UGANDA STANDARD
 
  US ISO 1208 
     First Edition
        2009-09-04
 
Reference numbe r
     US ISO 1208:1982
 © UNBS 2009
  
Spices and condiments — Determination of filth 
 US ISO 1208:1982 
 
© 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 
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Uganda National Bureau of Standards 
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Uganda 
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     US ISO 1208:1982  
© 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 1208, Spices and condiments — Determination of filth,  is identical with 
and has been reproduced from an International Standard, ISO 1208:1982,  Spices and condiments 
— Determination of filth, and adopted as a Uganda Standard.  
This standard was developed by the Food and agricu lture Standards Technical Committee (UNBS/TC 2).  
Wherever the words, "International Standard" a ppear, they should be replaced by "Uganda Standard." 
 International Standard 
INTERNATIONAL ORGANIZATION FOR STANDARDIZATION.ME)((CIYHAPO~HAF OPI-AHMJA~MR I-IO CTAH~APTM3AL&lMWIRGANISATION INTERNATIONALE DE NORMALISATION 
Spices and condiments - Determination of filth 
Epices - Dhtermination des impuretks 
First edition - 1982-12-15 
UDC 633.82/.84 : 543.869 Ref. No. ISO 12084982 (E) 
Descriptors : agricultural products, spices, Chemical analysis, determination of content, impurities. 
Price based on 5 pages Foreword 
ISO (the International Organization for Standardization) is a worldwide federation of 
national Standards institutes (ISO member bodies). The work of developing Inter- 
national Standards is carried out through ISO technical committees. Every member 
body interested in a subject for which a technical committee has been set up has the 
right to be represented on that committee. International organizations, governmental 
and non-governmental, in liaison with ISO, also take patt in the work. 
Draft International Standards adopted by the technical committees are circulated to 
the member bodies for approval before their acceptance as International Standards by 
the ISO Council. 
International Standard ISO 1208 was developed by Technical Committee ISO/TC 34, 
Agricultural food products, and was circulated to the member bodies in September 
1981. 
lt has been approved by the member bodies of the following countries: 
Australia 
Czechoslovakia 
Egypt, Arab Rep. of 
Ethopia 
France 
Germany, F. R. 
Hungary 
India 
Iraq 
The member bodies of 
on technical grounds : 
0 the following countries expressed disapproval of the document Israel 
Italy 
Kenya 
Malaysia 
Metherlands 
New Zealand 
Peru 
Philippines 
Poland Portugal 
Romania 
South Africa, Rep. sf 
Sri Lanka 
Tanzania 
Turkey 
United Kingdom 
Yugoslavia 
Canada 
USA 
International Organkation for Standardkation, 1982 l 
Printed in Switzerland INTERNATIONAL STANDARD ISO 12084982 (E) 
Spices and condiments - Determination of filth 
0 Introduction 
International Standards specifying requirements for spices and 
condiments prescribe, inter alia, that spices and condiments 
shall be practically free from dead insects, insect fragments and 
rodent contamination. For testing compliance with this require- 
ment, hand lens examination is adequate only in the case of 
whole spices and condiments. For ground spices and con- 
diments, the method to be used is that specified in this Interna- 
tional Standard, especially in cases of dispute. 
The method is applicable to most spices and condiments. In 
view of the number and variety of such products, however, it 
may be necessary in particular cases to modify the method or 
even to choose a more suitable method. Such modifications 
and other methods will be indicated in the International Stan- 
dards appropriate to the spices and condiments concerned. the washings for heavy filth and Sand. Washing the product 
with water, with or without treatment with pancreatin enzyme, 
and agitation with light Petroleum, the light filth collecting at 
the intet-face between the liquids after Separation. Transferente 
of the light filth to a filter Paper and microscopical examination 
for contaminants such as insect fragments and rodent hairs. 
5 Reagents 
The water used shall be distilled water or watet- of at least 
equivalent purity. 
5.1 Chloroform and, if required (sec 8.3), chloroform/ 
carbon tetrachloride mixtures. 
5.2 Pancreatin solution. 
1 Scope and field of application 
This International Standard specifies a method for the quan- 
titative determination of filth in spices and condiments. As no 
limits have been prescribed for filth in International Standards 
on spices and condiments, this method should be used for col- 
lecting more data and for settling international disputes. 
2 Reference 
ISO 948, Spices and condimen ts - Sampling. 
3 Definition 
For the purpose of this International Standard, the following 
definition applies. 
filth: Mineral matter (Sand, seil) and matter of animal origin (in- 
sect fragments, rodent hairs and excreta) separated from the 
product under the conditions specified. 
4 Principle 
Washing the product with chloroform (after, if necessary, 
preliminary extraction with light petroleum) and examination of Use pancreatin complying with the requirements of the annex, 
and which has been kept at about 10 OC. Use a Solution which 
has been freshly prepared as follows. 
Mix 10 g of pancreatin with 100 ml of warm water (temperature 
not exceeding 40 “CL Stir mechanically for 10 min or allow to 
stand for 30 min with intermittent stirring. Pour the Solution 
through a loosely packed pad of cotton wool, 100 mm thick, in 
a 60° funnel of diameter 100 to 125 mm. Repeat the filtration 
through the same pad. If filtration is slow in either case, filter 
with suction through a fast filter Paper using a Buchner funnel. 
If the filtration is still slow, pour the Solution through a slightly 
compressed cotton wool plug in the 60° funnel. Repeat if 
necessary, until the Solution filters rapidly through Paper. 
(Soluble pancreatin may be filtered directly through Paper with 
suction.) Dilute the filtrate to 100 ml for each 10 g Portion. 
5.3 Trisodium orthophosphate, 50 g/l Solution. 
5.4 Formaldehyde Solution. 
5.5 Light Petroleum, boiling range 40 to 60 0~. 
5.6 Light Petroleum, boiling range 100 to 120 OC. 
\ 1 ISO 1208-1982 (EI 
Apparatus 
Usual laboratory apparatus, and 
6.1 Trap flask (Wildman), consisting of a 1 000 ml conical 
flask into which is inserted a close-fitting rubber stopper carried 
on a rigid metal rod, 5 mm in diameter and approximately 
100 mm longer than the height of the flask. (A rod of a greater 
diameter is not desirable because of its greater displacement of 
liquid.9 The rod is threaded at its lower end and furnished with 
nuts and washers to hold it in place on the stopper. The lower 
nut and washer are countersunk in the rubber to prevent them 
from striking the flask. See the figure. 
Alternatively, a separating funnel, of capacity 1 000 ml, may be 
used. 
62 . 
63 . 
64 
ruled Beakers, of capacity 600 ml. 
Buchner funnel, of diameter 15 cm with a filter Paper. 
Buchner funnel, of diameter 7 cm with a filter Paper 
with parallel lines 5 mm apart. 
6.5 Filter Paper, ashless. 
6.6 Crucible, tared. 
67 m 
6.8 Oven, capable of being controlled at 80 OC. 
Wen, capable of being controlled at 103 OC. 
69 II 
6.10 Petri-dish, of diameter 80 mm. 
Suitable magnifying device (microscope, binocular Add 400 ml of the chIoroforn-~ (5.1) to the test Portion (8.1) or 
to the residue from the Operation described in 8.2. Allow the 
beaker to stand for at least 1 h with occasional stirring. 
Transfer the material and the solvent to the Buchner funnel 
(6.39, leaving the heavy residue sf sand and soij in the beaker. 
Drain. lf appreciable spiee tissue remains on the bottom of the 
beaker, add successive portions of cRIoroform mixed with car- 
bon tetrachloride to give increasingly higher relative densities 
until practically all spiee tissue has been removed by flotation. 
Transfer the heavy residue from the beaker to the ashless filter 
paper (6.5) and wash it with water to remove any sodium 
chloride present in the spiee. Examine this residue. If there is an 
appreciable residue, place the filter Paper in the tared crucible 
(6.6), ignite, and weigh the sand and soil. 
8.4 Treatment of re ue retained on the 
uchnes flmnel 
magnifier, etc. or preferably a wide-field stereoscopic micro- Dry the material retained on the Buchner funnel (See 8.3) for 
scope). 1 h in the oven (6.79, controlled at 80 OC. 
6.11 Balance. 
7 Sampling 
Sample the material by the method specified in ISO 848. 
8 Procedure 
NOTE - To achieve a satisfactory Separation of the light filth from 
spiee tissue, it may be necessary to remove most of the volatile oil and 
fat, or to treat the test Portion with pancreatin enzyme to digest starch 
and Protein, or both. 
If removal of volatile oil or fat is required, proceed as described in 8.2; 
otherwise, proceed directly as described in 8.3. 
8.1 Test Portion 8.5 Separation of light fikh 
Ensure that the test portio 
Sample (final lot Sample). n is represen tative of the laboratory Break the material into smakl pieces, weigh, to the nearest 
OS g, 25 g sf the Sample and transfer it to a breaker (6.2). 
8.1.2 Ground spices 
Waigh, to the nearest 0,l g, 25 g of the Sample and transfer it 
to a beaker (6.2). 
8.2 reliminâry remowal of ~~~~~~~ d dat 
Add 200 ml of the light Petroleum (5.5) to the test Portion (8.1) 
in the beaker. Boil gently on a warm water bath for 15 min. 
Remove the light Petroleum by decanting, taking care not to 
lose any of the material under test. 
8.3 Sepzwation of 
.‘& Procedure without enzyme treatment 
Transfer the residue to the trap flask (6.1). Add about 150 ml of 
water, heat to boiling and Simmer for 15 min with stirring. 
Wash down the inside of the flask with water and cool to below 
20 OC, diluting to about 600 ml with water. 
8.4.2 Paocedure with emyme treatment 
Transfer the dry residue to a beaker (6.2). Add 306 ml of water 
and stir until homogeneous. Add 50 ml of the pancreatin solu- 
tion (5.2) and mix. Adjust to pl-l 8 %tiith the trisodium ortho- 
Phosphate Solution (5.39. Readjust the pl-l after about 15 min, 
and again after about 45 min. Add 5 drops of the formaldehyde 
solution (5.4) and allow to digest overnight at 37 to 40 OC. Cool 
and transfer to the trap flask (6.i), making up to about 600 ml 
with water. 
8.51 Add, down the stirring rod of the flask, 25 ml of the 
light Petroleum (5.6). Tilt the trap flask at about 45O to the ver- 
2 ISO 12084982 (EI 
tical and mix for 1 min at the rate of 4 strokes per second with a 
brisk rotary motion so that the liquid is brought to a roll. Avoid 
splashing through the surface of the liquid in the trap flask with 
the rubber stopper. 
Allow to stand for 5 min. Then fill the flask 
allow to stand for 38 min. Stir every 5 min. with water and 
85.2 Spin the stopper to remove Sediment and raise the stop- 
per as far as possible into the neck of the flask, making sure 
that the light Petroleum layer and at least 1 cm of the liquid 
below the interface are above the stopper. Holding the stopper 
in place, transfer the trapped liquid to the Buchner funnel (6.4). 
Filter. 
8.5.3 Add 15 ml of the light Petroleum (5.6) to the contents 
of the trap flask and mix thoroughly. After 15 min, repeat the 
operations described in 8.52. If this second extraction yields an 
appreciable quantity of filth, decant most of the liquid from the 
flask, add another 15 ml of the light Petroleum (5.6) and carry 
out a third extraction. 
8.6 Wlicroscopical examination of light filth 
Remove the filter Paper from the Buchner funnel and place it on 
the Petri-dish (6.9). Place the Petri-dish in the oven (6.81, con- 
trolled at 103 OC, for 30 min. The dried filter Paper should be 
tightly Stuck to the Petri-dish. 
Examine the entire area of the filter Paper, using the magnifying 
device (6.10) in reflected light, scraping and probing with a 
mounted needle. 
Carry out the examination from left to right, from top to bottom 
for the first interval, from bottom to top for the second interval, 
from top to bottom for the third interval, etc. 9 Expression of resdts 
9.1 Heavy filth (sec 8.4) 
Note the presence of mineral matter. 
If the amount of sand and soil is such as to require calcination 
and weighing of these impurities, the mineral impurities con- 
tent of the product, expressed as a percentage by mass, is 
equal to 
100 
ml x - 
m0 
where 
m0 is the mass, in grams, of the test Portion; 
is the mass, in grams, of the residue obtained (8.3). 
9.2 Light filth (sec 8.6) 
Note the presence of matter of animal origin (see 8.6). 
If required, record separately the numbers of insect fragments, 
rodent hairs and other items of animal origin in the test Portion, 
i.e. the numbers per 25 g of Sample. 
10 Test report 
The test report shall show the method used and the resuits ob- 
tained. lt shall also mention any operating conditions not 
specified in this International Standard, or regarded as op- 
tional, as well as any circumstances that may have influenced 
the results. 
The test report shall include all the informa 
the complete identification of the Sample. tion necessary for ISO 1208-1982 (El 
Light Petroleum 
layer 
Water layer ,/” //A’ 
,-- Metal rod 1 
AA 
Rubber stopper _,-,-- Light Petroleum layer 
\ Nut (recessed into 
rubber stopper) 
Stopper lowered, for mixing, etc. Stopper raised, for trapping off Water layer 
Figure - Wildman trap flask (6.11, showing method of use 
NOTE - The upper nut and washer have been omitted. ISO 1208-1982 (EI 
Annex 
Pancreatin - 
AO General 
Pancreatin is a substance containing enzymes, principally pan- 
creatic amylase, trypsin, and pancreatic iipase, obtained from 
the pancreas of the hog, Sus scrofa Linne var. domesticus Gray 
(Farn. Suidae), or of the ox, Bos taurus Linne (Farn. Bovidae). 
Pancreatin converts not less than 25 times its mass of N.F.* 
Potato Starch Reference Standard into soluble carbo- 
hydrates, and not less than 25 times its mass of casein into pro- 
teoses. Pancreatin of a higher digestive power may be brought 
to this Standard by admixture with lactose, or with sucrose con- 
taining not more than 3,25 % of starch, or with pancreatin of 
lower digestive power. 
A.1 Description 
Pancreatin occurs as a cream-coioured, amorphous powder, 
having a faint, characteristic, but not offensive odour. Pan- 
creatin changes Protein into proteoses and derived substances, 
and converts starch into dextrins and sugars. Its greatest ac- 
tivities are in neutral or faintly alkaline media; more than traces 
of mineral acids or large amounts of alkali hydroxides render it 
inert. An excess of alkali carbonate inhibits its action. 
A.2 Assay for fat 
Introduce 2 g of pancreatin into a flask of capacity about 50 ml, 
add 20 ml of diethyi ether, stopper, and set it aside for several 
hours, mixing by rotating at frequent intervals. Decant the 
supernatant diethyl ether by means of a guiding rod into a plain 
filter Paper of diameter about 7 cm, previously moistened with 
diethyl ether, and collect the filtrate in a tared beaker. To the 
residue remaining in the flask, add a further 10 ml Portion of 
diethyl ether, proceeding as before, then add a third 10 ml por- 
tion of diethyl ether, and transfer the diethyl ether and the re- 
mainder of the pancreatin to the filter. Allow to drain, evapor- 
ate the diethyl ether spontaneously, and dry the residue at 
105 OC for 2 h: the residue of fat shall weigh not more than 
60 mg (3,O %). 
The fat may also be determined by the use of a Soxhlet 
continuous extraction apparatus. 
A.3 Assay for starch digestive power 
Determine the percentage of moisture in N.F. Potato Starch 
Reference Standard by drying about 500 mg of it, accurately 
weighed, at 120 OC for 4 h. Boil a sufficient quantity of water 
for 10 min, and cool to room temperature. Use this water for all 
subsequent dilutions specifying water. Specification 
Thoroughly mix a quantity of the N.F. Potato Starch Reference 
Standard, equivalent to 3,75 g of dry Reference Standard, with 
IO ml of water. Add the mixture, with constant stirring, 
to 75 ml of water, previously heated to about 55 OC, and con- 
tained in a tared 250 ml beaker. 
Rinse the remaining starch into the beaker with 10 ml of water. 
Heat the mixture to boiling, and boil it gently, with constant 
stirring, for 5 min. Add sufficient water to bring the mass of the 
mixture to 100 g, cool the Paste to 40 OC, and place the beaker 
in a water bath maintained at 40 OC. Suspend 150 mg of the 
pancreatin to be tested in 5 ml of water in a 250 ml beaker and 
add the Suspension to the starch Paste, mixing it weil by 
pouring the mixture from beaker to beaker for 30 s, noting the 
time when the pancreatin Suspension was first added to the 
Paste. Maintain the mixture at a temperature of 40 OC for 
exactly 5 min. Stir, at once add 0,l ml of this mixture to a 
solution previously made by adding 0,2 ml of iodine Solution, 
cwz 12) = 0,l mol / 1, to 60 ml of water, at a temperature of 23 
to 25 OC, and mix: no biue or violet colour shall be produced. 
A.4 Assay for casein digestive power 
Place 100 mg of finely powdered casein in a 50 ml volumetric 
fiask, add 30 ml of water, and Shake well to bring the casein 
into Suspension. Add 1,O ml of sodium hydroxide Solution, 
c(NaOH) = 0,l mol/l, and heat the mixture at 40 OC until the 
casein is completely dissolved, which should not require more 
than 30 min. Cool, dilute with water to 50 ml, and mix. 
Dissoive 100 mg of the pancreatin to be tested in 500 ml of 
water. Also dissoive 100 mg of N.F. Pancreatin Reference 
Standard for Casein Digestive Activity in another 500 ml of 
water. Mix 1 ml of glacial acetic acid with 9 ml of water and 
10 ml of ethanol. Place 5 ml of the casein Solution in each of 
two test tubes. To one of the test tubes add 2 ml of well- 
shaken pancreatin Solution and to the other test tube add 2 ml 
of the weli-shaken Reference Standard solution. Add 3 ml of 
water to each tube, mix by gentle agitation, immediately im- 
merse the test tubes in a water bath at 40 OC and keep at this 
temperature for 1 h. Then remove the tubes from the bath, and 
add 3 drops of the acetic acid mixture to each tube. 
The amount of turbidity in the test tube containing the Solution 
of pancreatin being tested shall not be greater than that in the 
test tube containing the Reference Standard soiution. 
A.5 Packaging and storage 
Preserve pancreatin in tight Containers, preferably at a tempera- 
ture of about 10 OC but in no circumstances above 30 OC. 
National Formulary (of USA). 
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