Determination of cyanide compounds in kernels of stone-fruits Ingrid Steiner and Anatol Desser...

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Determination of cyanide compounds in Determination of cyanide compounds in kernels of stone-fruits kernels of stone-fruits Ingrid Steiner and Anatol Desser Vienna University of Technology, Institute of Chemical Engineering, Division of Natural Products and Food Chemistry, Getreidemarkt 9, A-1060 Vienna, Austria Introduction Kernels from stone-fruits can be used as a source of fat and protein, but their utilisation is connected with various technical and toxicological problems so that these kernels are normally burnt. One of these problems is the occurrence of cyanogenic compounds which is determined by the content of amygdalin. Amygdalin is decomposed by the enzyme emulsion inside the matrix into cyanogenic acid, benzaldehyde and glucose. Furthermore free cyanogenic acid is stored in cells of the stone. The Austrian food law and the EC regulation recommend 50 mg HCN / kg in foods (e.g. nougat and marzipan) corresponding to 846,1 mg amygdalin. Materials Kernels of apricots (Wachauer Marille from Austria harvested 2008) and sour cherries (from gardens in Vienna, Austria, harvested 2008). Methods Grinded kernels were mixed with buffer (pH = 5) and a yellow picrate paper was attached inside the vial without contact with the liquid phase. The vial was tightly closed and left standing at 30°C for 24 hours. In the presence of cyanide substances HCN is set free and the colour of the picrate paper changes from yellow to red according to the amount of HCN. After extraction of the paper with water the solution is analyzed by measuring the absorbance at 510 nm. Blank values and the calibration graph were determined in the same way. For this method it is important that the sample is grinded well because all cells and vacuoles have to be destroyed so that all cyanogenic glucosides can react with the enzymes. On the other hand the sample loses HCN while grinded which influences the results in a fatal way. The ideal time of grinding (=maximum release and minimum loss of cyanogenic glucosides) for 10 sour cherry kernels is approx. 120s. Correspondence to: Ao.Univ.Prof. Dr. Ingrid Steiner Institute of Chemical Engineering Getreidemarkt 9, A-1060 Vienna, Austria Tel: +43-1-58801/16002, Fax: +43-1-58801/16099 Email:[email protected] http://www.vt.tuwien.ac.at/ Results Conclusion The results show that it is not easy to find the best conditions for preparation of the kernels to analyze HCN contamination. So the next working steps should find out the optimal processes not only for apricot kernels but also for kernels of cherries, plums and peaches in correlation with the particle size. A possibility to accelerate the release of HCN from the samples might be the addition of immobilised enzymes. Storage of the kernels at higher temperatures results in lower residues of HCN so that it can be assumed that heat treatment and storage of grinded kernels increases the loss of HCN. Furthermore the residual amounts of HCN in grinded and original kernels after storage at different temperatures will be tested to check the possibility for use in food and feed industry under consideration of legal limits. Grinding conditions HCN (mg/kg) 30 s, interruptions 10 s 3100 ± 1233 30 s, without interruptions 2749 ± 955 40 s, interruptions 10 s 3517 ± 2775 40 s, without interruptions 2510 ± 961 50 s, without interruptions 2903 ± 759 120 s, without interruptions, charge 1 2764 ± 191 120 s, without interruptions, charge 2 1797 ± 257 120 s, without interruptions, charge 2 kernels stored 24 h at 30°C 1095 ± 214 Table 1. Kernels of sour cherries – Content of HCN equivalents for different grinding conditions Grinding conditions HCN (mg/kg) 20 s, without interruptions 159 ± 189 40 s, without interruptions 18,9 ± 4,3 60 s, without interruptions 3,3 ± 2,8 Table 2. Kernels of apricots - content of HCN equivalents for different grinding conditions 0 10 20 30 40 50 60 70 80 90 100 Cumulative frequency Q3 / % 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 Probabilit y density q3* 1 5 10 50 100 500 1000 Particle size [µm] Fig.2. Particle size of grinded (60 s) apricot kernels Acknowledgement We gratefully thank Dr.Gerd Mauschitz for measuring the particle size. 1. Formation of sodium isopurpurate after reaction of picric acid with HCN Fig.3. Decontamination of kernels Detoxification ofsour cherry kernels by treatm entwith ethanol 0 200 400 600 800 1000 1200 1400 1600 0 4 12 Time oftreatment[d] HCN [mg/kg] grinded kernels without treatment grinded kernels treated with EtOH whole kernels treated with EtOH „Wachauer Marille“ (apricot) Sour cherries

Transcript of Determination of cyanide compounds in kernels of stone-fruits Ingrid Steiner and Anatol Desser...

Page 1: Determination of cyanide compounds in kernels of stone-fruits Ingrid Steiner and Anatol Desser Vienna University of Technology, Institute of Chemical Engineering,

Determination of cyanide compounds in kernels of stone-fruitsDetermination of cyanide compounds in kernels of stone-fruitsDetermination of cyanide compounds in kernels of stone-fruitsDetermination of cyanide compounds in kernels of stone-fruits

Ingrid Steiner and Anatol Desser

Vienna University of Technology, Institute of Chemical Engineering, Division of Natural Products and Food Chemistry, Getreidemarkt 9, A-1060 Vienna, Austria

Introduction

Kernels from stone-fruits can be used as a source of fat and protein, but their utilisation is connected with various technical and toxicological problems so that these kernels are normally burnt. One of these problems is the occurrence of cyanogenic compounds which is determined by the content of amygdalin. Amygdalin is decomposed by the enzyme emulsion inside the matrix into cyanogenic acid, benzaldehyde and glucose. Furthermore free cyanogenic acid is stored in cells of the stone. The Austrian food law and the EC regulation recommend 50 mg HCN / kg in foods (e.g. nougat and marzipan) corresponding to 846,1 mg amygdalin.

Materials

Kernels of apricots (Wachauer Marille from Austria harvested 2008) and sour cherries (from gardens in Vienna, Austria, harvested 2008).

Methods

Grinded kernels were mixed with buffer (pH = 5) and a yellow picrate paper was attached inside the vial without contact with the liquid phase. The vial was tightly closed and left standing at 30°C for 24 hours. In the presence of cyanide substances HCN is set free and the colour of the picrate paper changes from yellow to red according to the amount of HCN. After extraction of the paper with water the solution is analyzed by measuring the absorbance at 510 nm. Blank values and the calibration graph were determined in the same way.For this method it is important that the sample is grinded well because all cells and vacuoles have to be destroyed so that all cyanogenic glucosides can react with the enzymes. On the other hand the sample loses HCN while grinded which influences the results in a fatal way. The ideal time of grinding (=maximum release and minimum loss of cyanogenic glucosides) for 10 sour cherry kernels is approx. 120s.

Correspondence to: Ao.Univ.Prof. Dr. Ingrid SteinerInstitute of Chemical EngineeringGetreidemarkt 9, A-1060 Vienna, AustriaTel: +43-1-58801/16002, Fax: +43-1-58801/16099Email:[email protected]://www.vt.tuwien.ac.at/

Results

ConclusionThe results show that it is not easy to find the best conditions for preparation of the kernels to analyze HCN contamination. So the next working steps should find out the optimal processes not only for apricot kernels but also for kernels of cherries, plums and peaches in correlation with the particle size. A possibility to accelerate the release of HCN from the samples might be the addition of immobilised enzymes.Storage of the kernels at higher temperatures results in lower residues of HCN so that it can be assumed that heat treatment and storage of grinded kernels increases the loss of HCN. Furthermore the residual amounts of HCN in grinded and original kernels after storage at different temperatures will be tested to check the possibility for use in food and feed industry under consideration of legal limits.

Grinding conditions HCN (mg/kg)

30 s, interruptions 10 s 3100 ± 1233

30 s, without interruptions 2749 ± 955

40 s, interruptions 10 s 3517 ± 2775

40 s, without interruptions 2510 ± 961

50 s, without interruptions 2903 ± 759

120 s, without interruptions, charge 1

2764 ± 191

120 s, without interruptions, charge 2

1797 ± 257

120 s, without interruptions, charge 2kernels stored 24 h at 30°C

1095 ± 214

Table 1. Kernels of sour cherries – Content of HCN equivalents for different grinding conditions

Grinding conditions HCN (mg/kg)

20 s, without interruptions 159 ± 189

40 s, without interruptions 18,9 ± 4,3

60 s, without interruptions 3,3 ± 2,8

Table 2. Kernels of apricots - content of HCN equivalents for different grinding conditions

0

10

20

30

40

50

60

70

80

90

100

Cumulative frequency Q3 / %

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

Probability density q3*

1 5 10 50 100

500 1000

Particle size [µm]

Fig.2. Particle size of grinded (60 s) apricot kernels

AcknowledgementWe gratefully thank Dr.Gerd Mauschitz for measuring the particle size.

Fig.1. Formation of sodium isopurpurate after reaction of picric acid with HCN

Fig.3. Decontamination of kernels

Detoxification of sour cherry kernels by treatment with ethanol

0

200

400

600

800

1000

1200

1400

1600

0 4 12

Time of treatment [d]

HCN

[mg/

kg]

grinded kernels withouttreatment

grinded kernels treated withEtOH

whole kernels treated withEtOH

„Wachauer Marille“ (apricot)

Sour cherries