NEW DEVELOPMENT FOR PASTA-PRODUCTS: IMMATURE WHEAT AND FUNCTIONAL COMPONENTS NEW DEVELOPMENT FOR...

NEW DEVELOPMENT FOR NEW DEVELOPMENT FOR PASTA-PRODUCTS: PASTA-PRODUCTS:

IMMATURE WHEAT AND IMMATURE WHEAT AND FUNCTIONAL COMPONENTSFUNCTIONAL COMPONENTS

NEW DEVELOPMENT FOR NEW DEVELOPMENT FOR PASTA-PRODUCTS: PASTA-PRODUCTS:

IMMATURE WHEAT AND IMMATURE WHEAT AND FUNCTIONAL COMPONENTSFUNCTIONAL COMPONENTS

III PASTA WORLD CONGRESS, 23III PASTA WORLD CONGRESS, 23rdrd – 26 – 26thth October 2005 October 2005III PASTA WORLD CONGRESS, 23III PASTA WORLD CONGRESS, 23rdrd – 26 – 26thth October 2005 October 2005

** CRA – Istituto Sper. Cerealicoltura CRA – Istituto Sper. CerealicolturaRoma Roma

** CRA – Istituto Sper. Cerealicoltura CRA – Istituto Sper. CerealicolturaRoma Roma

M. G. D’EGIDIOM. G. D’EGIDIO*, *, M. A. PAGANIM. A. PAGANI**, M. C. CASIRAGHI**, **, M. C. CASIRAGHI**, M. ZARDIM. ZARDI**** and C. CECCHINI* and C. CECCHINI*

M. G. D’EGIDIOM. G. D’EGIDIO*, *, M. A. PAGANIM. A. PAGANI**, M. C. CASIRAGHI**, **, M. C. CASIRAGHI**, M. ZARDIM. ZARDI**** and C. CECCHINI* and C. CECCHINI*

**** Università degli Studi Università degli Studi Milano Milano

**** Università degli Studi Università degli Studi Milano Milano

FOODPROPERTIES

in the past

nowadays

NO ADVERSE EFFECTS

• WELL-BEING• BETTER HEALTH• REDUCTION OF THE RISK OF DISEASES

FUNCTIONAL FOOD

FUNCTIONAL FOOD

“A food can be regarded as functional if it is satisfactorily demonstrated to affect one or more target functions in the body beyond adequate nutritional effects, in a way that is relevant either to improve the state of health and well-being and/or the reduction of risk of disease” (ILSI, 2002)

III PASTA WORLD CONGRESS, 23III PASTA WORLD CONGRESS, 23rdrd – 26 – 26thth October 2005 October 2005III PASTA WORLD CONGRESS, 23III PASTA WORLD CONGRESS, 23rdrd – 26 – 26thth October 2005 October 2005

D’ Egidio et al. New development for pasta products:immature wheat and functional components

FUNCTIONAL COMPONENTSFUNCTIONAL COMPONENTS

FOS FRUCTOLIGOSACCHARIDES

carbohydrates forming a short linear chain of fructose (2-9 units) with a single glucose unit at the head

present as a plant storage carbohydrates in a number of vegetables and plants including wheat and other cereals, onion, bananas, garlic and chicory

soluble in water

non digestible in the small intestine (1.5 kcal/g)

typically non viscous

highly fermentable

PREBIOTIC EFFECTPREBIOTIC EFFECT

n

Mitsuoka et al.,1987; Tashiro et al., 1992; Spiegel et al., 1994

III PASTA WORLD CONGRESS, 23III PASTA WORLD CONGRESS, 23rdrd – 26 – 26thth October 2005 October 2005


0

5

10

15

20

25

30

35

0 5 10 15 20 25 30 35 40

days after anthesis

FOS (

%d.m

.)

0

10

20

30

40

50

60

70

80

mois

ture

(%

)

cv OFANTO durum wheat

cv PANDAS common wheat

Studies have shown that a large quantity of FOS is stored in the stems and grains of WHEAT for the first period of its growing cycle.

IMMATURE WHEAT at “MILKY PHASE”

9 DAA 13 DAA 17 DAA 21 DAA 28 DAA 35 DAA

days a

fter

an

thesis

(D

AA

)

durum wheat is richer in FOS than common wheat

moisture

MILKY PHASEMILKY PHASE WAXY PHASEWAXY PHASE PHISIOLOGICAL PHASEPHISIOLOGICAL PHASE



drying at 35°C 24- 48 h

EARS at MILKY PHASEMOISTURE 65%

MOISTURE KERNELS 13-15%

MILLING

WHOLE MEAL FLOUR

IMMATURE WHEAT GRAINS (IWG) need a drying process in order to reduce their moisture content and so to be used as raw material in

conventional processes

THRESHING

LABORATORY SCALE

The whole process is to be optimized for industrial

scale



CHARACTERISTICS OF IMMATURE WHEAT GRAINS (IWG)CHARACTERISTICS OF IMMATURE WHEAT GRAINS (IWG)

more balanced aminoacid composition (4-5 g lys /100 g protein)

IWG: natural source of FOS

lower level of starch (40-50%) and amylose (~20%)

higher content of soluble sugars (2-5%)

more relevant -amylase activity (2-15 IU/g d.m)similar protein content but higher level of albumins and globulins and lower/absence of gliadins and glutenins

incapability of forming GLUTEN NETWORK

higher content of total fiber (13-15%)

Evolution of main components in durum wheat grains



0

10

20

30

40

50

60

5 10 15 20 25 30

days after anthesis

% w

.b

0

0.5

1

1.5

2

2.5

3

3.5

4

4.5

5

g ly

sine

/100

g pr

otei

n

FOS

protein content

total starchlysine

soluble sugarsg

g

MILKY PHASE

necessity to use IWGIWG flour in mixture with semolina in conventional pasta-

making process

in comparison with MATURE WHEAT GRAINS, IWG present a:

AIM IAIM I

characterization of protein and starch fractions of IWG in order to identify the more suitable

technological processes to obtain FUNCTIONAL FOODS

PROTEIN

Protein solubility Accessibility of thiols SDS-PAGE characterization IMMUNOBLOTTING

characterization Surface hydrophobicity

STARCH

Total starch content Amylose/Amilopectine

ratio Soluble Sugars Viscoamylographic test Microscopic investigation



PROTEIN EVOLUTION DURING RIPENING (cv OFANTO)PROTEIN EVOLUTION DURING RIPENING (cv OFANTO)

PROTEINS ACQUIRE A MORE COMPACT AGGREGATION STATE AFTER THE MILKY PHASE

the decrease in albumins and globulins is associated to a noticeable increase in protein aggregates requiring urea and DTT for their solubilizationa marked decrease of both total and accessible thiol is assorbed at 17 daa

0

0.5

1

1.5

2

2.5

3

3.5

5 10 15 20 25 30

days after-anthesis

mic

rom

ol th

iol/g fl

our

total thiol

accessible thiol

17th day

0

20

40

60

80

5 10 15 20 25 30 35

days after anthesis

mg

prot

eins

/g fl

our

soluble proteins

insoluble proteins

urea and DTT soluble proteins

17th day



IWG could be an interesting raw material for

GLUTEN FREE FOOD

characterization by SDS-PAGE and IMMUNOBLOTTING using commercial anti-gliadin antibodies confirmed that synthesis

of the major gluten polypeptides begins at 17 days after anthesis, and that no immunoreactive gluten material was

present at 9 or 13 daa

SDS-PAGESDS-PAGE

GLIA

DIN

GLIA

DIN

28282121171799 1313

days after anthesisdays after anthesis9 13 17 21 28

GLIA

DIN

days after anthesisdays after anthesis

IMMUNOBLOTTINGIMMUNOBLOTTING

kDakDa

9494

6868

4545

2929

2020



STARCH PROPERTIES in IMMATURE and MATURE grainsSTARCH PROPERTIES in IMMATURE and MATURE grains

IMMATURE GRAINS

MATURE GRAINS

IMMATURE GRAINS MATURE GRAINS

Ø granules = 20-30 µm Ø granules = 2-3 µm

Characteristics of starch can influence its

digestibility

IWG: potential ingredient for FUNCTIONAL FOOD

IWGflour

semolina

STARCH (%d.b.) 59.4 79.0

AMYLOSE/AMYLOPECTINE

20.3/79.7

23.3/76.4

SOLUBLE SUGARS (%w.b.)

5.6 1.7

-AMYLASE ACTIVITY (UI/g d.b.)

7.5 0.2

TEMPERATURE CONDITIONS



study of the characteristics of IWG – enriched foods

AIM IIAIM II

DRIED PASTA

FRESH PASTA BREAD



EXPERIMENTAL PASTA-PRODUCTSEXPERIMENTAL PASTA-PRODUCTS

DRIED PASTA: SPAGHETTI SHAPE ( = 1.65mm)

• 30% IWG and 70% semolina

Formulation:

• 100% semolina (REFERENCE SAMPLE)

HT

LT

PILOT PLANT SCALE (CRA-ISC, Rome)

HIGH TEMPERATURE: Tmax = 90°C; 8h

LOW TEMPERATURE: T = 50°C; 20 h



100% semolina 30% IWG +70% semolina

COLOUR DETERMINATIONCOLOUR DETERMINATIONCOLOUR DETERMINATIONCOLOUR DETERMINATION

0

5

10

15

20

25

30

35

40

pasta products

yell

ow

in

dex

HTLT

0

10

20

30

40

50

60

70

pasta products

bro

wn

in

dex

LTHT

furosine level= 755 mg /100g protein

more than 50% LYSINE UNAVAILABLE

100% semolina 30% IWG

30% IWG +70% semolina100% semolina

CHEMICAL COMPOSITION OF DRIED PASTA*CHEMICAL COMPOSITION OF DRIED PASTA*

100% semolina

SPAGHETTI TOTAL FIBRESOLUBLE SUGARS

30% IWG + 70% semolina

PROTEINTOTAL STARC

H

FOS

11.1

11.8

74.9

67.4

0.70

0.90

3.1

5.2

1.7

4.0



RESISTENT STARCH

1.1

1.5

Exp

eri

men

tal

** values reported on packaging

14.6**

13.0**

59.0**

63.5**

1.63

2.38

4.6

3.0

15.1

6.9

dietetic pasta for diabetics

(with INULIN)

100% wholemeal semolina

4.5

3.6

Com

merc

ial

* data expressed as % db; all analyses in duplicate



Cooking behaviour of pastaCooking behaviour of pasta

Leaching of FOS after cookingbefore cooking

after cooking

DISADVANTAGE OF DRIED PASTA

DECREASE OF BENEFITS APPORTED BY

ENRICHEMENT WITH IWG

DISPITE THE LEACHING OF FOS INTO THE COOKING WATER,

ENRICHED PASTA HAS A HIGHER FOS CONTENT THAN CORRESPONDING

REFERENCE SAMPLE

EVALUATION OF COOKING QUALITY

65WHOLEMEAL PASTA

56DIETETIC PASTA

6930% IWG HT

69100% sem HT

6330% IWG LT

61100% sem LT

TOTAL SCORE *SAMPLE

OPTIMAL COOKING TIME (min)

8.30

8.00

11.30

8.30

8.00

8.30* TOTAL SCORE 10 low quality –100 high quality

according to D’Egidio et al., 1993

FIRMNESS STICKINESS BULKINESS

65

88

68

73

60

73

60

20

70

68

60

55

70

60

70

65

70

55

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

FO

S (

g/8

0g

pa

sta

po

rtio

n)

30% IWG LTDIETETIC PASTA

“IN

VIT

RO

” S

TA

RC

H D

IGESTIO

N“I

N V

ITR

O ”

STA

RC

H D

IGES

TIO

Nacc

ord

ing t

o B

righenti

et

al.,

19

95

COOKED SPAGHETTI at optimal cooking time

SIMULATION OF CHEWINGportion of food containg 2g of available carbohydratesmixed with 5ml 20 mM

sodium phosphate buffer (pH 6.9) containing 10 mm NaCl and 25 U of human salivary amylase gram of

starch

SIMULATION OF STOMACH DIGESTIONacidification with HCl 8 M pH 2.0

digestion at 37°C with hog pepsine per gram of starch 1 h

5 min

SIMULATION OF SMALL INTESTINE DIGESTIONcorrection of pH from 2.0 to 6.9 with NaOH 8 M

mixture was placed in DIALYSIS BAGS with hog pancreatine 50 U per gram of starch and to simulate peristalsis a glass ball was inserted into

the tube and used to mix the content of the tube every 15 min

5 hbags were suspended in separate sealed containers filled with 450 ml of

phosphate buffer at 37°C every 30 min 1 ml sample of dialysate was taken for analysisof total dialysable reducing sugars by the 3,5-dinitrosalicylic acid method

TIME OF ANALYSIS

STEPS



“IN VITRO STARCH DIGESTION”“IN VITRO STARCH DIGESTION”

0.0

5.0

10.0

15.0

20.0

25.0

30.0

35.0

40.0

45.0

0 30 60 90 120 150 180 210 240 270 300 330time (min)

% d

iges

ted

star

ch

EFFECT OF IWG ADDITIONEFFECT OF IWG ADDITION

100% semolina LT

30% IWG LT

GROWING AWARENESS FOR FOOD WITH LOW STARCH DIGESTIBILITY



0.0

5.0

10.0

15.0

20.0

25.0

30.0

35.0

40.0

45.0

0 30 60 90 120 150 180 210 240 270 300 330

time (min)

% d

iges

ted

star

ch

100% semolina LT

30% IWG LT

WHOLEMEAL PASTA

PASTA for diabetics

COMPARISON WITH COMMERCIAL PRODUCTSCOMPARISON WITH COMMERCIAL PRODUCTS

Pasta enriched with IWG was characterized

by a lower starch availability than the corrisponding 100%

semolina

Pasta enriched with IWG had a % of available

starch lower than the product for diabetics



Glycemix IndexGlycemix IndexThe Glycemic Index (GI) is defined as the incremental area under blood glucose response curve (AUC) elicited by a portion of food containing 50 g available carbohydrate expressed as the percent of AUC elicited by 50 g ofglucose taken by the same subjects.

METHODMETHOD

0;15;30;45;60;90;120 min: finger prick blood samples(glucose determination: YSI

2000 STAT Glucose Analyser)

7 meals:

3 standard meals (500 ml water + 50 g

glucose)

4 spaghetti meals

N° subjects = 10 (5 men; 5 women)

RangeMean ± semCharacteristics

Age (years)

Weight (kg)

Height (m)

BMI (kg/m2)

Basal glycemia (mg/dl)

25,8 ± 0,5

66.4 ± 4.39

1.74 ± 0,03

21,7 ± 0,8

75,7 ± 8,8

20,1 - 26,6

22 - 28

51 - 87

1,60 - 1,87

63 - 90

NO DIFFERENCES IN GLYCEMIC INDEX AMONG PRODUCTS



0.0

10.0

20.0

30.0

40.0

50.0

60.0

70.0

0 15 30 45 60 75 90 105 120

time (min)

bloo

d gl

ucos

e (m

g/dl

)

glucose 100% semolina 30% IWG inuline wholemeal

GLYCEMIC INDEXGLYCEMIC INDEXPOST-PRANDIAL GLUCOSE RESPONSEPOST-PRANDIAL GLUCOSE RESPONSE

The post-prandial glucose response was similar among pasta meals, showing a peak value at 30 min and a return to baseline values within 2 hours, with no significant differences in blood glucose concentration at any given time points. The calculated GI values of experimental pasta resulted slightly higher but not significantly different than those evaluated for commercial products.

similar post-prandial blood glucose responses among pasta meals

67 6453 58

100

0

20

40

60

80

100

120

glucose 100%semolina

30% IWG dieteticpasta

wholemeal

CONCLUSIONSCONCLUSIONS

DRIED PASTA ENRICHED WITH IWG

ACCEPTABLE COOKING QUALITY

PRODUCT OF MEDIUM-LOW GLICEMIC INDEX

HIGH FOS CONTENT BUT LEACHING OF THESE MOLECULES DURING COOKING



FRESH PASTA ENRICHED WITH IWG

(short cooking time)

REDUCTION OF FOS LEACHING DURING COOKING

WORK IN PROGRESSWORK IN PROGRESS

DRIED AND FRESH PASTA ENRICHED WITH

PRE- COOKED IWG FLOUR

REDUCTION OF STARCH DIGESTIBILITY PASTA WITH LOW GLYCEMIC INDEX



Gabriella Bottega

Carolina Palandi

Chiara Balansino

Stefania Iametti

Franco Bonomi

Thank you!

“IN

VIT

RO

” S

TA

RC

H D

IGESTIO

N“I

N V

ITR

O ”

STA

RC

H D

IGES

TIO

Nacc

ord

ing t

o B

righenti

et

al.,

19

95

COOKED SPAGHETTI at optimal cooking time

SIMULATION OF CHEWINGportion of food containg 2g of available carbohydratesmixed with 5ml 20 mM

sodium phosphate buffer (pH 6.9) containing 10 mm NaCl and 25 U of human salivary amylase gram of

starch

SIMULATION OF STOMACH DIGESTIONacidification with HCl 8 M pH 2.0

digestion at 37°C with hog pepsine per gram of starch 1 h

5 min

SIMULATION OF SMALL INTESTINE DIGESTIONcorrection of pH from 2.0 to 6.9 with NaOH 8 M

mixture was placed in DIALYSIS BAGS with hog pancreatine 50 U per gram of starch and to simulate peristalsis a glass ball was inserted into

the tube and used to mix the content of the tube every 15 min

5 hbags were suspended in separate sealed containers filled with 450 ml of

phosphate buffer at 37°C every 30 min 1 ml sample of dialysate was taken for analysisof total dialysable reducing sugars by the 3,5-dinitrosalicylic acid method

The amount of released reducing sugars (consisting of dialysed fragments of digested starch plus the native reducing sugars) was

expressed in maltose equivalent and as percentage of total available carbohydrates after adjustment for the rate of sugars

diffusion

RESULTS

TIME OF ANALYSIS

STEPS



FOS DETERMINATION

FOS DETERMINATION

PRELIMINARY EXTRACTION WITH ETHANOL 95%

1 h at 80°C

WATER EXTRACTION

2 h at 105°C

FRUCTOSE CONTENTGLUCOSE CONTENT

ENZYMATIC PROCEDURE (GOPOD) CHEMICAL PROCEDURE (resorcine-HCl)

ethanolic extract

FRUCTAN

FRUCTOSE CONTENT

GLUCOSE CONTENT

ENZYMATIC PROCEDURE (GOPOD)

CHEMICAL PROCEDURE (resorcine-HCl)

SOLUBLE SUGARSresidual

according to D’Egidio et al., 1993

enzymatic attach

III PASTA WORLD CONGRESS, 23rd – 26th October 2005


FOS DETERMINATION

FOS DETERMINATION

SAMPLE

boiling water 10 min at 85°C

EXTRACTION DISSOLUTION

sugar analysis 1

HYDROLYSIS

amyloglucosidase 30 min at 60°C

sugar analysis 2

HYDROLYSIS

fructozyme 30 min at 60°C

sugar analysis 3

AOAC 1995 Revision March 1998

where K= conversion factorGlucose= from inulin = glucose from Hydrolisis II - Glucose from Hydrolisis I- Glucose in free sugar - glucose from disaccharides in free sugar (saccarose and maltose).Fructose= from inulin = fructose from Hydrolisis II - fructose in free sugar - fructose from disaccharides in free sugar (saccarose)

Inulin= K (Glucose + Fructose)



FRESH PASTA : TAGLIATELLE SHAPE

• 30% IWG and 70% semolina

Formulation:

• 100% semolina (REFERENCE SAMPLE)

100% semolina 30% IWG +70% semolina

moisture = 30%

sfoglia thickness = 2mm

2 CYCLES OF PASTEURIZATION 95°C 1 min before packaging100°C 45 min after packaging

LABORATORY PLANT SCALE

PASTA- PRODUCTSPASTA- PRODUCTS

DOUGH


FRESH PASTA CHARACTERIZATIONFRESH PASTA CHARACTERIZATION

DRIED PASTA FRESH PASTA 0

10

20

30

40

50

60

70

pasta products

bro

wn

in

dex

BROWN INDEX

30% IWG

100% semolina

0

1

2

3

4

5

100%sem 30% IWG

FOS

(%

d.b

.)

before cooking

after cooking

100% SEMOLINA 30% IWG

Leaching of FOS after cooking

PROCESSESCumulative

volume(mm3 /g)

POROSITY

(%)

COOKING TIME (min)

Pressure-extrudedpast

a26.95 3.75 8 –11

Role-sheeted pasta

51.22 6.94 1 -2

the more porose the structure, the more available the starch

30 mg /100g protein

112 mg /100g protein

FUROSINE LEVEL

HIGHER % OF DIGESTED STARCH THAN DRIED PASTA

0.0

10.0

20.0

30.0

40.0

50.0

60.0

70.0

80.0

90.0

100.0

0 30 60 90 120 150 180 210 240 270 300 330

time (min)

% d

iges

ted s

trac

h

100% semolina

30% IWG

30% IWG LT

“IN VITRO “STARCH DIGESTION

100% semolina LT



02468

101214161820

5 10 15 20 25 30

days after anthesis

% w

.b. TOTAL PROTEIN

NEW DEVELOPMENT FOR PASTA-PRODUCTS: IMMATURE WHEAT AND FUNCTIONAL COMPONENTS NEW DEVELOPMENT FOR...

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