Disclosure to Promote the Right To Information

Whereas the Parliament of India has set out to provide a practical regime of right to information for citizens to secure access to information under the control of public authorities, in order to promote transparency and accountability in the working of every public authority, and whereas the attached publication of the Bureau of Indian Standards is of particular interest to the public, particularly disadvantaged communities and those engaged in the pursuit of education and knowledge, the attached public safety standard is made available to promote the timely dissemination of this information in an accurate manner to the public.

इंटरनेट मानक

“!ान $ एक न' भारत का +नम-ण”Satyanarayan Gangaram Pitroda

“Invent a New India Using Knowledge”

“प0रा1 को छोड न' 5 तरफ”Jawaharlal Nehru

“Step Out From the Old to the New”

“जान1 का अ+धकार, जी1 का अ+धकार”Mazdoor Kisan Shakti Sangathan

“The Right to Information, The Right to Live”

“!ान एक ऐसा खजाना > जो कभी च0राया नहB जा सकता है”Bhartṛhari—Nītiśatakam

“Knowledge is such a treasure which cannot be stolen”

“Invent a New India Using Knowledge”

है”ह”ह

IS 7375 (1979): Salseed Fat [FAD 13: Oils and Oilseeds]

IS : 7375 • 1979

Indian StandardSPEClf-ICATJON FOR SALSEED FAT

( First Revision)

riO~&!::JI

INDIAN STAN1)ARDS INSTITtJTION~ft\NAI~ Hl-it\V.'\N, ~~ HAlfAl)lYR ~'rlAB ZAFAH. ~l;\H.t~

KI:. Vf IJr,Ll:l1 i }11\Ji'2

IS I 7375 • 1979

Indian StandardSPECIFICATION FOR SALSEED FAT

( First Revision)Oils and Oilseeds Sectional Committee, CAFDC 5

CluJi,m.aPao)' M. M. CHAKRABARTY

R,"'I,tdi",Oil Technologilts' Association or India, Kanpur

Central Organization for Oil IndUJtry & Trade,Bombay

M,,,.6,,,PRO)' A. C. GUPTA ( A/I""at, to

Prof M. M. Chakrabarty)SaBI S. N. AGARWAL

SURI H. P. GUrJ'A ( AII,rnal, )AGRICULTUBAL MARKF::TINO Directorate or Marketing & Inspection (Ministry of

ADVISER TO TS. GOVBRNIf£N'r Agriculture), NagpurOJ'INDIA

SIIRI T. V. MATHEW ( All"nat, )Snut D. A. ASRTAPUTltE Ministry or Defence ( DGI )

SSRI p. N. AOAI\WAL ( Alt",",t, )Da B. P. BALIGA Tata Oil Mills Co Ltd, Bombay

DR N. L. MUUTY ( Alt"nal, )SaRI M . .1\. BUA'I'T Swastik Household Be Industrial Products Ltd,

BombaySHRI C. R. KRI8RNAMURTHY

( Alt"nal, )I>K N. V Bn,INOl Hindustan Lever Ltd, Bombay

SHRI K BAI4AKRI8HNAN (Alter,.llI,)SURJ D. S. CHADIIA Central Oommirtee for Food Standards (Directorate

Genera] of Health S~rvices ). New DelhiSMT DEDI MUKIIRRJEE ( .A11""11I' )

SDRI N. C. CHATTERJ~E National Test House, CalcuttaSlIRI B. V. KANTAX Godrej Soaps Ltd, Bombay

SHat M. S. THAXUI& ( Alt"nat, )SaRI R. D. KAWA'l'RA Directorate General of Technical Development,

New DelhiSHRI S. KRISHNAMUltTIlY Khadi at Village Industries Commi••ion, Bombay

SBRI V. LAK8HMIKANTHAN (AII,rnal, )On, G. LAK811MINAI&AYANA Regional Research Laboratory ( CSIR ), Ilyderabad

DR A. J. PANTULU ( Alte,.ale )

( ConlillUld Oft ,." 2 )

@ CO/J",;,lal 1980INDIAN STANDARDS INSTITUTION

This publication i. protected under the Indi." Co/J",i,hl .Ad ( XIV or 1957 ) aDdreproductioa in whole or in part by aDy means except with wriuea permillioD or tbepublisher shall be deemed to be aD iDfrinl('ment of copyright under the .aid Act.

II I 7375 • 1979

( Coralinu,tJ from pall I )

Directorate or Vanaspari, Vegetable Oi15 and Fats(Ministry of Commerce, Civil Supplies andCooperation ), New Delhi

Rt/Jr"enti",Indian Agricultural Research Institute, New DelhiItalab Pvt Ltd, Bombay

M,mb,r,DR S. K. l\loKllEnJBBSHRI J. P. PA'I'Jr.L

SHn T J. C. DRY ( Alt",.at~ )( Calcutta)

SHRI S. S. HONAVAli. ( Alt'rnat, )( Madras)

Dn N. S. RAJAOOPAL

On M. K. KUNf)tJ ( Altemat« )SHIH P, RAMI)AS Central Wareh()u~ingCorporation, New Delhi

SAUl A. S. SAlUPATRKUMAR ( Alurnat« )Dn Y. R. RAO Regional Research Laboratory (CSIR). Bhuba-

neshwarDR M. BAPU JI ( Altemat« )

SURI P. K. SJr.N Vanaspati Manufacturers' Association of India,Bombay

SHRI RAJ KAPILA ( Alternat» )Snnr HAIUSll SETHI Directorate of Oils..cds Development ( Ministry of

Agriculture ), HyderabadSURI G. V. SIRUB Solvent Extractor. Association of India, Bombay

SHR.I L. KIUSHAN KUMAR (Alt,rnal,)SHnI S. D. 'r.ltlltlTMAl,A RAO Oil Technological Research Institute, Anantapur

SHill n. A'l'GHYUTAltAMAYYA ( Alternate )DR G. M. SAXl~NA, Director General, lSI ( Ell-officio M,mber)

Director ( Chern )

Stcretary

SURt R. K. SJNOnDeputy Director ( Chern ), lSI

Oils and Fats Subcomrnittee, CAFDC 5 : 1

Conll,,,,r

DR G. LAKSIIMINAUAYANA Regional Research Laboratory ( CSIR ). Hyderabad

MembersSURI GOKUI.. CHAND J. AOARWAI, Bombay Oilseeds and Oils Exchange Ltd, Bombay

SHIU CUARANDAS V. MAIU-

W ALA ( Altemat« )AORICULTUltAJ.. MAllKEorING Directorate of Marketing &. Inspection ( Ministry

ADVIS"~n. TO 'l'HI·~ GOVERNMKNT of Agriculture- ), NagpurOJ'INDIA

SHl~I rr. V. MA'l'UI£W ( Altemat» )SUBI D. A. ASH'l'APl;TRE Ministry of Defence ( DGI )

SHRI P. K. MAJUMDAR ( Alternat« )SSRI D. S. CHAUHA Central Committee for Food Standards ( Directorate

General of Health Service, ), New DelhiSM'r 0 J~BI M 1: K 11E ItJEE ( Alu,not, )

( Continued Oft /Jag, 14 )

2

IS I 7375 • 1979

( Conti"u,d from /JaIl 2 )

M,mIJ", R,p"s,ttti",DBA. C. CUATBAPATI Vanaspati Manufacturers' Association or India,

. BombaySSBI C. NARAYANA.WAIlY ( Alt""at,)

SDRI R. D. KAWATRA Directorate General or Technical Development,New Delhi

Indian Council of Agricultural Research, New DelhiKhadi & Village Industries Commission, Bombay

( Alurnat« )Indian Paint Association, Calcutta

DR B. M. LALSHRI V. LAK8HMJXANTSAN

8111\1). N. BHATTACHARYASHBI R. K. MABPHA1'IA

SHUl M. N. RAO ( Alt,rnat, )Du N. L. MURTY Tata Oil Mills Co Ltd, Bombay

DR A. D. SHITOLB ( Alt,rnatt )DB S. M. PATEL Oil Technologist.' Association of India, Kanpur

PaOF V. V. R. SUBRAMANYAM (AlI,rnat,)DB N. S. RAJAQOPAL Directorate of Vanaspat], Vegetable Oils and Fatl

( Minislry of Commerce, Civil Supplies and Co­operation), New Delhi

DB I. A. SIDDIQI ( AII,rna', )DB D. p. SEN

SBBJ HABI8R SETHISJlBI M. S. THAKURSUBI S. D. TDIRUIIALA RAO

Central Food Technological Research Institute( aSIR ). Mysore

Directorate of Oilseed. Development, HyderabadGodrej Soaps Ltd, BombayOil Technologieal Research Institute, Anantapur

14

AMENDMENT NO. 1 SEPTEMBER 1995TO

IS 7375: 1979 SPECIFICATION FOR SALSEED FAT( First Revision)

( Page 4, Foreword, clause 0.6 ) - Add the following clause 0.7 afterclause 0.6 and renumber the subsequent clause:

'0.7 A scheme for labelling environment friendly products to be known as EeDMark has been introduced at tbe instance of the Ministry of Environment andForests ( MEF). The ECO Mark shall be administered by the Bureau of IndianStandards ( BIS ) under the BIS Al-t, 1986 as per the Resolution No. 71 dated 20February 1991as published in the Gazette of the Government of India vide GSR85(E) dated 21 February 1991. For a product to be eligible for marking witb tbeECO Mark it shall also carry the Standard Mark of BIS for quality besidesmeeting additional optional environment friendly ( EF ) requirements, The EFrequirements for salseed fat are therefore being included through an amendment,

This amendment is based on tbe Gazette Notification No. 678 dated 30 August1994 for Labelling Edible Oils, Tea and Coffee as environment friendlyproducts, published by the' Ministry of Environment and Forests.'

(Page 5, clause4.5.1 ) - Add tbe following clauses after clause 4.5.1:

'4.6 ()ptional Requlrements for ECO Mark

4.6.1 General Requirements

4.6.1.1 The product shall conform to the requirements of quality prescribedunder clauses 4.1 to 4.5.

4.6.1.1 The manufacturers shall produce to BIS environmental consentclearance from the concerned Stale Pollution Control Board as per the norms laiddown under the Water ( Prevention and Control of Pollution) Act, 1974;Air ( Prevention and Control of Pollution ) ACl, 19Rl ; Waler ( Prevention andControl of Polllltion) Cess Act, 1977, respecuvely, along with theauthorization, if required under the Environment ( Protection) Act, 1986, whileapplying for ECO Mark.

4.6.2 Specific Requirements

Amend No.1 to IS 7375 : 1979

(Page 13, Fig. 2 ) - Add the following Appendix after Fig 2:

'APP..:NDIX It(Clouse 4.6.2.1 )

DE"fERMINATION 0)4" A.'I.Al'OXIN

B-1 REAGENTS

11-1.1 Acetone, 70 Percent - 700 ntl acetone in 30n ml distilled water.

li-t.Z Acetone, %0 Percent - 200 ml acetone in 800 ml distilled water.

8-1.3 Lead Acetate, 20 Percent - 200 g neutral acetate in distilled water and3 011 glacial acetic acid, diluted to one litre.

11-2 I)R()(~RDIJRE

0-2.1 Dissolve 30 g sample in 100 ml hexane.

11-2.2 Extract with 3 x 50 ml 70 percent acetone.

11·2.3 To the extract add 60 ml distilled water and 20 ml lead acetate.

11-2.4 Boil to reduce volume 10 150 ml. Cool to about 20°C.

8-2.5 Filter and wash with 20 percent acetone.

8-2.6 Extract filtrate and washings with 3 x 50 ml chloroform.

8-2.7 Passchloroform layer through anhydrous sodium sulphate.

11-2.8 Concentrate to 50 ml and spot on TLC plate.

11-] CAI.I(~lIIAl"I()N

Ail. or./1, V X S x 1 no{)

atoxin, In~ng =it X m

where

V =volume of extract in ml,

v = volume of extract giving minimum observable fluorescence in ul,

m =mass of sample in g, and

J =standard toxin giving minimum observable fluorescence in ....8.'

(FAD44 )RcprograpbyUnit, B1S. NewDelhi, India

3

AlDend No.1 to IS 7375: 1979

4.6.2.1 The product shall not contain ,flatoxin, more than 5 mglkg, when testedby the method prescribed in Appendix B.

4.6.1.1 The pesticide residues, if any, shaJl not exceed the tolerance limits asprescribed in the Prevention of Food Adulteration Act, 1954 and Rules madethereunder.

4.6.2.3 Only permitted antioxidants not exceeding the quantities specifiedagainst each 8S prescribed under the Prevention ofFood Adulteration Act, 1954and Rules made thereunder, shall be used, if required.

4.6.2.4 The product shall not contain any of the toxic metals in excess of tbequantities prescribed in Table 2..

l"AHLE 2 LIMITS ""OK TOXIC METALS

i) Lead, mg/kg. Max 5.0

il) Arsenic, lng/kg. Mf'X 05

iii) Cadmium, mglkg. Max 1.0

iv) M~rC'ury (total) 0.25mglkg. Max

• Methods of sampling and lest for food colours ( second revision ).

SL No. CHARAC11:.RJSTIC REOUlIU~f:.NT Mf11fODoFTt..~r, REFn)

IS of IS 16Q9: 1995·

do

do

do

( Page 6, clause S.1 ) - Add the following clause S.J.l after clause s.r.'5.1.1 For EeD Mark the product shall be packed in such packages which aremade from recyclable ( that is which can be re-processed to manufacture anyuseful product) or biodegradable materials.'

(Page 7, clause 6.1 ) - Add the following clause '.3 after clause 6.1:

"6.3 For ECO Mark the containers shall be marked with the followinginformaIi011:

a) List of identified critical ingredients in descending order of quantity,percent by mass, which sball include 'made from salseed fat';

b) The brief criteria for which the product has been labelled for ECO Mark;and

c) Sbelf life of tbe product. t t

2

AMENDMENT NO.2 MARCH 2002TO

IS 7375: 1979 SPECIFICATION FOR SALSEED FAT( First R~visioll )

(Anlendment No.1, page 2, clause 4.6.2.1 ) - Substitute '5 J,lglkg' for'5 mglkg t.

(FAD 44 )

Rcprography Unit. BIS. New Dellu, India

IS : 1375 • 1979

Indian StandardSPECIFICATION FOR SALSEED FAT

(First Revision)

e, FOR E W 0 R D

0.1 This Indian Standard ( First Revision) was adopted by the IndianStandards Iustitution on 26 Sr ptcmber 1979, after the draft finalized bythe Oils and Oilseeds Sectional Committee had been approved by theChemical Division Council and the Agricultural and Food ProductsDivision Council.

0.2 Salseed fat is obtained frorn the Sf eel kernels of sal tree (Shorearobusta Gaertn, f. ) widely known as sa I in north... r n India, sal tree is oneof the outstanding forest trees of Uttar Pradesh, Madhya PIadesh, Bihar,O. issa, West Bt ngal and Assam and its other common Indian namesarc sal ( Bengal ), Belsal, Borsal, Borsar (Galo HIlls, Assam ), Sarai (M P.)Salwa, Sektoa, Sorin..l!hi, Saragi, Rangal ( Orissa ), Saluia, T'amba, Sariakamu( T clugu ). In Indian liter ature it is also known as Asuiakarna. Thetotal potential of salsevd fat is very large in the country.

0.3 This standard was first puhlished in 1974. Salseed kernels containup to 18°5 percent «f a glt~lli~h hard fat. The fat derived fromseeds of Shorea robusta (;clrrtn. f. is berng use-d as a CO( oa butter substitute,The salseed fat has also bven tr.idit iona lly used by JO( a] population fore-dible use. It has hf'l'll cxpor te d to many couuu ies in rer cnt past for useas a cocoa butter su bstnutr-, The stud it"~ under taken so f~\r 011 itssuitability fot edible uses have lx en found grtlet ally sat J:-.fdctOl y. Theconr-er ne.l techlJical comm itn-e, therefor c, decided to I( vise this standardincorporating refined sa lsce-d fa t for edible US( So

0.3.1 The usr of refilled sa lseerl fat in the manufacture of bakerv andconfect ionary p' odur ts is being pel rnittr-d uncle r the P, t vr-nt i.m l'( FoodAdulteration ( P~"'A ) Act by GoveJ nrnr nt of Inrli ..l. IIc1\\('vCJ, its use forother eelible pur poses, such a~ in the rna nufactur (' of vanaspat i is underact ive consider arion of the Ministi y of Hr-alth, Government of Ir.dia.

0.4 The oleic acid content of salsec d f~lt a nd cocoa butter 31 e almostidentical, at ellound 40 pt'l cent. The satui c.u{'d acids in cocoa butter at eopalmitic and stear ic ill roughly (qual propol rions, thus giving rise to ahigh propor t ion of mixed gl)c{otidt s conra iniug all these tlut r ;ICJds. Insalsced fat, the saturated acid is almost e-ntir ely stearic It admg mostly to

3

18 I 7375· .979

stearic oleic glycerides. Disaturated glycerides predominate in both fatsgiving them a certain similarity in properties.

0.5 This standard contains clause 5.1 which calls for agreement betweenthe purchaser and the supplier.

0.6 In the preparation of this revision, substantial assistance has beendrawn from data supplied by Directorate of Oilseeds Development,Hyderahad; Hindustan Lever Ltd, Bombay; Khadi & Village Indu-n iesCommission, Bombav; Oil Technological Research Institute, Anarn .pur(AP) and Rcgion.vl Research Laboratories, Bhubaneswar and Hyder ..badand the assistance so derived is thankfully acknowledged.

0.7 For the purpose of deciding whether a particular requirement ofthis standard is complied with, the 611,\1 value, observed or calculated,expressing the result of a test or analysis, shall be rounded off inaccordance with IS : 2-1960*. The number of significant places retainedin the rounded off value should be the same as that of the specifiedvalue in this standard. -

1. SCOPE

1.1 TIlls standard prescribes requirements and methods of sampling andtest for s.ilseed fat.

2. TERMINOLOGY

2.1 For the purpose of this standard, the definition ~iven under 2 ufIS : 548 ( Part I )-1964t and IS : 548 ( Part III )-1976:, and the follow­ing shall apply.

2.1.1 Raw Salseed Fat - Salseed fat obtained by the process of solventextraction.

2.1.2 Refined Salseed Fat - Salseed flit obtained by the process of solventextraction which has been neutralized with alk.rli , bleached withbleaching earth or activated carbon or both, and deodorized with steam,no other chemical agents being used. Alternatively de-acidificntion,bleaching and deodorization may be done by physical means.

• Rules for rounding off numerical values ( "lIis,d ).tMerhods of sampling and test for oils and fats: Part I Methods of sampling. physical

and chemical tests ( "vi"d ).tMethod. of sampling and tell for oils and fats: Part III Analysis by gas liquid

chrornatography.

4

IS I 7375 • 1979

3. GRADES

3.1 The material shall be of the following three grades:

a) Refined grade,

b) Raw grade 1, and

c) Raw grade 2.

3.1.1 The material of the refined grade is suitable for edible pur poses( SI' also 0.3.1 ).

3.1.2 The material of raw grade 1 is suitable for making vanaspati,refined fat and for use in bakery and confectionary products only, andnot for direct consumption.

3.1.3 The material of the raw grade 2 is suitable for industrial uses.

4. REQUIREMENTS

4.1 The material shall be obtained from clean and sound seed kr-rnels ofsal n (~C, Shorea robusta Gaertn. f. (N.O. Dipterocarpaccac ) by the processof solvent extraction.

4.1.1 Solvent-extracted oil shall b 10 obtained from the seed kernelsusing solvent hexane conforming to IS : 3470·1966·.

4.2 The material of refined grade shall be clear on Inciting and free fromadulterants, rancidity sediment, suspended and other foreign matter,separated water, and added colouring or flavouring substances and shallhave acceptable taste and odour. It may contain permitted antioxidantsin specified quantities as prescribed under the Prevention of Food Adul­teration ( PFA ) Rules of the Government of India,

4.3 The clar ity of the material shall be judged by the absence of anyturbidity after keeping the melted filtered sample at 40°0 for 24 hours.

4.4 Admisture with Other Oils .- The material shall be free fromadrnixtur c with mineral oil and other oils when tested according to themethods prescribed in IS : 548 ( Part II )-1976t.

4.5 The material of refined grade and raw grade 1 shall also comply withthe J l quirt ments given in Table 1.

4.5.1 The material of raw grade 2 shall also comply with the require­me nts given in Table 1 for raw grade 1 except in cast' of arid valuew hich shall be maximum 20.

·Specification for hexane, food grade.t Methods of sampling and test for oils and fats: Part II Purity tests ( thi,d "visj.n J.

.1 I 7375 • 1979

TA.BLB 1 RBQUIREMBNT. POR SA.LSBED FAT( CI."s" 4.5 "tid 4.5.1 )

SLNo.

CBABAOTJ:BIITIO REQUIBBIIENT :rOB

,..---~----.

Refined RawGrade Grade 1

MB'1'BOD o:r TEsT,RBI' TO

"......---.j..----~Appen- OJ No.

dix

(1) (2) (3) (4) (5) (6)

i) Moisture and volatile matter. 0·1 1·0 5 &61percent by mass, M",~

ii) Refractive indext at 40°C 1-4500 to 1·4600 10 Iiii) Acid valuet, Mil" 0·5 12 7 lOrIS: 548iv) Iodine value ( Wijs ) +--35 to 45-..... 14 ( Part I)-

1964*v) Saponification value +--186 to 195-+ 15

vi) Titre, ·C -46 to 53 12

vii) Unsaponifiable matter, per- +---2'5----+ 8 Jcent by masl, MtI~

viii) Flash point, Pen sky-Martens 250 100 IS : 1448( closed), ·C, u« [P: 21 ]-19701

ix) 9. I0 epoxy and 9, 10 di- 3'0 Ahydroxy Itearic acids, per-cent by mall, MG~

*Methodl of sampling and test for oils and fats: Part I Methods of lamplinlphysical and chemical telta ( "",sId ).

tThis corresponds \0 8. R. reading of 36'7 to 51'0.*The corresponding figure in terms of free fatty acid. ( FF A ) when expressed as

oleic acid shan be maximum 0'25, 6 and 10 percent by mass for refined, raw grade 1 andraw grade 2 of la lseed rat respectively.

IMethods of telt for petroleum and its productsi P : 21 Flash point ( closed ) byPenlky.Martcnl apparatus <first "viJion).

s. PACKING

5.1 The material shall be packed in suitable well-closed containers asagreed to between the purchaser and the supplier. The refined grade orsalseed fat shall be packed in new tin containers.

6. MARKING

6.1 The containers shall be marked with the following information:

a) Name and grade of the material;b) Net mass of the material;

6

18 : 1:J75 -197.

c) A statement that permitted antioxidants have been used,if added;

d) Manufacturer's name and his recognized trade-mark, if any;

e) Batch number or lot number in code or otherwise; and

f) Month and year of manufacture.

6.1.1 The containers may also be marked with the lSI CertificationMark.

NOTE - The use of the lSI Certification Mark is governed by the provisions of theIndian Standards Institution ( Certification Marks) Act and the Rules and Regu­lations made thereunder. The lSI Mark on products covered by an Indian Standardconveys the assurance .hat they have been produced to comply with the requirementsof that standard under a well-defined system of inspection, testing and qualitycontrol which .. devised and supervised by IS I and operated by the producer. lSImarked product. are also continuously checked by lSI for conformity to that standardas a further safeguard. Detail. or conditions under wbich a licence for the use ofthe lSI Certification Mark may be granted to manufacturers or processors, may beobtained from the Indian Standarda IDltitution.

'.2 In addition, in case of raw grades of the material following infor­mation shall also be suitably marked, either printed on the label affixedto the container or lithographed or stencilled thereon with indelible inkin a type size of not less than 50 mm:

a) Raw grade 1 - 'NOT FOR DIRECT EDIBLE USE'

b) Raw grade 2 - 'FOR INDUSTRIAL NON-EDIBLE USESONLY'

7. SAMPLING

7.1 Representative samples of the material shall be drawn as prescribedunder 3 of IS : 548 ( Part I )-1964*.

7.2 All the characteristic. given in this specification shall be tested on thecomposite sample.

1.3 The lot shall be declared as conforming to the requirements of thisspecification, if all the test results on the composite sample satisfy therequirements given in 4.

*Methods or .amp\in~ and trlt lor oil. and fats: Part I Methods of Jamplinl.physical and chemical tests ( ".is,tl ).

7

IS I 7375 - 1979

8. TESTS

8.1 Tests shall be carried out as prescribed in IS : 548 ( Part I )-1964·,IS : 548 ( Part II )-1976t, IS: 1448 [ P : 21 ]-1970: and Appendix A.Reference to the relevant clauses of IS: 548 ( Part I )-1964*,IS : 1448 [ P : 21 ]-1970: and Appendix. A is given in col 5 and 6 ofTable 1 and that of IS : 548 ( Part II )-1976t in 4.4.

1.2 Q,1Iallty of Realents - Unless specified otherwise, pure chemicalsand distilled water ( se IS : I070-1977§ ) shall be used in tests.

NOTB -' Pure chemicals' shall mean chemicals that do not contain impuritie..which affect the results of analysis.

APPENDIX A[ Table 1, Item (ix) ]

ESTIMATION OF EPOXY AND DIHYDROXY STEARIC ACiDS

A-o.GENERALA-O.I Two methods have been prescribed. The method given in A-Ishall he used as a referee method whereas the method given in A·2 maybe used for routine analysis.

A-I. ESTIMATION BY GAS LIQ,UID CHROMATOGRAPHY(GLC)

A-I.O Outline of the Method - The method consists of enrichmentof triglyceridcs containing 1 mole of9,lO-epoxy stearic acid and 2 molesof predominantly stearic acid ( designated as P ) and other triglyceridecontaining 9,1O.dihydroxy stearic acid in place of epoxy stearic acid( designated as Q.) from a known mass of salseed fat by treatment withsilicic acid in hexane and desorbing these by a more polar solvent. Thedesorbate to which a known quantity of internal standard is added issubjected to transmethylation, and the methyl esters after silylationanalyzed by GLe.

• Methode of sampling and tesl for oils and fats: Part I Methods of aampliul,physical and chemical testa ( ',Dis,d ).

t~fethods of sampling and test for oil'] and fats: Part II Purity test ( thi,d ,'O;S;01l ).*Methods of test for petroleum and its products: P : 21 Flash point ( closed) by

Penlky-Marteol apparatus <fi,st"v;sion ).§Specification for water for general laboratory use ( "cond "lIis.ion ).

8

IS I 1375 - 197.

A-I.I Procedure

A-I.I.I Enrichment of 'P' and 'a: and T,ansm~thylation - Dissolve 200 mgof refined salseed fat in 2 ml of a-hexane and stir with 600 mg of silicicacid (activated at 110°0 for 1 hour before US(" ) for I·5 hours USil1 g amagnetic stirrer. Pipette out the supernatant hexane and wash the residuewith additional 2 rnl of hexane and discard the hexane, Add 2 ml of chloro­form methanol (3: 1 ) (vlv ) mixture to the residual silicic acid and stir for45 minutes. Filter the contents and wash the residue with additional2 X 2 ml chloroform-methanol mixture ( 3 : 1 ) (vllJ) to ensui e COIn pletedesorption of the adsorbed material, Add a known amount of about2 mg of n-heptadecanoic acid methyl ester to the combined chloroform­methanol filtrate contained in a 10 ml round bottom flask and evaporatesolvent to dryness under a stream of nitrogen. Dissolve the residue in1 ml of dichloromethane and retreat it with 2 ml of 2 M solution of sodiummethoxidc in methanol and keep at 50°C for 15 minutes with occasionalshaking. During this treatment connect the flask to a condenser andcalcium chloride guard tube, Dilute the contents with 2 ml saturat cdsodium chloride solution and extract with n-hcxane (3 X 2 ml ) in aseparating funnel. Wash the combined hexane extract with water toensure complete removal of alkali' ( by pI-I pape-r ), dry over anhydroussodium sulphate and evaporate to dryness under nitrogen.

A-I.l.2 Silylalion - Transfer the final residue tu a 5 1111 screw-cappedvial with the aid of ether and subsequently remove thr ether by evapor­ation under nitrogen. Dissolve this in 0·2 rnl of dry pyridine and treatwith 0·1 ml of hexamethyldisilazane and 0·05 ml of ch lorotrimethv lsilaneand keep at room temperature for 1 hour. Evaporate the reagents todryness on a water bath at about 50°C under nitrogen.

25 cm/h

60 ml/min

c) Temperature of inletoven and detector

d) Chart speede) Nitrogen flow

A-I.I.3 GLC Analysis - Dissolve thr- residue obtained in A-l.l.2 inhexane and inject into GLe colurnr rhr details of the instrument andconditions r see also IS : 548 ( Part 111 )-1976· ] shall be as follows:

a) D(,tt"ctor : Flame ionization detector ( FID )b) Colurnn 2·5 M X 4 turn glass; 1 percent

O\'-17t on 80 to 100 mesh GasChrorn Q

: 195°C

-Methods of sampling and test for oils and fats: Part III Analysis by gas' liquidchromatography.

tSupplied by Applied Science Laboratories, Pennsylvania, USA or equivalent"

9

II I 1375 • 1979

.\-1.2 CalealatloD

.\-1.2.1 A typical OLe scan i. shown in Fig. 1 in which the peaks dueto methyl esters of epoxystearic acid and silylated dihydroxy-stearic acidare marked. Calculate the amount of 'P' and 'Q; 81 follows ( s" Note ):

Peak area of epoxy Amount of'P', percent by mass _ acid methyl caster X internal X 904* x~

Peak area of the standard 312 massinternal standard in log ofsam-

ple inmg

Peak area of dihy- Amount ofen' t b droxy[ methyl ester X internal X 922: X 100)(." percen y mass - . d --Peak area of the standar 330 Mass of

Internal standard in mg samplein mg

NOTE - The GLe method or ' p. and' Q.t estimation is applicable to rtfinedrat or fats with free fatty acids ( FFA ) lesl than 2 percent. Ira eaee of hilh FFA fat,Deutralization should precede tr."selterificat;on. For this purpose spr.y a Imallquantity of fat with 3 N sodium hydroxide ( 10 percent excels) containing 10 percentsodium chloride at 50 to 60°0 under gentle stirring. Afler allowing the soap to­settle for a while transfer the material to a tube and then centrifuge. Wash theluperbatant oil free from soap and take far tranaesterification.

A-l.2.2 Epoxy and dihydroxy fatty acids,percent by mass

P+Qz:= -

3

A-2. ESTIMATION -r AND cQ' BY PREPARATIVE THIN LAYERCHROMATOGRAPHIC ( TLC ) METHOD

A-2.0 OutliDe or the Method - 'P' and 'Q: are separated on prepara­tive TLC plates and the bands scraped, extracted with solvent, evaporatedand weighed. This method is applicable to fats containing approximately3 percent each or more of 'P' and cQ'. Fats containing Jesser proporrionsof 'P' and ccr need enrichment prior to preparative TLC in order to g~t

reasonable amount of 'P' and 'Q,: for weighing, This method works wellwith refined fats or raw fats with FFA 3 percent and below, but fats withhigh FFA should be refilled ( see Note under A-l.2.1 ).

-The molecular mass of C P • calcula t~d on the basis of 2 moles of Itearic acid and'1 mole of the epoxy acid in one mole of triglvceride.

fMe.lured peak area uf the dillh 1 derivative is multiplied by a factor 0'7 which i.the ratio or molecular mass of dihydroxv ester to the disHyl derivative. This figure IIdivided by detector response factor, 0·95, to let the actual peak area of dihvdrox« ester.

:The molecular rna•• of ' Q.' calcula ted on the basil of 2 Jl)olcs of stearic aCId anelI mole of the dihydraxy acid in 1 mole of tric1yccridc.

lQ

II I 7375 • 1179

C 18( min)

•

C 16:0

C17:0

C 20:0

tiME IN MINUTES

Flo. 1 TYPICAL GLe SCAN

A-2.1 Procedure

A-2.1.1 Weigh accurately about 10 g of salseed fat and dissolvein 100 In} of hexane and sti.. with 20 g of silicic aciu. for 4 hours on amagnetic.stirrer. Filter the slurry using a Buchner funnel and distil the

·BDH laboratory reagent grade for lipid chromatography or any other silicic acidof equivalent grade activated at 110· for 1 hour before use may be used.

11

II • 7175• 1179

filtrate to obtain the normal triglyceride.. This fractinD does not showthe presence of 'P' or 'a: when 2 mg of the material is chromatographedon a TLC plate indicating that 'P' and 'a: are completely adsorbed( ", Note ).

NO'1'II - In case of silicic acid (rom other lOurCei. it II neceuary to do this TLCtelt to enlure absence of ' P , and • Q.' in the hexane extract and to arrive at theappropriate (at: .Uicic acid ratio. For jastanee a ratio of I : I wa. found suitablewith lilicic acid (rom allother source.

A4.1.2 Stir the residual silicic acid with 100 mJ of chloroform for1 hour and filter. Repeat the operation with a fresh lot of 100 ml ofchloroform. Distil the combined filtrate to obtain the enriched flaction•

.&-2.1.3 Dissolve a known mass ( 80 to 90 mg ) of the enriched fat in0-5 ml of chloroform and apply as a streak on a preparative plate of 1 mmthick silica geJ and develop four such plate. using hexane, ether and aceticacid ( 60 : 40 : 1 ) as a solvent system. Visualize the bands in an iodinechamber ( ", Fig. 2 for typical chromatogram) and scrape the bands atRr 0-S4 and 0·28 corresponding to 'P' and'a:. Transfer quantitatively intotwo separate thimbles and extract with chloroform in a Soxhlet, It takesabout I hour for extraction. After extraction, distil the chloroform andtransfer the residue carefully through a Whatman filter paper No. 42 ( orequivalent) to a tared 5 ml round-bottom flask using ether. Wash thefilter paper thoroughly with ether and collect the washings in the lameflask. Evaporate ether under a stream of nitrogen and weigh the flask toa constant mass by keeping it in an oven at 105°0.

A-2.2 CalcalatloDA-2.2.1 Calculate the amount of 'P' and ca: in the original fat from the

mals of the residue. Express the results as percentage of epoxy anddihydroxy fatty acids by dividing the sum of 'P' and 'Q: by 3 (s" alsoNotes 1 and 2 ).

NOT. 1 - The specified solvent system ( hexane: ether: acetic acid: : 60 : 40 : 1 )i. sati.factory for relolviol ' Q. t but the resolution of cpt (rom normal trillyeeridelis occasionally poor and appears to depend on the activity or lilica gel, The close Brvalue. of • P , and normal triglyceride. is then likely to lead to errors in eltimation of, P", In such casel a slilhtly lesl polar solvent system ( hexane: ether: acetic acid: :80 : 20 : 1 ) will lead to • lood resolution; C P 'and I Q,' should then be determinedleparately usial appropriate solveot systems.

NOTS 2 - Geaerally, the pattern or separation of various con.tituents in descendio.order of Re on the plate is al (ollowl:

a) Normal triglyceride.;b) , P ';c) FFA;d) I. 2-di,lyceride.:e) 1, 3-dillyceridcl;f) • Q, 'i aDdI> MODollycerides.

12

IS I 7375 • 1919

NORMALGLYCERIDES

p

FFA

f2222ZZzZZZ7Z2 ??zz2/.222ZZ<.)<Z>ZZZZ??ZZZt2Z;Z22 1 1 2 Z Z Z><Q

1:2 OG

T:3 DG

fZ2Z/Z222??2?2?2 Q

.............-----------------------.........-- MG....O1Z Z t t = = ZZ Ib

FlO. 2 A TYPICAL CHROMATOGRAM OP SALSBED FAT INHEXANE : ETHER : ACETIC ACID ( 60 : 40 : 1 )

13

203 91276496 27 165 36 172 83 20

22 10 836 98 328 12 7262808

32 27

INTERNATIONAL SYSTEM OF UNITS (51 UNITS)

8a•• U.Ia.

QUANTITr UNIT SYMBOL

Lcnltb metre m

l\ia.. lulogram kgTime second

FI~trlc eui f("nt arnpere A

Thermodynamic ke-lvm Kte mpei ature

Luminous intrnJuy candele cdAmount of substance molt mol

Supplemeota"J UDJt8

QnANrrrY t tN1'I ~Y"BOJ.

Plane allgle ••dian lad

Solid angle .terachal' sr

Derived Valta

QlTj.NrrJTY 1'''lIT S"Mnnl DR#IN ITION

Foree newton N 1\l - I kg.lnJ~t

Energy joule J J - 1 NmPower wan Vi t W ... J Ji'Flux wtbrr ws 1 Wb c== 1 V.I

Flux dcn.ity tesla T I T .. I \\'u/tn'Frequency Lcrt:z liz 1 Hz IZS I ("fl (.....1)

Electric conductance Il(",lt~en. S J s » I A/VElectromottve force volt V J V - ) WjAPressure. atresl pascal Pa J Pa .-e 1 Nitti'

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