UGANDA STANDARD
 
        US ISO 5519
First Edition
2009-mm-dd
 
Reference numbe r  
    US ISO 5519:2008
 © UNBS 2009
  
Fruits, vegetables and derived products — Determination of 
sorbic acid content 
 US ISO 5519: 2008 
 
© UNBS  2009 – All rights reserved ii Compliance with this standard does not, of itself confer immunity from legal obligations 
A Uganda Standard does not purport to include all necessary provisions of a contract. Users are 
responsible for its correct application  
© UNBS 2009 
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilised in any 
form or by any means, electronic or mechanical, in cluding photocopying and microfilm, without prior written 
permission from UNBS. 
Requests for permission to reproduce this document should be addressed to 
The Executive Director 
Uganda National Bureau of Standards 
P.O. Box 6329 
Kampala  
Uganda 
Tel: 256 41 505 995 
Fax: 256 41 286 123 
E-mail: unbs@infocom.co.ug
Web: www.unbs.go.ug  
      US ISO 5519: 2008  
© UNBS  2009 - All rights rese rved                iii National foreword 
Uganda National Bureau of Standards (UNBS) is a parastatal under the Ministry  of Tourism, Trade and 
Industry established under Cap 327, of the Laws of U ganda. UNBS is mandated to co-ordinate the elaboration 
of standards and is  
(a) a member of International Organisation for Standardisation (ISO) and  
(b) a contact point for the WHO/FAO Codex A limentarius Commission on Food Standards, and  
(c) the National Enquiry Point on TBT/SPS Agr eements of the World Trade Organisation (WTO). 
The work of preparing Uganda Standards is carried out through Technical Committees. A Technical 
Committee is established to deliberate on standards in a gi ven field or area and consists of representatives of 
consumers, traders, academician s, manufacturers, governme nt and other stakeholders. 
Draft Uganda Standards adopted by the Technical Committ ee are widely circulated to stakeholders and the 
general public for comments. The committee reviews the comments before recommending the draft standards 
for approval and declaration as Uganda Standards by the National Standards Council. 
This Uganda Standard, US ISO 5519, Fruits, vegetables and derived products— Determination of sorbic acid
content,  is identical with and has been reproduced from an International Standard, ISO 5519:2008,
Fruits, vegetables and derived products— De termination of sorbic acid content, and adopted as a 
Uganda  Standard.  
This standard was developed under the subcommittee on fruits and vegetables by the Food and agriculture 
Standards Technical Committee (UNBS/TC 2). 
Wherever the words, "International Standard" a ppear, they should be replaced by "Uganda Standard." 
 INTERNATIONAL
STANDARDISO
5519
Second edition
2008-02-15
Reference number
ISO 5519:2008(E)
© ISO 2008Fruits, vegetables and derived 
products — Determination of sorbic acid content
Fruits, légumes et produits dérivés — Détermination de la teneur en 
acide sorbique
                                    
ISO 5519:2008(E)
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ISO 5519:2008(E)
© ISO 2008 – All rights reserved iiiContents  Page
1 Scope ........................................................................................................................ ............................  1
2 Principle .................................................................................................................... ............................  1
3 Reagents  .................................................................................................................... ..........................  1
4 Apparatus .................................................................................................................... .........................  2
5 Sample ....................................................................................................................... ...........................  3
6 Procedure .................................................................................................................... .........................  3
7 Calculation and expression of results ........................................................................................ .......  7
8 Repeatability ................................................................................................................ ........................  7
9 Test report .................................................................................................................. ..........................  8
Annex A (normative) Steam distillation apparatus ...................................................................................  9ISO 5519:2008(E)
iv © ISO 2008 – All rights reservedForeword
ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies
(ISO member bodies). The work of preparing International Standards is normally carried out through ISOtechnical committees. Each member body interested in a subject for which a technical committee has beenestablished has the right to be represented on that committee. International organizations, governmental and
non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International
Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.
International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. ISO shall not be held responsible for identifying any or all such patent rights.
ISO 5519 was prepared by Technical Committee ISO/TC 34, 
Food products , Subcommittee SC 3, Fruit and
vegetable products .
This second edition cancels and replaces the first edition (ISO 5519:1978), which has been technically revised.ISO 5519:2008(E)
© ISO 2008 – All rights reserved vIntroduction
The determination of the sorbic acid content of fruits, vegetables and derived products has been studied in
numerous projects during the acid's use as a fungicide, especially in wines. Because of its great volatility (verysimilar to that of acetic acid), the simplest extraction process is its entrainment by steam. This method has theadvantage of producing an almost pure aqueous solution of sorbic acid.
Two techniques for the determination of the quantity of sorbic acid contained in this solution are described in this
International Standard, namely:
— Technique A: spectrophotometry in the ultraviolet range, carried out after oxidation of sulfur dioxide, which
would interfere. The oxidation occurs spontaneously in a few minutes in air, after the addition of a trace of acopper catalyst.
— The natural essential oils of citrus fruits do not interfere with the determination, provided that they are
present in the small quantities normal in juice not enriched with essential oils. When the quantities ofessential oils are significant, they may be eliminated beforehand by the same method as that applied intechnique B.
— Technique B: colorimetry based on Schmidt's reaction, which requires the elimination of ethanol and
essential oils by the evaporation of an aliquot portion of the distillate. This technique, not as rapid as
technique A, but giving comparable results, is provided for use when a spectrophotometer allowing
measurements in the ultraviolet range is not available.
The interference caused by essential oils of garlic, onion or leek may be eliminated, when using either
technique, by the evaporation of an aliquot portion of the distillate..
viINTERNATIONAL STANDARD ISO 5519:2008(E)
© ISO 2008 – All rights reserved 1Fruits, vegetables and derived products — Determination of 
sorbic acid content
1S c o p e
This International Standard specifies a method for extracting the sorbic acid present in fruits, vegetables and
derived products, and two techniques for determining the sorbic acid extracted.
2 Principle
Homogenization of the product, followed by quantitative entrainment, by steam, of the sorbic acid present in atest portion. Determination of this acid in the distillate obtained, either by spectrophotometry in the ultravioletrange (technique A), or by measuring by photocolorimetry or by spectrophotometry, the pink colour obtainedafter oxidation by chromic acid and then treatment with thiobarbituric acid (technique B).
3R e a g e n t s  
Use only reagents of recognized analytical quality, and distilled water or water of at least equivalent purity.
3.1 Tartaric acid  [COOH (CH OH)2COOH], crystalline.
3.2 Sorbic acid  [CH3(CH:CH)2COOH],  standard solution, prepared by one of the following methods
(3.2.1 or 3.2.2).
3.2.1 Dissolve  of sorbic acid in  to  of a  sodium hydroxide solution. Transfer
quantitatively into a  volumetric flask, and dilute to the mark with water. Introduce  of the solution
obtained into a second  flask, and dilute to the mark with water.
3.2.2 Dissolve  of potassium sorbate, CH3(CH:CH)2COOK, (previously recrystallized and dried to
constant mass in a drier at  or in a desiccator over concentrated sulfuric acid) in water in a volumetric flask, and dilute to the mark with water. Introduce  of the solution obtained into a second
 flask, and dilute to the mark with water.
3.3 Calcium hydroxide  [Ca(OH)2] (if necessary), about  ( ) solution.
3.4 Acetic acid  (CH3COOH),  solution.
3.5 Lactic acid  [CH3CH(OH)COOH],  solution.
3.6 Copper , catalyst solution, for technique A.
In a  volumetric flask, dissolve, in a little water,  of sodium hydrogen carbonate (NaHCO3), and
 of pure copper(II) sulfate pentahydrate (CuSO4·5H2O). Dilute to the mark with water.
3.7 Chromic/sulfuric acid solution , for technique B.
Dissolve  of potassium dichromate (K2Cr2O7) in approximately  of water. Transfer quantitatively
into a  volumetric flask. Add  of a  sulfuric acid (H2SO4) solution. Dilute to the mark with
water.
1 litre of  sulfuric acid solution contains  of sulfuric acid, i.e.  of sulfuric acid, .0,010 g/l
0,100 g 10 ml 12 ml 0,1 N
1 000 ml 100 ml
1 000 ml
0,134 g
105◦C 1 000 ml
100 ml
1 000 ml
0,04 N 1,48 g/l
0,1 N
1N
1 000 ml 0,5 g
0,001 g
0,050 g 90 ml
200 ml 100 ml 0,3 N
0,3 N 14,7 g 8,4 ml ρ20=1,84 g/mlISO 5519:2008(E)
2 © ISO 2008 – All rights reserved3.8 Thiobarbituric acid solution , for technique B.
Dissolve  of thiobarbituric acid (C4H4N2O2S) in  of water to which has been added  of a 
sodium hydroxide (NaOH) solution. Transfer quantitatively into a  volumetric flask, and add  of a hydrochloric acid (HCl) solution. Dilute to the mark with water.
This solution is not stable and must be used within  following its preparation.
4 Apparatus
Usual laboratory apparatus and, in particular, the following.
4.1 Analytical balance .
4.2 Homogenizer or mortar , as appropriate.
4.3 Boiling water bath .
4.4 Steam distillation apparatus  (see Figure A.1), comprising the items listed in 4.4.1 to 4.4.5.
4.4.1 Steam generator flask , of capacity  to .
4.4.2 Bubbler , consisting of a cylindrical tube  in diameter and  in height, the lower part of
which is closed and enlarged into a sphere having a diameter of . The steam supply tube shall end
 above the bottom of the bubbler. The spherical part, in which the product is placed, may be heated either
electrically or by a flame. In the latter case, the burnt gases shall be deflected by a metal disc of diameter, having a central orifice of approximately  diameter in which the bottom of the bubbler isengaged. This device avoids the pyrogenation of materials which may be extracted from the product. The
auxiliary heating shall be controlled so that the volume of the product placed in the bubbler neither decreases
nor increases by more than  during the distillation.
4.4.3 Fractionating column , through which the vapour containing volatile acid passes. It may consist of
— a cylindrical tube of diameter  and height , containing a corrugated helix of No. 100 stainless
steel mesh, the helix having a pitch of , or
— a column of diameter  and height , having glass internal points, or— any other device having the same fractionating efficiency.
The fractionation of the vapour is indispensable to retain hydroxymethylfurfural when it is present. This
substance and its hydrolysis products absorb ultraviolet radiations at . The fractionating column may bereduced to  in height or be replaced by a Kjeldahl flask when the product is free ofhydroxymethylfurfural.
4.4.4 Condenser , of the West type, of effective length , placed vertically to ensure condensation of the
vapour and complete cooling of the distillate.
4.4.5 Receiver flasks , of capacities  or : 
— liquid products:  flask with a graduation mark at ;— thick or solid products:  flask.0,500 g 50 ml 10 ml 1N
100 ml 11 ml 1N
5h
1 000 ml 1 500 ml
30 mm 270 mm
60 mm
10 mm
150 mm
40 mm
5m l
20 mm 500 mm
15 mm
20 mm 600 mm
256 nm
200 mm
400 mm
200 ml 500 ml
200 ml 200 ml
500 mlISO 5519:2008(E)
© ISO 2008 – All rights reserved 34.4.6 Checking of efficiency of steam distillation apparatus .
The distillation apparatus shall allow  of distillate to be collected in  to , and shall also
comply with the following minimal conditions.
a) In normal distilling conditions,  of a known quantity of acetic acid added to the sample shall be found
in the distillate, which shall be . For this test, use  of a  acetic acid solution (3.4).
b) In the same distilling conditions, not more than 5 parts per thousand of a known quantity of lactic acid added
to the sample shall be found in the distillate, which shall be . For this test, use  of  lactic acidsolution (3.5).
4.5 Pipettes , of capacities ,  and . 
4.6 Graduated pipettes , of appropriate capacities.
4.7 Evaporating dish .
4.8 Conical flasks , for technique A, of capacity .
4.9 Spectrophotometer , for technique A, allowing measurements at a wavelength of  (ultraviolet),
with silica cells of  optical path length.
4.10 Volumetric flasks , for technique B, of capacity .
4.11 Photocolorimeter , fitted with a green filter, or spectrophotometer  allowing measurements at a
wavelength of , for technique B.
5S a m p l e
It is important that the laboratory receive a sample which is truly representative and has not been damaged orchanged during transport or storage.
6 Procedure
6.1 Preparation of test sample
6.1.1 Liquid products (juices, pulpy fluid products, syrups, etc.)
Thoroughly mix the laboratory sample.
6.1.2 Thick products (marmalades, jams, etc.)
Homogenize the laboratory sample after having carefully mixed it.
6.1.3 Solid products (fruits, vegetables)
Cut a part of the laboratory sample into small pieces, and remove seeds, stalks and carpellary cells, if
necessary, and carefully homogenize approximately  of the sample.
6.1.4 Frozen or deep-frozen products
After thawing the sample in a closed container and removing, if necessary, seeds, stalks and carpellary cells,
mix the product with the liquid formed during the thawing process and proceed as described in 6.1.1, 6.1.2 or6.1.3, as appropriate.300 ml 12 min 15 min
99,5 %
200 ml 20 ml 0,1 N
200 ml 20 ml 1N
10 ml 20 ml 25 ml
50 ml
256 nm
10 mm
25 ml
532 nm
40 gISO 5519:2008(E)
4 © ISO 2008 – All rights reserved6.2 Test portion
6.2.1 Liquid products
Using a pipette (4.5), take  of the test sample (6.1) and introduce it into the bubbler (4.4.2).The test portion may also be taken by mass, by weighing, to the nearest , approximately  of the test
sample.
6.2.2 Thick or solid products
Weigh, to the nearest , approximately  of the test sample (6.1) and introduce it into the bubbler (4.4.2)
with the minimum of water necessary to entrain the whole of the test portion and to make the mixture sufficientlyfluid.
NOTE In certain cases, it is necessary to leave the test portion to soak in the water for  to .
6.3 Distillation
6.3.1 General
Introduce  of the tartaric acid (3.1) into the bubbler (4.4.2) containing the test portion (6.2). Connect the
bubbler to the steam generator flask (4.4.1) and to the condenser (4.4.4) and simultaneously heat the flask andthe bubbler. Then carry out the distillation, making sure that the volume of the contents of the bubbler remainsconstant to within .
6.3.2 In the case of liquid products (6.2.1)
Collect the distillate in the  receiver flask (4.4.5), stopping the distillation when the  mark is
reached.
6.3.3 In the case of thick or solid products (6.2.2)
Collect, in the  flask (4.4.5), a volume of distillate at least 20 times greater than the volume of the
contents of the bubbler. Measure the volume ( ) collected, using a graduated cylinder.
6.4 Technique A: determination by spectroph otometry in the ultraviolet range
6.4.1 Determination
6.4.1.1 If the initial product contains essential oils of garlic, onion or leek, the presence of these essential oils
causes significant absorbance, especially in the case of garlic. Complete evaporation1) of the distillate, after its
being made alkaline, allows the effect of this absorbance to be counteracted.
When these essential oils are present, therefore, take, with a pipette (4.5),  distillate (6.3) and transfer it to
a small evaporating dish (4.7); make it alkaline with  to  of the calcium hydroxide solution (3.3),evaporate it to dryness on the boiling water bath (4.3) and reconstitute it with water to re-establish the initialvolume.
1) The evaporation to dryness does not destroy the sorbic acid if the conditions are sufficiently alkaline.10 ml
0,01 g 10 g
0,01 g 10 g
1h 2h
0,5 g
5m l
200 ml 200 ml
500 ml
V
25 ml
1,5 ml 2m lISO 5519:2008(E)
© ISO 2008 – All rights reserved 56.4.1.2 As appropriate, take with a pipette (4.5)  (see Note) of the distillate (6.3) or of the reconstituted
solution (6.4.1.1), place it in a  conical flask (4.8), and add  of the copper catalyst solution (3.6).
Shake briefly and leave to stand in contact with air for several minutes.
NOTE The volume of  is intended for products containing up to  of sorbic acid per litre or per kilogram. For
higher contents, take only  or  and dilute to  with water.
Measure the absorbance of the solution using the spectrophotometer (4.9), at a wavelength of .
Subtract, from the value found, the absorbance of the blank test solution (6.4.2).
6.4.2 Blank test
Carry out a blank test in parallel with the determination, replacing the  of distillate by  of water.
6.4.3 Number of determinations
Carry out two determinations on the same test sample (6.1).
6.4.4 Preparation of calibration curve
6.4.4.1 Into a series of six  conical flasks (4.8), introduce respectively, with a graduated pipette (4.6):
; ; ; ;  and  of the standard sorbic acid solution (3.2); make up the volume to  by
adding water.
The solutions obtained contain: ; ; ; ;  and  of sorbic acid per litre.6.4.4.2 To each flask, add  of the copper catalyst solution (3.6).
Measure the absorbances of the solution using the spectrophotometer (4.9) at a wavelength of .
Subtract, from the values found, the absorbance of the blank test solution (6.4.2).6.4.4.3 Plot the calibration curve, showing the absorbances of the solution (6.4.4.2) as a function of the sorbic
acid concentrations of the solutions obtained in 6.4.4.1, i.e. before addition of the copper catalyst solution (3.6),expressed in milligrams per litre.
6.5 Technique B: determination by photocol orimetry or by spectrophotometry at 
6.5.1 Determination
6.5.1.1 If the initial product contains ethanol, remove it from the distillate by the following method.
Using the pipette (4.5) introduce  of the distillate (6.3) into a small evaporating dish (4.7), and make it
alkaline with  to  of the calcium hydroxide solution (3.3), place the dish on the boiling water bath (4.3),and evaporate until the volume is reduced by about half, which normally takes about . Quantitativelytransfer the residue into the  volumetric flask (4.10). Make up to the mark with rinsing water from the dish.Shake.
6.5.1.2 If the initial product contains essential oils (in the case of juice from citrus fruits), eliminate them from
the distillate by the same method as described in 6.5.1.1, but prolong the evaporation so that a volume of 
to  is attained.
6.5.1.3 If the initial product contains essential oils of garlic, onion or leek, proceed as indicated in 6.4.1.1.
NOTE In the case of garlic, even when the distillate is evaporated to dryness, a very weak absorbance, corresponding to
 of sorbic acid per kilogram, remains.10 ml
50 ml 10 ml
10 ml 200 mg
5m l 2m l 10 ml
256 nm
10 ml 10 ml
50 ml
0m l 1m l 2m l 3m l 5m l 10 ml 10 ml
0m g 1m g 2m g 3m g 5m g 10 mg
10 ml
256 nml
532 nm
25 ml
1,5 ml 2m l
30 min
25 ml
1m l
2m l
1,5 mgISO 5519:2008(E)
6 © ISO 2008 – All rights reserved6.5.1.4 As appropriate, take, with a pipette (4.5),  (see next paragraph) of the distillate (6.3) or of the
reconstituted solution obtained after treatment (6.5.1.1, 6.5.1.2, or 6.5.1.3), and introduce it into the 
volumetric flask (4.10).
The volume of  is intended for products containing up to  of sorbic acid per litre or per kilogram. For
higher contents, take only  or  and dilute to  with water.
Add  of the chromic/sulfuric acid solution (3.7) and keep the flask for  in the boiling water bath (4.3).
Add  of the thiobarbituric acid solution (3.8) and keep the flask for  in the boiling water bath for a
further ; a pink colour will develop. Cool in an iced water bath and dilute to the mark with water.
Within , measure the absorbance of the solution using the photocolorimeter or spectrophotometer (4.11)
at a wavelength of .
Subtract, from the value found, the absorbance of the blank test solution (6.5.2).
6.5.2 Blank test
Carry out a blank test in parallel with the determination, replacing the  of distillate by  of water.
6.5.3 Number of determinations
Carry out two determinations on the same test sample (6.1).
6.5.4 Preparation of calibration curve
6.5.4.1 Prepare a  standard sorbic acid solution by diluting 1 volume of the standard solution (3.2) with
4 volumes of water.
6.5.4.2 Into a series of six  volumetric flasks (4.10), introduce respectively, with a graduated pipette (4.6):
; ; ; ;  and  of the diluted standard sorbic acid solution (6.5.4.1); make up the volume
to  by adding water.
The solutions obtained contain: ; ; ; ;  and  of sorbic acid per litre.
6.5.4.3 To each flask add  of the chromic/sulfuric acid solution (3.7) and keep the flasks for  in the
boiling water bath (4.3).
Add  of the thiobarbituric acid solution (3.8) and keep the flasks for  in the boiling water bath for a
further ; a pink colour will develop. Cool in an iced water bath and dilute to the mark with water.
Within , measure the absorbance of the solution using the photocolorimeter or spectrophotometer (4.11)
at a wavelength of .
Subtract, from the value found, the absorbance of the blank test solution (6.5.2).
6.5.4.4 Plot the calibration curve, showing the absorbance of the solutions (6.5.4.3) as a function of the
corresponding concentration of sorbic acid, expressed in milligrams per litre.10 ml
25 ml
10 ml 200 mg
5m l 2m l 10 ml
4m l 10 min
4m l 10 min
20 min
30 min
532 nm
10 ml 10 ml
2,0 mg/l
25 ml
0m l 2m l 4m l 6m l 8m l 10 ml
10 ml
0m g 0,4 mg 0,8 mg 1,2 mg 1,6 mg 2,0 mg
4m l 10 min
4m l 10 min
20 min
30 min
532 nmISO 5519:2008(E)
© ISO 2008 – All rights reserved 77 Calculation and expression of results2)
7.1 Test portion measured by volume
The sorbic acid concentration, expressed in milligrams per litre of product, is given by the formula
where
is the mass of sorbic acid, expressed in milligrams, per litre of distillate (6.5.1), read on the
calibration curve (see 6.4.4 or 6.5.4);
is the volume, in millilitres, taken in 6.4.1.2 or 6.5.1.4 (usually , but may be reduced to  or
).
7.2 Test portion measured by mass
The sorbic acid content, expressed in milligrams per kilogram of product, is given by the formula
where
 is the mass, in grams, of the test portion (6.2.2); 
is the mass of sorbic acid, expressed in milligrams, per litre of distillate (6.3.3), read on the
calibration curve (see 6.4.4 or 6.5.4);
is the volume, in millilitres, of distillate collected (see 6.3.3);is the volume, in millilitres, taken in 6.4.1.2 or 6.5.1.4 (usually , but may be reduced to  or
).
8 Repeatability
The absolute difference between two single test results, obtained with the same method on identical test
material in the same laboratory by the same operator using the same equipment within a short interval of time,will in not more than  of cases be greater than  the arithmetic mean of the two results.
2) Various vegetable products contain small quantities of volatile substances which may be extracted by organic solvents
and which absorb  radiation or give the coloured reaction used in technique B (6.5). Therefore, results which are
only slightly positive (less than  per litre or per kilogram) should be interpreted with caution and compared with results
from the same products free from sorbic acid.256 nm
10 mgm1×200
V1
m1
V1 10 ml 5m l
2m l
m1×V×10
m0×V1
m0
m1
V
V1 10 ml 5m l
2m l
5% 5%ISO 5519:2008(E)
8 © ISO 2008 – All rights reserved9 Test report
The test report shall specify:
a) all information necessary for the complete identification of the sample;b) the sampling method used, if known;
c) the test method used,
 with reference to this International Standard;
d) all operating details not specified in this International Standard, or regarded as optional, together with
details of any incidents which may have influenced the test result(s);
e) the test result(s) obtained, or, if the repeatability has been checked, the final quoted result obtained.ISO 5519:2008(E)
© ISO 2008 – All rights reserved 9Annex A
(normative)
Steam distillation apparatus
Key
1 steam generator flask (4.4.1)
2 bubbler (4.4.2)
3 fractionating column (4.4.3)
4 condenser (4.4.4)
5 receiver flask (4.4.5)
Figure A.1 — Diagram of steam distillation apparatus (4.4)ISO 5519:2008(E)
ICS 67.080.01
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© ISO 2008 – All rights reserved