IS 2557 (B) (1994): Annatto colour for food products ...2 vkj tgk¡] R = Na vFkok K iii vkbZ,l@IS 2557 : 1994 Structural Formula lhvks ,p Molecular Weight — Bixin 394.50 Norbixin

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है”ह”ह

IS 2557 (B) (1994): Annatto colour for food products –Specification [FAD 8: Food Additives]

© BIS 2012

B U R E A U O F I N D I A N S T A N D A R D SMANAK BHAVAN, 9 BAHADUR SHAH ZAFAR MARG

NEW DELHI 110002

Qjojh@February 2012 ewY; oxZ@Price Group 2

vkbZ,l@IS 2557 : 1994¼f}Hkk"kh@Bilingual½

(iquiqZ"V@Reaffirmed 2004)

Hkkjrh; ekud

[kk| mRiknksa osQ fy, vÂkVks jax — fof'kf"V¼ igyk iqujh{k.k ½

¼la'kksèku la[;k 1 vkSj 2 lekfo"V½

Indian Standard

ANNATTO COLOUR FOR FOODPRODUCTS — SPECIFICATION

( First Revision )(Incorporating Amendments No. 1 and 2)

;wMhlh@UDC 633.863.92 : 637.2/.3

© Hkk ek C;wjks 2012

Hkkjrh; ekud C;wjksekud Hkou] 9 cgknqj 'kkg t+Qj ekxZ

ubZ fnYyh 110002

[kk| la;ksth foHkkxh; lfefr] ,iQ,Mh 8

izkDdFku

bl Hkkjrh; ekud (igyk iqujh{k.k) osQ elkSns dks [kk| la;ksth foHkkxh; lfefr }kjk vafre :i fn, tkus rFkk [kk| vkSj Ñf"kfoHkkx ifj"kn }kjk vuqeksfnr fd, tkus ds ckn Hkkjrh; ekud C;wjks }kjk xzg.k fd;k x;kA

vUukVks jax ,fjy (cht dh dksfVax) ftls okf.kfT;d Hkk"kk esa lkekU; :i ls fcDlk vksjhysuk&,y- ds ikSèks ds cht ds :iesa ekuk tkrk gS] ls izkIr fd;k tkrk gSA rsy esa bldk ?kksy eD[ku vkSj vU; [kk| inkFkksZa dks jax nsus ds fy, iz;ksx fd;ktkrk gS tcfd ikuh esa bldk ?kksy vkSj mlesa FkksM+k lk vYdkyh feyk dj iuhj vkSj vU; leku izdkj ds mRiknksa dks jaxnsus ds fy, fd;k tkrk gSA

;g ekud izkd`frd [kk| jaxksa dh Hkkjrh; ekudksa dh Ük`aa[kykvksa esa ls ,d gS ftldh vuqefr LokLF; ea=kky;] Hkkjr ljdkj}kjk tkjh [kk| vifeJ.k fuokj.k fu;e] 1955 rFkk mlds uohure la'kksèkuksa }kjk nh xbZ gSA bl ekud dks rS;kj djus ds fy,bu fu;eksa ij fo'ks"k è;ku fn;k x;k gSA rkSy ,oa eki ekudksa (fMCckcan oLrq,a) fu;e] 1977 ij Hkh fo'ks"k è;ku fn;k x;kgSA rFkkfi] ;g ekud bu fu;eksa ds varxZr yxk, x, izfrcaèk dh 'krZ tgka Hkh ykxw gks] ij ykxw gksrk gSA

;g ekud lcls igys 1963 esa izdkf'kr fd;k x;k FkkA ns'k esa izkS|ksfxdh; fodkl dks è;ku esa j[krs gq, rFkk varjkZ"Vªh;fof'kf"V;ksa ds cjkcj ykus osQ fy, bldk iqujh{k.k fd;k x;k gSA bl iqujh{k.k esa dSjVhuk;M~l dh U;wure lhek,a rFkk Hkkjhèkkrqvksa dh vfèkdre rFkk vUukVks ds fy, igpku ijh{k.kksa dks Hkh 'kkfey fd;k x;k gSA

Food Additives Sectional Committee, FAD 8

FOREWORD

This Indian Standard (First Revision) was adopted by the Bureau of Indian Standards, after the draft finalized bythe Food Additives Sectional Committee had been approved by the Food and Agriculture Division Council.

Annatto colour is obtained from the aril (coating of the seed) commonly known in commerce as the seed of theplant Bixa orellana L. Its solution in oil is used for colouring butter and other food products while its solution inwater with a little alkali is used for colouring cheese and other similar products.

This standard is one of a series of Indian Standards on natural food colours permitted under the Prevention ofFood Adulteration Rules, 1955 with the latest amendments, issued by the Ministry of Health, Government ofIndia. In the preparation of this standard due consideration has been given to these Rules. Due consideration hasalso been given to the Standards of Weights and Measures (Packaged Commodities) Rules, 1977. However, thisstandard is subject to the restriction imposed under these Rules wherever applicable.

This standard was first published in 1963. It has been revised taking into account the latest technologicaldevelopments in the country and also to bring it in line with the International specifications. In this revisionminimum limits for carotenoids and maximum limits for heavy metals have been included as well as identificationtests for annatto.

i

vkbZ,l@IS 2557 : 1994

bl ekud dks rS;kj djus ds fy, fuEufyf[kr izdk'kuksa ls Hkh lgk;rk yh xbZ gSA

[kk| la;ksft;ksa dh igpku o 'kq¼rk ds fy, fof'kf"V] [kk| ,oa ikSf"Vdrk isij] 19] la;qDr jk"Vª la?k rFkk fo'o LokLF;laxBu dh [kk| ,oa d`f"k laxBu }kjk izdkf'krA

iQwM dsehdy dksMsDl] foKku vkSj jk"Vªh; vuqlaèkku ifj"kn dh jk"Vªh; vdkneh] okf'kaxVu Mhlh] la;qDr jkT; vejhdk]}kjk izdkf'krA 1981 ds laLdj.k dk f}rh; ifjf'k"VA

fooj.kfooj.kfooj.kfooj.kfooj.k

Js.kh & dkWjVsukbZMldksM la[;k & lhvkbZ (1975) la[;k 75120

lhvkbZ (1975) izkÑfrd larjk 4

bZbZlh la[;k bZ&160 ch

jklk;fud uke

rsy esa vUukVks rÙo ds cgqr ls jaxhu la?kVd gksrs gSaA buesa ls ,d izeq[k fcfDlu gS tks lhvkbZ,l rFkk Vªkal nksuksa izdkjksa esagks ldrk gSA fcfDlu ds FkeZy MhxzsMs'ku izkWMDV~l Hkh fo|eku gks ldrs gSaaA

ikuh esa ?kqyu'khy vUukVks esa ukSjfcfDlu rÙo fufgr gksrk gSA ;g fcfDlu dk gkbMªksfyfll mRikn gS tks lksfM;e ;k iksVsf'k;elkYV ds :i esa gksrk gS vkSj ;g jaxus (dyfjax) dk izeq[k fl¼kar gSA lhvkbZ,l vkSj Vªkal nksuksa izdkj blesa fo|eku gks ldrs gSaA

jklk;fud iQkewZyk µ fcfDlu lh25,p30vks4ukWjfcfDlu lh24,p28vks4

In the preparation of this standard, assistance has been derived from the following publications.

Specification for Identity and Purity of Food Additives, Food and Nutrition Paper 19, published by Food andAgriculture Organization of the United Nations and the World Health Organization.

Food Chemical Codex published by the National Academy of Sciences and National Research Council,Washington DC, USA. Second supplement of 1981 Edition.

Description

Class — Carotenoids

Code Number — CI (1975) No. 75120CI (1975) Natural Orange 4EEC No. E-160 b

Chemical Name

Annatto extract in oil contains several coloured components, the major single one being bixin, which may bepresent in both Cis and trans forms. Thermal degradation products of bixin may also be present.

Water soluble annatto contains norbixin the hydrolysis product of bixin, in the form of sodium or potassiumsalt, as the major colouring principle. Both Cis and trans forms may be present.

Chemical Formula — Bixin C25

H30

O4

Norbixin C24

H28

O4

vkbZ,l@IS 2557 : 1994

ii

lajpukRed iQkewZyk lhvkbZ,l vkSj Vªkal fcfDludh lajpuk

,p3lhvks2lh lhvks

2,p

,p3lhvks

2lh

lhvkbZ,l vkSj Vªkal ukWjfcfDlu(,u, vFkok ds lkYV~l)

dh lajpuk

vkjvks2lh lhvks2vkj

vkjvks2lh lhvks2vkj

tgk¡] R = Na vFkok K

iii

vkbZ,l@IS 2557 : 1994

Structural Formula

lhvks2,p

Molecular Weight — Bixin 394.50

Norbixin 380.48

For the purpose of deciding whether a particular requirement of this standard is complied with, the final value,observed or calculated, expressing the result of a test or analysis, shall be rounded off in accordance with IS 2 : 1960‘Rules for rounding off numerical values (revised)’. The number of significant places retained in the rounded offvalue should be the same as that of the specified value in this standard.

vk.kfod Hkkj µ fcfDlu 394-50ukWjfcfDlu 380-48

;g fuf'pr djus osQ fy, fd bl ekud esa fdlh vis{kk fo'ks"k dk ikyu fd;k x;k gS ;k ugha] rks ijh{k.k ;k fo'ys"k.k esaifj.kke dks n'kkZus okys voyksdu ;k x.kuk }kjk izkIr vafre eku dks vkbZ,l 2 % 1960 ^la[;kRed ekuksa osQ iw.kk±du lacaèkhfu;e (iqujhf{kr)* osQ vuqlkj iw.kk±fdr dj fn;k tk,A iw.kk±fdr eku esa j[ks x, lkFkZd LFkkuksa dh la[;k mruh gh gks ftruhbl ekud esa fufnZ"V eku dh gSA

1

vkbZ,l@IS 2557 : 1994

Hkkjrh; ekud

[kk| mRiknksa osQ fy, vÂkVks jax — fof'kf"V¼ igyk iqujh{k.k ½

Indian Standard

ANNATTO COLOUR FOR FOODPRODUCTS — SPECIFICATION

( First Revision )

1 SCOPE

1.1 This standard prescribes the requirements and themethods of test for annatto colour derived from theplant Bixa orellana L. and supplied in the form of(a) solution in oil, and (b) solution in water, for use inbutter, cheese and other food products.

2 REFERENCES

The Indian Standards listed at Annex A are necessaryadjuncts to this standard.

3 TYPES

3.1 The material shall be of the following two types:

a) Solution in oil for use in butter and other foodproducts, and

b) Solution in water for use in cheese and otherfood products.

4 REQUIREMENTS

4.1 General

The material shall be derived only from the plant Bixaorellana L. and shall not contain any extraneouscolouring matter. It shall be processed, packed, storedand distributed under hygienic conditions in licencedpremises (see IS 2491 : 1972).

4.2 Solution of Annatto Colour in Oil for Use inButter and Other Food Products

4.2.1 Annatto extract in oil, as solution or suspension,is prepared by extraction of the outer coating of seedswith vegetable oils. In the preparation of the solutionof annatto colour in oil, only the vegetable oilsincluded under the Prevention of Food AdulterationRules, 1955 shall be used, either singly or in a mixture

1 fo"k;1 fo"k;1 fo"k;1 fo"k;1 fo"k; {k s=k{k s=k{k s=k{k s=k{k s=k

1-1 1-1 1-1 1-1 1-1 ;g ekud eD[ku] iuhj vkSj vU; [kk| inkFkksZa esa iz;ksxfd, tkus okys vUukVks jax tks fd fcDlk vksjhysuk&,y- ikSèksls izkIr dj (d) rsy esa ?kksy] vkSj ([k) ikuh esa ?kksydjmiyCèk djk;k tkrk gS] dh vko';drkvksa vkSj ijh{k.k dhizfØ;kvksa dk fuèkkZj.k djrk gSA

2 lanHkZ2 lanHkZ2 lanHkZ2 lanHkZ2 lanHkZ

vuqcaèk d esa n'kkZ, x, Hkkjrh; ekud bl ekud ds vko';dlgk;d gSaA

3 izdkj3 izdkj3 izdkj3 izdkj3 izdkj

3-13-13-13-13-1 lkexzh fuEufyf[kr nks izdkj dh gksaxh%

d) eD[ku vkSj vU; [kk| inkFkksZa esa iz;ksx ds fy, rsyesa ?kksy_ vkSj

[k) iuhj vkSj vU; [kk| inkFkksZa esa iz;ksx ds fy, ikuhesa ?kksyA

4 vis{kk,¡4 vis{kk,¡4 vis{kk,¡4 vis{kk,¡4 vis{kk,¡

4-14-14-14-14-1 lkekU; lkekU; lkekU; lkekU; lkekU;

lkexzh vko';d :i ls dsoy fcDlk vksjhysuk&,y- ikSèks lsgh yh tkuh pkfg, vkSj dksbZ Hkh vU; vfrfjDr jaxus okyhlkexzh ugha gksuh pkfg,A bls ykblsal okys {ks=kksa esa (ns[ksavkbZ,l 2491 % 1972) rS;kj djuk] iSd djuk] HkaMkj.kdjuk vkSj LoPN ifjfLFkfr;ksa esa forfjr djuk gksxkA

4-2 eD[ku vkSj vU; [kk| inkFkks Z a esa iz;ksx ds4-2 eD[ku vkSj vU; [kk| inkFkks Z a esa iz;ksx ds4-2 eD[ku vkSj vU; [kk| inkFkks Z a esa iz;ksx ds4-2 eD[ku vkSj vU; [kk| inkFkks Z a esa iz;ksx ds4-2 eD[ku vkSj vU; [kk| inkFkks Z a esa iz;ksx dsfy, rsy esa vUukVks jax dk ?kksyfy, rsy esa vUukVks jax dk ?kksyfy, rsy esa vUukVks jax dk ?kksyfy, rsy esa vUukVks jax dk ?kksyfy, rsy esa vUukVks jax dk ?kksy

4-2-14-2-14-2-14-2-14-2-1 rsy esa ?kksy fuyacu ds :i esa vUukVks vdZ chtksa dsfNydksa dks ouLifr rsy dh lgk;rk ls fudkydj rS;kjfd;k tkrk gSA rsy esa vUukVks jax dk ?kksy rS;kj djus ds fy,dsoy [kk| vifeJ.k fuokj.k fu;e] 1955 esa 'kkfeyouLifr rsy dk iz;ksx vdsys vFkok feJ.k ds :i esa fd;k

2

vkbZ,l@IS 2557 : 1994

and shall conform to relevant Indian Standard whereit exists.

4.2.2 The solution of annatto colour in oils shall beclear and shall remain so on storage in suitablecontainers at 15°C except for a slight deposit ofstearine or shall be in the form of a suspension. Thesuspension on dilution with hot oil to bring the bixincontent to 0.24 percent shall be a clear solution.

4.2.3 Colour

The colour of solution in amyl acetate (seeIS 231 : 1957) at a dilution of 1 : 1 000 (w/v), whenmeasured in a Lovibond Tintometer with a 1-cm cell,shall be not less than the following:

Yellow units 5.0

Red units 0.4

or be not less than the colour of the following inorganicsolution at a liquid depth of one centimetre which maybe employed for matching the stated dilution in aplunger type colorimeter using incident light closelyapproximating the north day light:

Potassium bichromate 0.320 g

Cobalt ammonium sulphate 2.02 g

[CoSO4, (NH

4)

2 SO

4, 6H

2O]

Sulphuric acid sp-gr 1.84 2 ml

Distilled water To make solutionto one litre

These reagents shall be of the analytical reagent grade(see IS 250 : 1964 and IS 266 : 1993). Although thesolution retains its tinctorial value for a considerabletime, after prolonged storage, its optical clarity shallbe examined before use, to ensure that no alterationhas taken place.

NOTES

1 Diluted solution of annatto colour in amyl acetate is not stablein colour quality particularly if exposed to light, andmeasurement should be carried out on the diluted solutionwithout undue delay.

2 In using the Tintometer, the maker’s instructions shall be rigidlyfollowed.

4.3 Solution of Annatto Colour in Water for Use inCheese and Other Food Products

4.3.1 Water soluble annatto is prepared by extractionof the outer coating of the seeds with aqueous alkali(sodium or potassium hydroxide). In the preparationof the solution, potable water shall be used. A littlequantity (0.5 to 3 percent) of alkali may be added (seeIS 376 : 1986 and IS 296 : 1986).

tk, vkSj og lacafèkr Hkkjrh; ekud] tgka dgha gks] dsvuq:i gksA

4-2-24-2-24-2-24-2-24-2-2 rsy esa vUukVks jax dk ?kksy mi;qDr fMCcksa esa 15°lsa-ij HkaMkj esa lkiQ jguk pkfg, flok; FkksM+s fLVvfju dsvykok ;k lLisa'ku ds :i esa gksuk pkfg,A fcfDlu va'k dks0-24 izfr'kr rd ykus ds fy, xje rsy ds lkFk ?kksyus ij;g ?kksy lkiQ gksuk pkfg,A

4-2-34-2-34-2-34-2-34-2-3 jax

1 % 1 000 (MCY;w@oh)] ij ?kksyus ij ,fey ,flVsV esa ?kksyds jax (vkbZ,l 231 % 1957 ns[ksa) dks tc 1&lseh- lSy dslkFk yksch ckW.M fVaVksehVj esa ekik tk, rks og fuEufyf[krls de ugha gksuk pkfg,%

ihyh bdkb;ka 5-0yky bdkb;ka 0-4

;k mldk jax fuEufyf[kr buvkxsZfud ?kksy ds 1 lseh- dhnzoh; xgjkbZ ds jax ls de ugha gksuk pkfg,A bldk mi;ksxIyaxj Vkbi dyjhehVj esa cuk, x, jax dk eSfpax djus dsfy, fnu ds izdk'k esa mi;ksx fd;k tk ldrk gS%

iksVsf'k;e ckbØksesV 0-320 xzkedksckYV veksfu;e lYiQsV 2-02 xzke¹lhvks,lvks

4] (,u,p

4)2 ,lvks

4]

6,p2vksº

lyÝ;wfjd ,flM ,lih&thvkj 1-84 2 feyh-fMfLVYM okWVj ,d yhVj ?kksy

cukus ds fy,

;s jhtsaV fo'ys"k.kkRed jhtsaV xzsM ds gksaxs (vkbZ,l 250 %1964 vkSj vkbZ,l 266 % 1993 ns[ksa)A ;|fi ?kksy viuhewy ekU;rk dkiQh le; rd cuk, j[krk gS ysfdu iz;ksxdjus ls igys mls dkiQh le; rd mldh DysfjVh dh tkapdh tkuh pkfg, rkfd ;g lqfuf'pr fd;k tk lds fd mlesadksbZ ifjorZu ugha gqvk gSA

fVIif.k;ka

1 1 1 1 1 ,fey ,lhVsV esa vUukVks jax dk ikuh Myk gqvk ?kksy vius jax dhxq.koÙkk dks ugha cuk, j[k ldrk] fo'ks"kdj rc tcfd mls izdk'kesa j[kk tk,A ?kksy dks fcuk nsjh fd, eki fy;k tkuk pkfg,A

2 2 2 2 2 fVaVksehVj dk iz;ksx djrs le; mlds fuekZrk osQ funsZ'kksa dk l[rhls ikyu fd;k tkuk pkfg,A

4-3 iuhj vkSj vU; [kk| inkFkk s ± es a ikuh es a4-3 iuhj vkSj vU; [kk| inkFkk s ± es a ikuh es a4-3 iuhj vkSj vU; [kk| inkFkk s ± es a ikuh es a4-3 iuhj vkSj vU; [kk| inkFkk s ± es a ikuh es a4-3 iuhj vkSj vU; [kk| inkFkk s ± es a ikuh es avUukVks jax ds ?kksy dk iz;ksxvUukVks jax ds ?kksy dk iz;ksxvUukVks jax ds ?kksy dk iz;ksxvUukVks jax ds ?kksy dk iz;ksxvUukVks jax ds ?kksy dk iz;ksx

4-3-1 4-3-1 4-3-1 4-3-1 4-3-1 ikuh esa ?kqyu'khy vUukVks chtksa dh ckgjh dksfVax dks,Doh;l vYdyh (lksfM;e ;k iksVsf'k;e gkbMªksDlkbZM) }kjkfupksM+dj rS;kj fd;k tkrk gSA bl ?kksy dks rS;kj djrs le;ihus ;ksX; ikuh dk mi;ksx djuk pkfg,A vYdyh dh FkksM+hek=kk (0-5 ls 3 izfr'kr) Mkyh tk, (vkbZ,l 376 % 1986vkSj vkbZ,l 296 % 1986 ns[ksa)A

3

vkbZ,l@IS 2557 : 1994

4.3.2 The solution shall be clear and shall remain soon storage in suitable containers at a temperature of15°C.

4.3.3 Colour

The colour of the solution in 0.1 N sodium hydroxideof potassium hydroxide at a dilution 1 : 1 000 (m/v)measured in a 1-cm cell shall be the same as thatspecified in 4.2.3.

4.4 Identification

4.4.1 Spectrophotometry

Annatto extracts in oil, diluted with chloroform exhibitsabsorbance maxima at 439, 470 and 501 nm. Watersoluble annatto extracts, diluted with water exhibitabsorbance maxima at 453 and 482 nm.

4.4.2 Column Chromatography and Carr-PriceReaction

4.4.2.1 Preparation of alumina column

Prepare a small chromatograph column by filling a glasstube (7 × 200 mm internal diameter), stoppered withglass wool, with alumina (800 – 200 mesh) slurried inbenzene so that the settled alumina fills about two-thirdsof the tube. Fill the column with benzene and adjustthe flow rate to about 30 drops/min using a rubber outlettube and clamp until the benzene layer is 2-3 mm abovethe alumina surface.

NOTE — It is advisable to place a glass-wool plug at the top ofthe alumina to prevent churning when solvent is added, and toapply a reservoir of solvent at the top of the column, which willexert a mild pressure on the chromatographic column. Thesuction must be carefully controlled to prevent channelling inthe alumina. Avoid eluting the benzene or other solvent completely.It is necessary that the surface of the alumina is kept covered withliquid layer.

4.4.2.2 Annatto extract in oil

Add to the top of the alumina column 3 ml of a solutioncontaining sufficient sample, in benzene, to impart acolour equivalent to a 0.1% potassium dichromatesolution; elute with benzene until a pale yellow fractionis washed from the column. Wash the column with three10-ml volumes of dry chloroform; then add 5 ml ofCarr-Price reagent and allow it to run onto the top ofthe column. The orange-red zone (bixin) at the top ofthe column immediately becomes blue-green.

4.4.2.3 Water soluble annatto

Transfer 2 ml or 2 g to a 50-ml separatory funnel andadd sufficient 2 N sulphuric acid to make the solution

4-3-2 4-3-2 4-3-2 4-3-2 4-3-2 ;g ?kksy 15°lsa- rkieku esa mi;qDr fMCcksa esa lkiQjguk pkfg, vkSj ;g fLFkfr HkaMkj.k esa Hkh cuh jguhpkfg,A

4-3-34-3-34-3-34-3-34-3-3 jax

?kksy dk jax 1 lseh- lSy esa ekik x;k 0-1 ,u lksfM;egkbMªksDlkbM dks 1 % 1 000 (,e@oh) ij ikuh Mkydjiryk djus ij mldk jax mlh izdkj gksuk pkfg, tSlkfd 4-2-34-2-34-2-34-2-34-2-3 esa fofufnZ"V fd;k x;k gSA

4-4 igpku4-4 igpku4-4 igpku4-4 igpku4-4 igpku

4-4-14-4-14-4-14-4-14-4-1 LisDVªksiQksVksehVjh

rksy esa vUukVks vdZ 439] 470 vkSj 501 ,u,e ij DyksjksiQkeZbthfcVl ,clkjcsal eSfDlek ij ?kqy tkrk gSA ikuh esa?kqyu'khy vUukVks vdZ 453 vkSj 482 ,u,e ij okWVjbXthfcV ,cjlkjcsal eSfDlek ij ?kqy tkrk gSA

4-4-2 4-4-2 4-4-2 4-4-2 4-4-2 dkWye ØksesVksxzkiQh rFkk dkj&izkbl dh izfrfØ;k

4-4-2-1 4-4-2-1 4-4-2-1 4-4-2-1 4-4-2-1 ,ywfeuk dkWye rS;kj djuk

,d dkap dh V~;wc (7 × 200 feeh- van:uh O;kl) Xyklowy] ,ywfeuk (800&200 esl) csathu esa feykdj ,d NksVkØksesVksxzkiQh dkWye rS;kj djsa rkfd lkiQ gqvk ,ywfeuk V~;wc dk2@3 Hkkx Hkj nsaA dkWye dks csathu ds lkFk Hkj nsa vkSj jcM+vkÅVysV V~;wc ;k DySi dk iz;ksx djrs gq, mldh fudyusdh xfr yxHkx 30 cwnsa izfr feuV ij fuf'pr djsa tc rd fdcsathu dh ijr ,ywfeuk dh lrg ls 2&3 feeh- Åij gksA

fVIi.kh µ ;g lykg nh tkrh gS fd ,ywfeuk ds Åijh Hkkx esa Xyklowy dk Iyx yxk;k tk, rkfd tc ?kksy feyk;k tk, rks mldhrhozrk dks jksdk tk lds vkSj dkWye ds Åijh Hkkx ij ?kksy dk lap;fd;k tk lds] tks ØksesVksxzkfiQd dkWye ij ekewyh ncko cuk,xkAlks[kus dh izfØ;k dks è;kuiwoZd fu;af=kr fd;k tkuk pkfg, rkfd,ywfeuk esa pSufyax dks jksdk tk ldsA csathu ;k vU; ?kksy dks ,Y;wVdjus ls iw.kZr% cpk tkuk pkfg,A ;g vko';d gS fd ,ywfeuk dhlrg dks nzoh; ijr ls

4

vkbZ,l@IS 2557 : 1994

acidic to pH test paper (pH 1-2). Dissolve the redprecipitate of norbixin by mixing the solution with50 ml of benzene. Discard the water layer and washthe benzene phase with water until it no longer givesan acid reaction. Remove any undissolved norbixin bycentrifugation or filtration and dry the solution overanhydrous sodium sulphate. Transfer 3-5 ml of the drysolution to the top of an alumina column prepared asdescribed above. Elute the column with benzene, three10-ml volumes of dry chloroform, followed by 5 mlof Carr-Price reagent to the top of the column. Theorange-red band of norbixin immediately becomesblue-green.

4.4.3 Thin Layer Chromatography

4.4.3.1 Annatto extract in oil

Use silica gel with 12 percent calcium sulphate asbinding agent, and a mixure of acetic acid, chloroformand acetone (1 : 50 : 50, by volume) as developingsolvent. The sample in oil produces at least 3 or 4 redor yellow spots. Two red spots are clearly more intensethan the other spots. Dissolve the two spots in benzeneand identify the spot that is bixin by the method givenin 4.4.2.2.

4.4.3.2 Water soluble annatto

Prepare a chromatogram as indicated under 4.4.3.1.Three or four spots are obtained, of which 2 spots havean orange colour and the others an orange-yellowcolour. Dissolve the orange spots in benzene andidentify one of the spots that is norbixin by the methodgiven in 4.4.2.3.

5 PURITY TESTS

5.1 The material shall conform to the requirementsgiven in Table 1.

LFkkukarfjr djsa vkSj i;kZIr 2 ,u lYÝ;wfjd ,flM feyk,aA50 feyh- csathu esa ukWjfcfDlu dh yky izsflfiVsV feyk,aAikuh dh ijr dks gVk nsa vkSj csathu iQst dks rc rd ?kksrs jgsatc rd og ,flM dh izfrfØ;k ugha nsrkA ukWjfcfDlu osQ uèkqys Hkkx dks Nyuh }kjk Nkudj fudky nsa vkSj ?kksy dks,ugkbMªksl lksfM;e lYiQsV ij lq[kk,aA tSlk fd Åij crk;kx;k gS 3&5 feyh- lw[ks ?kksy dks rS;kj fd, x, ,yqehukdkWye ds Åijh Hkkx esa LFkkukarfjr djsaA csathu ds lkFk dkWyedks fgyk,aA Mªk; DyksjksiQkeZ dh 10 feyh- ek=kk dks 3 ckj HkjsavkSj fiQj dkWye ds Åijh Hkkx esa 5 feyh- dkj&izkbl fjtsaVfeyk,aA ukWjfcfDlu dk dsljh&yky jax rRdky uhyk&gjk cutkrk gSA

4-4-34-4-34-4-34-4-34-4-3 ØksesVksxzkiQh dh iryh ijr

4-4-3-14-4-3-14-4-3-14-4-3-14-4-3-1 rsy esa vUukVks vdZ

lekèkku dks Li"V djus osQ fy, flyhlk tSy esa 12 izfr'krdSfY'k;e lYiQsV] ,flfVd ,flM] DyksjksiQkeZ rFkk ,dVksu(1 % 50 % 50 ek=kk) ds ?kksy dks feyk,aA rsy esa uewuk 3 ;k4 yky vFkok ihys èkCcs NksM+rk gSA nksuksa yky èkCcs Li"Vr%vU; èkCcksa ls T;knk rhoz fn[kkbZ iM+rs gSaaA nks èkCcksa dks csathuesa ?kksy nsa vksj fcfDlu èkCcs dh igpku 4-4-2-24-4-2-24-4-2-24-4-2-24-4-2-2 esa nh xbZi¼fr ds vuqlkj djsaA

4-4-3-24-4-3-24-4-3-24-4-3-24-4-3-2 ikuh esa ?kqyu'khy vUukVks

4-4-3-1 4-4-3-1 4-4-3-1 4-4-3-1 4-4-3-1 ds varxZr n'kkZ, x, ØksesVksxzke dks rS;kj djsaA 3 ;k4 èkCcs fn[kkbZ nsaxs ftlesa ls nks èkCcksa dk jax dslfj;k vkSjvU; èkCcksa dk jax dslfj;k&ihyk gksxkA csathu esa dslfj;kèkCcs dks ?kksy nsa ,oa 4-4-2-34-4-2-34-4-2-34-4-2-34-4-2-3 esa nh xbZ i¼fr ds vuqlkj mlèkCcs dh igpku djsa tks ukWjfcfDlu gksA

5 'kq¼rk ijh{k.k5 'kq¼rk ijh{k.k5 'kq¼rk ijh{k.k5 'kq¼rk ijh{k.k5 'kq¼rk ijh{k.k

5-15-15-15-15-1 lkexzh lkj.kh 1 esa nh xbZ vis{kkvksa ds vuq:i gksxhA

lkj.kh 1 vUukVks ds fy, vis{kk,¡lkj.kh 1 vUukVks ds fy, vis{kk,¡lkj.kh 1 vUukVks ds fy, vis{kk,¡lkj.kh 1 vUukVks ds fy, vis{kk,¡lkj.kh 1 vUukVks ds fy, vis{kk,¡([kaM 5-1)

y{k.k vis{kk,¡ ijh{k.k dh i¼fr_ lanHkZ ns[ks a Øe

la[;k bl ekud dk

vuqcaèk vkbZ,l 1699 %

1994 dk [kaM ns[ksa (1) (2) (3) (4) (5)

i) dSjVksuksbMl 0-24 [k — d) rsy esa vUukVks vdZ] fcfDlu ds :i esa vfHkO;Dr ek=kk] izfr'kr] U;wure [k) ikuh }kjk ?kqyu'khy vUukVks] ukWjfcfDlu ds :i esa vfHkO;Dr ek=kk] izfr'kr] U;wure 0-24 [k —

ii) vkjlsfud] fexzk-@fdxzk-] vfèkdre 3 — 15 iii) yhM] fexzk-@fdxzk-] vfèkdre 10 — 15 iv) dkWij fexzk-@fdxzk-] vfèkdre 30 — 15 v) Hkkjh èkkrq] fexzk-@fdxzk-] vfèkdre 40 — 16

5

vkbZ,l@IS 2557 : 1994

Table 1 Requirements for Annatto(Clause 5.1)

Characteristic Requirement Method of Test, Ref to Sl No.

Annex to This Standard

Ref to Clause of IS 1699 : 1994

(1) (2) (3) (4) (5)

i) Carotenoids a) Annatto extract in oil, expressed as bixin, percent by mass, Min 0.24 B — b) Water-soluble annatto, expressed as norbixin, percent by mass, Min 0.24 B — ii) Arsenic, mg/kg, Max 3 — 15 iii) Lead, mg/kg, Max 10 — 15 iv) Copper, mg/kg, Max 30 — 15 v) Heavy metals, mg/kg, Max 40 — 16

6 iSfdax] fpÉkadu vkSj HkaMkj.k6 iSfdax] fpÉkadu vkSj HkaMkj.k6 iSfdax] fpÉkadu vkSj HkaMkj.k6 iSfdax] fpÉkadu vkSj HkaMkj.k6 iSfdax] fpÉkadu vkSj HkaMkj.k

6-1 iSfdax6-1 iSfdax6-1 iSfdax6-1 iSfdax6-1 iSfdax

lkexzh dks mi;qDr fMCcksa esa tgka rd laHko gks de ls degok dh xqatkbZ'k gks] rkfd èkkrq ;k vU; v'kq¼rkvksa esa lkexzhdks nwf"kr gksus ls jksdk tk lds vkSj laHkor% izdk'k ls cpktkuk pkfg,A

6-2 fpÉk adu6-2 fpÉk adu6-2 fpÉk adu6-2 fpÉk adu6-2 fpÉk adu

6-2-1 6-2-1 6-2-1 6-2-1 6-2-1 izR;sd fMCcs ij Li"V o vfeV :i ls fuEufyf[krlwpuk,a nh tkuh pkfg,a%

d) lkexzh dk uke vkSj izdkj_

[k) mRiknu dk Ïksr_

x) U;wure fuoy nzO; ek=kk_

?k) cSp ;k dksM uacj_

Ä) mRiknu dh frfFk_

p) fcfDlu rFkk ukWjfcfDlu ek=kk dh ?kks"k.kk_

N) bl vk'k; dh fVIi.kh dh lkexzh BaMs LFkku ijj[kh tk,] vkSj dkap ds crZu dh fLFkfr esa izdk'kls nwj j[kk tk,_ vkSj

t) vU; dksbZ fooj.k tSls rkSy ,oa eki ekudksa (fMCckcanoLrq,a) fu;e] 1977@[kk| vifeJ.k fuokj.k fu;e]1955 ds varxZr mfYyf[krA

6-2-2 6-2-2 6-2-2 6-2-2 6-2-2 HkkekC;wjks izek.ku eqgjkadu

mRikn ij ekud eqgj Hkh yxkbZ tk ldrh gSA

6-2-2-16-2-2-16-2-2-16-2-2-16-2-2-1 ekud eqgj dk mi;ksx Hkkjrh; ekud C;wjks vfèkfu;e]1986 vkSj mlosQ vUrxZr cuk, x, fu;eksa vkSj fofu;eksa osQizkoèkkuksa osQ vUrxZr fu;af=kr fd;k tkrk gSA fofuekZrkvksavFkok mRikndksa dks ftu 'krks± osQ vèkhu ekud eqgj osQmi;ksx osQ fy, ykblsal fey ldrk gS] mudk fooj.kHkkjrh; ekud C;wjks ls izkIr fd;k tk ldrk gSA

6 PACKING , MARKING AND STORAGE

6.1 Packing

The material shall be filled in suitable containers withas little air space as possible so as to precludecontamination of the contents with metals or otherimpurities, and should preferably exclude light.

6.2 Marking

6.2.1 Each container shall be marked legibly andindelibly to give the following information:

a) Name and type of the material;

b) Source of manufacture;

c) Minimum net fluid contents;

d) Batch or code number;

e) Date of manufacture;

f) Declaration of bixin and norbixin content;

g) A statement to the effect that the materialshould be kept in a cool place, and in the caseof glass containers, away from light; and

h) Any other details as specified under theStandards of Weights and Measures(Packaged Commodities) Rules, 1977/Prevention of Food Adulteration Rules, 1955.

6.2.2 BIS Certification Marking

The product may also be marked with the StandardMark.

6.2.2.1 The use of the Standard Mark is governed bythe provisions of the Bureau of the Indian StandardsAct, 1986 and the Rules and Regulations madethereunder. The details of conditions under which thelicence for the use of Standard Mark may be grantedto manufacturers or producers may be obtained fromthe Bureau of Indian Standards.

6

vkbZ,l@IS 2557 : 1994

6-3 Hk aMkj.k6-3 Hk aMkj.k6-3 Hk aMkj.k6-3 Hk aMkj.k6-3 Hk aMkj.k

lkexzh dks BaMs LFkku ij j[kk tk,xkA

7 uewus7 uewus7 uewus7 uewus7 uewus

7-17-17-17-17-1 ijh{k.k ds fy, lkexzh ds izfrfufèk uewus vkbZ,l 1699%1994 ds 44444 esa mfYyf[kr i¼fr ds vuqlkj izkIr fd, tk,axsA

8 ijh{k.k8 ijh{k.k8 ijh{k.k8 ijh{k.k8 ijh{k.k

8-1 8-1 8-1 8-1 8-1 ijh{k.k lkj.kh 1 ds dkWye 4 ,oa 5 esa fd, x, mYys[kds vuqlkj fd, tk,axsA

8-2 vfHkdeZdks a dh xq.koÙkk8-2 vfHkdeZdks a dh xq.koÙkk8-2 vfHkdeZdks a dh xq.koÙkk8-2 vfHkdeZdks a dh xq.koÙkk8-2 vfHkdeZdks a dh xq.koÙkk

;fn fofufnZ"V u fd;k x;k gks] vU;Fkk ijh{k.k esa 'kq¼ jlk;uvkSj fMfLVyM okWVj dk mi;ksx fd;k tk,xk (vkbZ,l 1070%1992 ns[ksa)A

fVIi.kh — ^'kq¼ jlk;uksa* ls rkRi;Z mu jlk;uksa ls gS] ftuesa osvinzO; ugha gSa tks fo'kys"k.k ds ifj.kkeksa dks izHkkfor djrs gSaA

6.3 Storage

The material shall be stored in a cool place.

7 SAMPLING

7.1 Representative samples of the material for the testsshall be drawn according to the method prescribed in4 of IS 1699 : 1994.

8 TESTS

8.1 Tests shall be carried out as prescribed in col 4and 5 of Table 1.

8.2 Quality of Reagents

Unless specified otherwise, pure chemicals anddistilled water (see IS 1070 : 1992) shall be employedin tests.

NOTE — ‘Pure chemicals’ shall mean chemicals that do notcontain impuries which affect the experimental results.

vkbZ,l la- 'kh"kZd

75 % 1973 vylh dk rsy] dPpk ,oa 'kq¼ (nwljkiqujh{k.k)

231 % 1957 ,fey ,flVsV250 % 1964 iksVsf'k;e ckbZØksesV] rduhd vkSj

fo'ys"k.kkRed fjtsaV (iqujhf{kr)266 % 1993 lYÝ;wfjd ,flM (rhljk iqujh{k.k)296 % 1986 lksfM;e dkcksZusV] ,ugkbMªl (rhljk iqujh{k.k)376 % 1986 lksfM;e gkbMªksDlkbM] ,ukfyfVdy jhtsaV

(rhljk iqujh{k.k)542 % 1968 ukfj;y rsy (nwljk iqujh{k.k)544 % 1968 ewaxiQyh dk rsy (nwljk iqujh{k.k)546 % 1975 ljlksa dk rsy (nwljk iqujh{k.k)547 % 1968 lhle dk rsy (nwljk iqujh{k.k)1070 % 1992 fjtsaV xzsM okWVj (rhljk iqujh{k.k)1699 % 1994 [kk| jaxksa ds uewuksa vkSj ijh{k.k ds fy,

izfØ;k (nwljk iqujh{k.k)

IS No. Title

75 : 1973 Linseed oil, raw and refined (secondrevision)

231 : 1957 Amyl acetate250 : 1964 Potassium bichromate, technical and

analytical reagent (revised)266 : 1993 Sulphuric acid (third revision)296 : 1986 Sodium carbonate, anhydrous (third

revision)376 : 1986 Sodium hydroxide, analytical reagent

(third revision)542 : 1968 Coconut oil (second revision)544 : 1968 Groundnut oil (second revision)546 : 1975 Mustard oil (second revision)547 : 1968 Seasame oil (second revision)1070 : 1992 Reagent grade water (third revision)1699 : 1994 Methods of sampling and test for

food colours (second revision)

vuqcaèk dvuqcaèk dvuqcaèk dvuqcaèk dvuqcaèk d([kaM 2)

lanfHkZr Hkkjrh; ekudka s a dh lwphlanfHkZr Hkkjrh; ekudka s a dh lwphlanfHkZr Hkkjrh; ekudka s a dh lwphlanfHkZr Hkkjrh; ekudka s a dh lwphlanfHkZr Hkkjrh; ekudka s a dh lwph

ANNEX A(Clause 2)

LIST OF REFERRED INDIAN STANDARDS

7

vkbZ,l@IS 2557 : 1994

vkbZ,l la- 'kh"kZd

2491 % 1972 [kk| lalkèku bdkb;ksa dh LoPNrk fLFkfrds fy, dksM (igyk iqujh{k.k)

3490 % 1965 ukbZaxjlhM rsy3491 % 1965 dqlqe cht rsy

IS No. Title

2491 : 1972 Code for hygienic conditions for foodprocessing units (first revision)

3490 : 1964 Nigerseed oil3491 : 1965 Safflower oil

[k[k[k[k[k&&&&&1 rsy esa vUukVks vdZ1 rsy esa vUukVks vdZ1 rsy esa vUukVks vdZ1 rsy esa vUukVks vdZ1 rsy esa vUukVks vdZ

[k[k[k[k[k&1-1 1-1 1-1 1-1 1-1 uewus dks 0-1 ls 1 xzke rd ek=kk ds 100 feyh-okWY;we ds ÝykWLd esa Bhd ls uki dj LFkkukarfjr djsaADyksjksiQkeZ esa ?kksysa] DyksjksiQkeZ okys okWY;we esa feykdj feDldjsaA bl ?kksy dk ,d feyh- Hkkx nwljs 100 feyh- okWY;weokys ÝykLd esa LFkkukarfjr djsa rFkk 100 feyh- ?kksy ysaA

470 ,u,e osoyssaXFk ij bl ?kksy osQ ,CtkosZal , dks ekisaA

dqy izfr'krdkWjsVsukbZM~l × ×

=, 1000 000

2 -826 uewuk legw (fexkz -) 100(fcfDlu ds:i esa vfHkO;Dr)

[k[k[k[k[k&&&&&2 ikuh es a ?k qyu'khy vUukVks2 ikuh es a ?k qyu'khy vUukVks2 ikuh es a ?k qyu'khy vUukVks2 ikuh es a ?k qyu'khy vUukVks2 ikuh es a ?k qyu'khy vUukVks

[k[k[k[k[k&2-12-12-12-12-1 uewus dk 0-1 ls 1 xzke dk lgh rksy djsa] 100feyh- esa 0-01 ,u lksfM;e gkbMªksDlkbM feyk,a vkSj mlsiw.kZr% fgyk,aA bl ?kksy dk 1 feyh- Hkkx vyx iQuy esaLFkkukarfjr djsa vkSj mlesa 10 feyh- lksfM;e DykjksbM dk?kksy (10 esa 1) feyk,a vkSj 50 feyh- ikuh feyk,a vkSjmlesa 2 feyh- ?kqyk gqvk lYÝ;wfjd ,flM feyk,aA csUthu dsizR;sd 10 feyh- ds ?kksy dk vdZ fudkyrs jgsa tc rd fdcsathu vdZ jaxhu ugha gks tkrkA csathu vdZ dks feyk ysa]izR;sd 5 feyh- ikuh ls mls 3 ckj èkks,a vkSj ikuh dh ijrvyx gksus nsaA csathu vdZ dks ftlesa 2 feyh- csathu dh 3ikuh okyh ijrsa ikuh okys nwljs vU; iQuy esa LFkkukarfjrdjsaA bls csathu vdZ esa feyk nssaA bl csathu vdZ esa iSVªksfy;ecsathu feDl dh leku ek=kk feyk,a vkSj 0-01 ,u lksfM;egkbMªksDlkbM izR;sd 5 feyh- vdZ feykrs jgsa tc rd fdlksfM;e gkbMªksDlkbM ?kksy jaxhu ugha gks tkrkA bu ?kksyksa dksbdV~Bk djsa vkSj 100 feyh- esa 0-01 lksfM;e gkbMªksDlkbMfeyk nsaA 453 ,u,e osoysaXFk ij ,CtkosZal , dk eki ysaA

B-1 ANNATTO EXTRACT IN OIL

B-1.1 Transfer 0.1 to 1 g of the sample, accuratelyweighed, into a 100-ml volumetric flask. Dissolve inchloroform, dilute to volume with chloroform, and mix.Transfer a 1 ml portion of the solution into another100-ml volumetric flask, and dilute to 100 ml.

Measure the absorbance A of this solution at thewavelength of 470 nm.

Percent totalcarotenoid

1 000 000×

2.826 Sample mass (mg) × 100�

A

(expressed asbixin)

B-2 WATER-SOLUBLE ANNATTO

B-2.1 Weigh accurately 0.1 to 1 g of the sample, add0.01 N sodium hydroxide to 100 ml, and shakethoroughly. Transfer 1 ml portion of the solution into aseparating funnel, add 10 ml of sodium chloridesolution (1 in 10) and water to 50 ml, and add 2 ml ofdiluted sulphuric acid. Continue to extract the solutionwith each 10 ml of benzene until the benzene extract isnot coloured. Combine benzene extract, wash threetimes with each 5 ml of water, and allow to stand toseparate the water layer. Transfer the benzene extractinto another separating funnel containing the water layerthree times with 2 ml of benzene, and combine thewashings to the benzene extract. To the benzene extract,add an equal volume of petroleum benzene mix, andcontinue to extract with each 5 ml of 0.01 N sodiumhydroxide until sodium hydroxide solution is notcoloured. Combine extracts, and add 0.01 sodiumhydroxide to 100 ml. Measure the absorbance A at thewavelength of 453 nm.

vuqcaèk [kvuqcaèk [kvuqcaèk [kvuqcaèk [kvuqcaèk [k¹lkj.kh 1] Øe la[;k (i)º

dkWjVsukbZMl dk fuèkkZj.kdkWjVsukbZMl dk fuèkkZj.kdkWjVsukbZMl dk fuèkkZj.kdkWjVsukbZMl dk fuèkkZj.kdkWjVsukbZMl dk fuèkkZj.k

ANNEX B[Table 1, Sl No. (i)]

DETERMINATION OF CAROTENOIDS

8

vkbZ,l@IS 2557 : 1994

fuoZpu esa fookn dh fLFkfr esa bl ekud dk vaxzsT+kh ikB gh ekU; gksxkAIn case of dispute in interpretation, English version of this standard shall be authentic.

dqy izfr'krdkWjsVsukbZM~l

(ukWjfcfDlu ds:i esa vfHkO;Dr)

Percent totalcarotenoid

1 000 000×

3.473 Sample mass (mg) × 100�

A

(expressed asnorbixin)

××

= , 1000 0003- 473 uewuk legw (fexkz -) 100

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Review of Indian Standards

Amendments are issued to standards as the need arises on the basis of comments. Standards are also reviewedperiodically; a standard along with amendments is reaffirmed when such review indicates that no changes areneeded; if the review indicates that changes are needed, it is taken up for revision. Users of Indian Standardsshould ascertain that they are in possession of the latest amendments or edition by referring to the latest issue of‘BIS Catalogue’ and ‘Standards : Monthly Additions’.

This Indian Standard has been developed from Doc No.: FAD 8 (256).

Amendments Issued Since Publication

Amend No. Date of Issue Text Affected

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Published by BIS, New Delhi