e sanità.La Spettrometria di massa gioca un ruolo fonda-mentale in tutti questi aspetti. In effetti, i migliora-menti tecnologici e metodologici della spettrome-tria di massa e il suo accoppiamento con processidi separazione molecolare resa altamente sensibi-le, metodi veloci e validati sono strumenti fonda-mentali nel campo della scienza e della tecnologiaalimentare.La 3a Conferenza MS-Food Day è un'ottima occa-sione per presentare lo stato di avanguardia dellaspettrometria di massa in chimica degli alimenti,insieme con le più recenti innovazioni nella stru-mentazione e nelle applicazioni. Occasione ottima-le per soddisfare le esigenze e le offerte delle isti-tuzioni accademiche, industrie alimentari e di stru-mentazione.Gli argomenti principali sono:- Innovations in food science applications of MS

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NotiziarioSciX 2013The Great SCIentific eXchangeSeptember 29 – October 4, 2013 Milwaukee, Wisconsin, USA Sostenibilità e minimizzazione dell'impatto ambien-tale sono diventati i driver di innovazione ampia-mente riconosciuti nel settore della chimica.Aree tematiche:- Atomic Spectroscopy - Surface Science- Bioanalytical Analysis- Chemometrics- Nanotechnology- Process Analytical Chemistry- Vibrational Spectroscopy- Pharmaceutical Analysis- Chromatography- LIBS- Mass Spectrometry- Electrophoresis & Microfluidics- Raman Spectroscopy- Low-Field NMR- Security & Forensics- Surface Plasmon Resonance- Microscopy- Biomedical & Clinical Analyses- Environmental Analysis- Separation Science- Agricultural and Food Chemistry- Chemical ImagingSono comunque accettate presentazioni riguar-danti tutti i settori della chimica analitica.Il programma tecnico sarà caratterizzato da ses-sioni orali e sessioni poster.http://www.scixconference.org/scix-home/at-a-glance

Practical Short Course on VegetableOil Processing and Products of Vege-table Oil / BiodieselSeptember 29-October 4 2013 – Texas A&M, CollegeStation, Texas, USAQuesto breve corso è un must per chiunque siacoinvolto in materia di trattamento di olio vegetalee interessato agli ultimi sviluppi in decolorazione,idrogenazione, interesterificazione, deodorizzazio-ne, produzione di biodiesel e grassi trans free.L’obiettivo del corso è formare il personale di pro-duzione in principi e pratiche di: - Nuovi metodi di raffinazione di olio vegetale e di

trasformazione- Metodi di decolorazione, idrogenazione, intere-

sterificazione e deodorizzazione degli oli vegeta-li più importanti

- Produzione di biocarburanti- Produzione di grassi trans free- Sistemi di filtrazione.Per informazioni aggiornate:http://foodprotein.tamu.edu/fatsoils/scvegoil.php


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Notiz

iario gli enti governativi stanno sviluppando l'uso dei

biocarburanti derivati ??dalle alghe per ridurre leemissioni di carbonio dalle centrali elettriche egenerare carburanti rinnovabili nei trasporti.Il Global Algae Biodiesel World 2013 esamina ilvasto potenziale mercato mondiale di biocarburan-te prodotto dalle alghe. Esplora la tecnologia,nuove ricerche, e la conoscenza per lo sviluppo diquesta nuova generazione di biocarburanti. Offreuna singola piattaforma, le migliori competenzeper discutere e analizzare le dinamiche presenti efuture dal punto di vista tecnologico ed economico.Programma del workshop:Il passato, presente e futuro della produzione dellealghe.• Commercializing Algae Biodiesel: Prospects and

Priorities • Algae Growing, Harvesting and Extraction Tech-

nologies • Scaling Up Algae Production to Commercially

Viable Levels • Optimizing Efficiency in Algae Harvesting and

Dewatering • Identifying and Creating the Ideal Strain • Algae Carbon Values: Perspectives from The

Carbon Market • Developing a low cost novel & High Productivity

Enclosed Hybrid System for algae farming for oil • Demonstration of Algae Photo bioreactor and

Biodiesel MakingPer iscrizioni, informazioni e aggiornamenti con-tattare: [email protected]

11th Euro Fed Lipid Congress27-30 October 2013 - Antalya, Turkey Il congresso affronterà temi diversi riguardanti igrassi, gli oli di colza, palma, girasole.I partecipanti provenienti dall'industria, dalla ricer-ca e da associazioni di consumatori, si incontre-ranno per discutere le conoscenze e le applicazio-ni in ampia gamma di discipline correlate ai lipidi.Programma scientifico (aggiornato ad agosto2012)Conferenze plenarie• European Lipid Science Award Lecture • European Lipid Technology Award Lecture • Chevreul Medal Lecture • Wilhelm Normann Medal Lecture • H.P. Kaufmann Memorial LectureArgomenti principali• Analytics, Authenticity, Lipidomics • Health and Diseases • Bioscience, Biocatalysis, Biochemistry • Physical Chemistry • Processing • Oxidation, Antioxidants and Deep Frying • Oilseeds, Plant Breeding and Plant Lipids • Lipids in Animal Science

- Food safety- Quality control/Traceability- Food integrity- Functional food- Metabolomics/Foodomic- Sensory quality: linking sensory and MS data- Packaging- Process monitoring- Methodological and instrumental developments- Recent trends in mass spectrometry- Ion sources and mass analysers- High resolution mass spectrometry- Ambient Mass Spectrometry- Direct Injection/Infusion Mass-pectrometry- High-throughput techniques- Data AnalysisLa Conferenza sarà caratterizzata da sessioni oralie sessioni di poster.Per informazioni aggiornate:http://eventi.fmach.it/MS-Food-Day

American Fats & Oils AssociationAnnual Meeting9-10 October 2013 - Grand Hyatt, NYC, NY Annotare l’evento. Per informazioni e aggiorna-menti:http://americanfatsandoilsassociation.com/

Home and Personal Care IngredientsCentral and Eastern Europe Exhibition & Conference15-16 October Warsaw, PolandDue giorni di Convegno scientifico e seminari tec-nici per fornire le informazioni su nuovi servizi eprodotti; piattaforma ideale per confrontarsi con irappresentanti, fornitori di prodotti, tecnici di labo-ratori, service provider su problematiche inerentiil settore.Per informazioni: http://www.hpci-congress.com/

Global Algae Biodiesel World 20132 Day Algae Fuel State of Art Interna-tional Workshop25-26 October 2013 - Jaipur, IndiaLe Alghe sono una delle più promettenti fonti dibiodiesel, combustibile biodegradabile che puòessere prodotto utilizzando mare e acque refluesenza sfruttare le risorse di acqua dolce.Gli argomenti sono accuratamente selezionati edesaminano gli aspetti biologici, di ingegneria, dimarketing e finanziari per la commercializzazionedelle alghe.Si tratta di una importante piattaforma per gliscambi produttivi tra le comunità accademiche,commerciali e di investimento.Guidati da questioni legate al cambiamento clima-tico e l'aumento del costo del petrolio, le aziende e


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novembre 2013.Una conferenza di due giorni dove esperti del set-tore esamineranno le tendenze di mercato cheriguardano gli oli thailandesi e regionali, tra cui igrassi di palma, olio di soia; le prospettive di mer-cato per i biocarburanti, la domanda potenzialeglobale di olio di palma sostenibile, mercati diesportazione, e gli ultimi progressi tecnologici peraumentarne la produttività.Progetto di programma della conferenza:Sessione commerciale:- Supply and demand trends in the global and

ASEAN markets - Export marketsSessione tecnica:- Advances in technology and applicationsIl programma è soggetto a cambiamenti.Per informazioni contattare:Serena Lim, Editor, Oils & Fats International, UKTel: +44 1737 855066;E-mail: [email protected]://www.ofievents.com/asia/conference

Australasian Section AOCSBiennial Meeting & Workshops6-8 November 2013 - New South Wales, AustraliaAAOCS 2013 darà l'opportunità di conoscere ilprogresso dell'industria relativo gli aspetti riguar-danti gli oli e i grassi.Il tema di quest'anno concentra il dibattito sullaproduzione di oli per uso alimentare contro il loromaggiore uso di biocarburanti. Condurranno ladiscussione maggiori esperti delle industrie ali-mentari e di biocarburanti che esporranno la lorovisione in tema di produzione di olio e l'uso futu-ro. Sulla stessa linea, parteciperanno espertileader nella nuova area di biomateriali per discu-tere lo sviluppo della ricerca e come questoinfluenza il futuro degli oli commestibili e grassiindustriali.La gamma di argomenti in questo incontro prevede:• Food industry and Processing• New Analytical Methods• Agriculture • Aquaculture • Lipemic Index • Lipid Oxidation and Antioxidants • Omega-3 Fatty Acids • Fatty Acids, Lipids, and Health • Nutrition and Health • Lipids and Metabolic Syndrome • Lipids and Cognitive Function • Novel Foods and Supplements• Olive Oil and Other Vegetable Oils • BiotechnologyPer aggiornamenti consultare:http://www.aocs.org/Membership/content.cfm?ItemNumber=936

Notiziario• Olive Oil • Rapeseed Oil • Palm Oil • Sunflower Oil • Soybean Oil • Marine Oils, Micobial Lipids and Microalgae Oils • Oleochemistry, Biodiesel • SustainabilityPer aggiornamenti:http://www.eurofedlipid.org/meetings/antalya/index.htm

22nd IFSCC Conference 30 October-1 November 2013 - Rio de Janeiro, Brazil Annotare l’evento.Per informazioni e aggiornamenti: http://ifscc2013.com/en/index.php?pgID=informacoesdaconferencia&mi=010000000000

SCS Formulate 12-13 November 2013Annotare l’evento. Al momento non sono disponibi-li ulteriori informazioni.http://www.ifscc2013.com/

International Dairy Show 20133-6 November 2013 - Chicago, IllinoisLa fiera Dairy Show International torna in formatobiennale e propone temi sulla trasformazione lattie-ro-casearia, alimentare e delle bevande.Affronterà temi riguardanti la tecnologia, le innova-zioni nelle formulazioni, trasformazione, imballag-gio, distribuzione, sicurezza alimentare e altroancora.Un team di vendita e di marketing sarà presenteper valutare le tendenze, innovazioni e soluzioniper ridurre i costi e per fornire più valore ai clienti.http://www.dairyshow.com/

Practical Short Course on Trends inMargarine and Shortening Manufacture, Non-Trans Products03-07 November 2013 - Texas A&M, College Station,Texas, USAObiettivi del corso:Formare il personale in principi e pratiche di produ-zione di margarine a basso contenuto di grassi emargarine spalmabili trans-free• Tecniche di lavorazione • Garantire la qualità del prodotto• Dimostrazioni di prodotto utilizzando soia, semi dicotone, mais e oli di palmaPer iscrizioni e informazionihttp://foodprotein.tamu.edu/fatsoils/scmargarine.php

OFI Asia 20135-6 November 2013 - Bangkok, ThailandOFI Asia 2013 si terrà in Thailandia presso il Lan-dmark Hotel di Bangkok nelle giornate dal 5 al 6


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Notiz

iario mizzare la propria presenza in un unico evento

espositivo.Mostra convegno dedicata ai produttori di prodottichimici e naturali quali: - Additivi e Ausiliari- Polimeri e ResineSintetiche- Resine Naturali- Cariche e Riempitivi- Pigmenti- Coloranti e Tinture Naturali e Sintetiche- Fragranze- Essenze e Aromi- Oli e AcidiGrassi- Sostanze Naturali - Estratti e Prodotti Derivati- Prodotti Chimici di Base- Prodotti Chimici di Processo- Solventi e Diluenti Composti- Prodotti per il Trattamento delle Acque e altri vari

tipi di prodotti.Per l’utilizzo nei seguenti settori: - Coatings- Adesivi e Sigillanti- Inchiostri da Stampa- Plastica- Gomma- Compositi- Ceramica- Cosmetica- Detergenza- Vetro- Tessile e Tintoria- Calzaturificio- Cuoio- Conceria- Pelli Naturali e Sintetiche- Edilizia- Trattamento delle Superfici- Carta- Fonderia- Lubrificanti- Trattamento delle AcquePer informazioni:[email protected]

American Cleaning Institute AnnualMeeting & Industry ConventionJanuary 27-February 1, 2014 – Orlando, Florida, USALa Convention offre la possibilità di esaminareopportunità di business, strategie di marketing escambio di informazioni.Al momento non sono disponibili ulteriori informa-zioni. Per aggiornamenti:http://www.cleaninginstitute.org/about/acimeetings.aspx

Pittcon 2-7 March 2014 – Chicago, Illinois, USAConferenza dedicata a temi riguardanti la Chimicaanalitica e Spettroscopia applicata.http://www.labcanada.com/events/[email protected]

Annual National Institute of OilseedProducts (NIOP) Convention16-18 March 2014 - San Antonio, Texas, USAhttp://niop.org/

247th ACS National Meeting & Exposition16- 20 March 2014 - Dallas, Texashttp://portal.acs.org/portal/acs/corg/content

Ramspec - Rawmaterials, Specialties& CompoundsOttobre 2014 - MilanoLa mostra-convegno Ramspec si svolgerà a Mila-no con cadenza biennale e coinvolgerà 23/25 set-tori industriali utilizzatori di materie prime, prodottichimici e derivati. La prima edizione prevista a ottobre 2014, dedica-ta esclusivamente ai produttori di materie prime eprodotti chimici di processo, sarà un evento uniconel suo genere e permetterà alle industrie di otti-


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