specifications and criteria for identity and purity of

SPECIFICATIONS AND CRITERIA

FOR IDENTITY AND PURITY OF

SOME FLAVOURING SUBSTANCES

AND

NON-NUTRITIVE SWEETENING AGENTS

• Food end Agriculture Orgenizetlon of the United Netlona , World Heelth Orgenlzetlon

1969

FAO Nutrition Meetings

Report Series No. 448 WHO/FOOD ADD. / 69.31

SPECIFICATIONS AND CRITERIA FOR IDENTITY AND

PURITY OF SOME FLAVOURING SUBSTANCES

AND

NON-NUTRITIVE SWEETENING AGENTS

Issued jointly by FAO and WHO

The contents of this document are the result of the deliberations of the Joint FAO/WHO Expert Committee on Food Additives which met

in Geneva, 21-28 August 1967.L!_

• World Health Organization

Food and Agriculture Organization of the United Nations Rome 1969

L!_ Eleventh Report of the Joint FAO/WHO Expert Committee on Additives. FAO Nutrition Meetings Report Series, 1967, Wld Hlth Org, techn. Rep. Ser., 1967, 383.

Food No. 44_;

i

CONTENTS

Introduction

Flavouring Substances:

Trans-Anethole Benzaldehyde Benzyl Acetate Butyl Acetate Isoamyl Butyrate d-Carvone 1-Carvone Cinnamaldehyde Citral Citronellol (98%) Citronel lol (90"fa) De canal Di acetyl Ethyl Acetate Ethyl Butyrate Ethyl Formate Ethyl Heptanoate Ethyl Lactate Ethyl Laurate Ethyl Methylphenyl Glycidate Ethyl Nonanoate Ethyl i so-Valerate Ethyl Van ill in Eugenol Geranyl Acetate alpha-Ion one beta-Ion one Linal ol ( 9

9~)

Linal ol ( vr) Linalyl Acetate (9

9c:!;)

Linalyl Acetate ( .,-,..,) Maltol I-Menthol dl-Menthol Menthyl Anthrartilate Menthyl Phenylacetate Methyl Salicylate gamma-Nonalactone Nonanal Octanal Phenylacet.aldehyde Piperonal Y -Undecala otone Van ill in

Page 1

3 4 5 6 7 8 9

10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46

Non-Nutritive Sweeteners

Calcium Cyclamate Calcium Saccharin Cyclohexylsulfamic Acid Dulcin (1) Saccharin Sodium Cyclamate Sodium Saccharin

Annex - Methods of Analysis Test Solutions

ii

47 49 51 53 54 56 58 60 69

- 1 -

INTRODUCTI ON

The specifications and criteria for identity and purity of some flavouring substance• and non-nutritive sweetening agents contained in thi publication are the r esult of the deliberation• of the Joint FAO/WHO Expert Co11111ittee on Food Additives in their eleventh meeting held at Geneva on 21-28 August 1~67. Th••• should therefore be oonaidered together with the report of thia •••ting. 1/

While oonaidering the apecifioationa of th••• aubatanoea, the co .. itt•• had also agreed that :

l. Specific gravity should be given for 25°/25° a.a well aa f or 2fP/2fP where available.

2 . Other &dd.itioD&l criteria such aa moisture limits and diatillatioa raac• need not appear in the apecifioationa, but aa;r be aae:tul in oo.aeroial tranaaotioaa.

3. .uaa;ra uleaa an upper liait ia specified, there ia a iapliecl upper liait ot 100.~. .

4. hld.o trace el••taa 8peo1tio liaita nHcl DOt be ••t tor araeaio, lead. ucl otaer el•eata precipitable~ )vdropn aulplu.u uur aoicl oonditiou ia the oaae ot 'l;hoH flavouriq aaNuaoea 'l;hat an G.Uiilled., lleouae tll.eir abaeaoe ia ao proet ·~ poa ll&Dafaoturiq praouoe. The heavy .. iala liat ia iaolulllea ia 1;he cue of oryatalliHcl au'batanoea. It ia reoop1 .. 4l 'l;aat the flavouring aa'l;eriala are '8•cl 1D coaoeatrabou apreaaible ia parts per aillion (100) and theH toxic el .... ta aa;r be preaeat, it a'I; all, ia 'l;heae aateriala ia aiailar ooaoeatratioa!~ g1v1114r aet con'l;eata in 1;he food in the ruic• ot parts per 1~2 parts of food, uoua'l;a whioh would no'I; iaoreue aipiticuU7 aor be u-teo-table againat the •aria'ble llaok4rroua loacl of theae 'l;oxic trace el•ea-ta uturall7 preau1; in fooda.

5. Other ooapoaeniaa la geaeral, on 1;he baaia of aanufaoturiag pro••••••, the other ooapoaeata aaking up -the differeaoe be'l;weea tae aaaa;r figure aacl lC>OJ' aeea ao-t be aen'l;ionecl if 1;he7 are oouitlerecl 'l;o be of the••• paeral claaa aa 1;he aaterial apeoifiecl. Por ezaaple, in 'l;he cue of eaters, the otll.er ooapoaea'l;a are likely to be atartillg aoicl ancl alcohol ancl the higher ucl lower hoaologoaa ea-tera, aoida and. aloohola. la the oaae of alcle)vclaa, the oorreapoading acida aq be preaen'I; aa a result of oxidation. hrther liaita on iapuritiea nHcl to be specified only in the caae of auba-tancea where in the light of preaen-t ~ -technological practice this ia cleeaecl neoeaaary for 'l;oxi.cologioal reaaona.

!/ Eleventh Report of the Join-t FAO/WHO Expert Co11111ittee on Poocl Aciclitivea, PAO Butrition Keating Report Seri•• 44(1968)1 WHO Teoh. Report Seri•• 383(1968).

- 2 -

6. The identification of the flavours substances would not be specific if based only on the criteria of identity and purity detailed in this publication. The identity can be detel'lllined more specifically using additional indicative methods, such as gas liquid chromatography, infra-red spectrometry and nuclear magnetic resonance. When using such techniques for comparing the substance with its standard sample, it needs to be realized that many flavouring substances are not completely stable.

Azl¥ new information and co11111ents relating to the specifications of these products should be addressed to1 Food Standards, Additives and Regulations Section, Hutrition Division, FAO, Rome, Italy.

For detailed monographs on toxicological evaluation of these substances a reference is invited to the publication: "Toxicological .i:valuation of Some Flavouring Substances and Non-Hutritive Sweetening Agents." l/

l/ FAO Nutrition Meeting Report Series Ho. 44A; WHO/Food Add./68.33

3

JECFA/l/11/1967

TRANS-ANETHOLE

(trans-p-Propenylanisole)

Criteria

Refractive index, w20D 1.5570 - 1.5610

Specific gravity, 25°/25° 0.983 - 0.988

Solidification point Not below 20°

cia ia011er Not aore than (l~)*(by gaa-liquid chroaatograpq)

Alde~des and Ketones (a)

No Phenolic colour reaction (b)

Solubilit7 in ethanol 1 al dissolves in 2 al 9(1/, ethanol

(a) Alde~des and Ketones. Shake 10 al with 50 al of a aaturated solution of sodium biaulfite in a glass-stoppered, graduated c7linder, and allow the mixture to stand for six houra. The volume of the sample does not diainiah appreciabl7, and no crystalline deposit separate•.

(b) Phenols. Shake 1 ml with 20 ml of water, and allow the liquids to separate. Filter the water l&7er through a filter paper previously moistened with water, and to 10 ml of the filtrate add 3 drops of ferric chloride T.S. No purplish colour is produced.

* Tentative

4

JECFA/2/ll/1967

BENZALDEHYDE

(Benz oic Aldehyde )

Criteria

Assay Not lees than 97% (b) c7H60

Refractive index, N20D 1.5440 - 1.5470

Speciff c gravity, 25°/25° 1.041 - 1.046

20°/20° 1.043 - 1. 052

Chlorinated compounds Limit test (a)

{a) Chlorinated coapounli8. Prooeed as directed in the Annex.

(b) Asa~. Weigh accurately about 1 gram, and proceed aa direoted .in the Annex under Aldebydes, uaing 53.06 as the equivalence factor(~) in the calculation.

Criteria

5

BENZYL ACETATE

(Acetate de benzyl) Benzyl ethanoate

J EC FA/3 /ll/1967

Assay Not less than 97% (a)

C9Hl002

Refractive index, N20n

Specific gravity, 25°/25° 20°/20°

Acid Talue

Chlorinated compounds

1. 5010 - 1. 5040

1.052 - 1.056 1. 055 - 1. 059

1 ll&X

Limit test (b)

(a) Aaaq. Weigh accurately altout 900 ag, uul proceed u direoted. ill th• Allllex 'llllder Kater Dehrainatioll, uaing 75.10 u the equiTalello• !aotor (K) ill the oalculaticn. The reault• ahould be corrected tor the aoid value,

(b) Chlorinated ooapounda, Proceed aa directed ill the Allllex.

Criteria

Assay

20 Refractive index, N D

6

BUTYL AC:m'ATE

(Butyl ethanoate)

Not lees than 95% (a)

C6Hl202

1. 3920 - 1. 400

Specific gravity, 25°/25° 0.876 - 0.883 20°/20° 0.880 - o.887

Acid value 1 max

JECFA/4/11/1967

(a) AHq. Weigh aoourahl7 about l graa, anci proceed aa ciireoteci in the .Annex under Z•ter Determination, uainc 58.08 a• the equivalence faotor (e) in the calculation.

7

I SOAMYL BUTYRATE*

( Butyrate d'isoa myl ) Criteria

Refractiv e index, N20D 1, 4090 - 1,4140

Specific gravity, 25 °/ 25 ° 0 .860 - 0,8 64

200/ 200 0.863 - o.868

Acid value 1 max

Solubility in ethanol 1 ml diaaolvea in 4 ml 7<:J1, ethanol

JECFA/ 5 / ll/1967

(a) .la•q· V.ip aooura"el7 alloui l er•, aad. prooeed. u clireohd. in ihe .bmu 1U1d.er &.ier DeieninaUoa, uiac 79.13 u ihe equiY&leaoe fao"or (e) ia ihe oalnlaUoa.

• There i• ihe po••ibilii7 of aixiure• of i•o- IUld. nol'llal UliJl bui7raie.

8

. JECFA/ 6/11/1967

d-CARVONE

(+ -Carvone; d-l-Methyl-4-isopropenyl-6-cyclohexen-2-one)

Criteria

Assay

. d N20 Refractive in ex, D

Not leas than 95% (a)

C10H140

1. 4950 - 1. 5010


20°/20° 0.956 - 0.969

Angular rotation

Solubility in ethanol

+56° to +60°

l ml 5 ml

dissolves in 00,. ethanol

(a) Aaaq. Weigh aoouratal7 about l graa, and. prooead. aa cliraotad. in the Anna% Wilier ild.aqd.aa and X.ton••, 4.1 Jb'd.ro~luiD.• ••taocl, using 75.11 aa the aquivalanoa faotor (il) in the oaloulat1011; .2.£ alternatively by 4.2, Bautral Sulphite Method..

9

JECFA/7/11/1967

1-CARVONE

(- -Carvone; 1-1-Methyl-4-isopropenyl-6-cyc lohexen-2- one)

Criteria

Assay Not lees than 95% (a) C10H140



Angular rotation -55° to -62°

Solubility in ethanol 1 ml dissolves in 2 ml 7CJ/, ethanol

(a) Aaa8¥• Weigh accurately about l gram, and proceed aa directed in the Annex under Aldeh,ydea and Ketonea, 4.1 Jqdroxylamine Method, using 75.11 aa the equivalence factor (E) in the calculation.

Criteria

10

CINNAMALDEHYDE

(Cinnamic Aldehyde; Cinnamal)

(Ald6hyde Cinnamique)

JECFA/8/11/1967

Assay Not lees than 98% expressed as (b)

c9

H8o

d N2on · Refractive in ex,

S ' f ' 't 25°/25° peci ic gravi y,

Acid value

Chlorinated compounds

Bydrooarbone


1. 6190 - 1. 6250

1.046 - i.052

10 111ax

Limit test (o)

(a)

1 ml 5 ml

dissolves in &:Jfc. ethanol

(a) B7d.rooarlM>1u1. Keaaure lO al troa a pipe'\ in'\o a 100 al oaa•ia flaak and ad.d 75 11111 of a freahly prepared aolution of •odilUI bi•ulfite (12 in 100) preYiou•ly hea'\ed to a tempera'\ure of 85°. Shake the flaak vigoroualy until •olution ia complete, then add autficient •odiwa bisulfite solution to raise the meniacua within the graduated portion of the neck, Bo oil separates,

(b) Aas~. Weigh accurately about 1,5 grams, and proceed a• directed in the Annex under Aldehyde• using 66.08 aa the equivalence factor (K) in the oalculation,

(c) Chlorinated compounds. Proceed as directed in the Annex,

- 11 -

JECFA/9/11/1967

.rum ( 3, 7 Dimethyl-2, 6-ootadienal; a mixture of the geometric isomers neral and gera.nial)

Criteria

Ass&7 l'ot leas than 9~ e:i:preaaed as · ( a)

010Bi6°

Refraotive index, N20D 1.4850 - 1.4910

Speoifio gravity, 25°/25° o.883 - 0.891

20°/20° o.886 - 0.894

A.oid value

Liai ts of Impurities

As

Heavy aetals (as Pb)

Pb


5 au.

3 ag/kg au:

40 ag/kg IIU:

10 ag/k.g aax

1111 dissolves in 7 ml 7~ ethanol

( a) Ass&7. Weigh aoouratel7 about 1 gr1111, and prooeed as directed in the .Annex under ildehydea and Ketonea, 4.1 HydroX7lamine Method, using 76.12 as the equi valence taotor (E) in the oaloulation1 or alternatively b7 4,2, the Neutral Sulphite Method.

12

JECFA/10/11/1967

CITRONELLOL

(98% Total alcohols)

Criteria

Assay

Refractive index, N20D

Not less than 98% expressed as (a)

C10H200

1. 4530 - 1. 4580


Angular rotation

Ester value


-0°to +4°

2 max (b)

1 ml dissolves in 2 ml 7o% ethanol

{a) Aaaq. ProoHd. 1111 cliNote& 1a the .Amle% wader Total iloohol. Weigh aoovahl-7 altout l.2 gram• of the aoet7lated aloohOl for the •aponifioation, and Wle 78.ll aa the equiYalenoe factor CJ) ia the calculation.

(b) Bater, Weigh aoouratel7 altout 5 grama, and proceed as directed. in the Anne% under Eater Determination,

Criteria

Assay

CITRONELLOL

(90% Total alcohols)

JECFA/11/11/1967

Not less than 9r:tfo expressed as (a)

ClOH200

Refractive index, ~D 1.4520 - l.4620

Specific gravity, 25°/25° o.848 - o.860 20°/20° 0.853 - o.864

Allgular rotation

Eater Value


-5°to+5°

Not more than 2$, (b) calculated as citronellal (c1oHi80)

4 aax (o)

l ml dissolves in 2 .al 7afo e.thanol

(a) .uaq. Proceed. aa direohd. ill the .ums:it under Total .iloollol. Weigh &eOl&l"ateq uoat l.2 sr-• ot ihe aoe~latsd. aloohol tor the aaponifioation, and. uae 78.13 aa the equivalence taotor (B) in the oaloulation.

(b) .ilcle)Q'dee. Weigh aoouratel7 abo'llt 5 graae, and. proceed. .. directed. in the .umex under Ald.e)Q'd.ee and. Ketone•, 4.1 ]J.7droZ,Jluine Method., uaing 77 .13 a• the eCJ.Uivalenoo taotor (S) in the oalC'lllation tor oitronellal lC1oH1so) .

(o) &eter. Weigh aoouatel7 about 5 craa•, and. proceed. aa directed in the .ume:it under Ester Deteraination, using 99.15 as the eqv.ivalsnce factor (E) in the oalO'lllation tor citronellyl acetate (C12H2202).

14

JECFA/12/11/1967

DECANAL

(Aldehyde C-10; Capraldehyde, Decyl aldehyde)

(Ald~hyde caprique)

Criteria

Assay


Not lees than 9~ (a)

C10H200

1. 4270 1. 4350

Specific gravity, 25°/25° 0,821 - 0,835

Acid value 10 max

(a) .ua-.,, W•igh accu.ratel7 about 1.5 grams and proc•ed u directed in the AJmex under ild•b¥ciH, uaing 7a.14 aa the •quival•nc• factor (E) in the calculation.

15

JECFA/13/11/1967

DIACETYL

( 2, 3-Butanedione; Dimethyldiketone; Dimethylglyoxal) ( Diac~tyle)

Criteria

Assay

Rf t . . d N20 e rac ive in ex, D


C4Ht>°2

1. 3930 - 1. 3970

Specific gravity 25°/25° 0.975 - 0,990

Solidification point -1.0° to -4.0°

(a) .uaa;r. Weigb. accurahl; aDOut l graa, an4 prooeecl u direcied ia th• .&JI.nu: uder .ild•qd .. and Jr:etonH, 4,1 B;rdrolQ'l&ain• Kethod, uing 21,52 a• the •quivalence factor (K) in the calculation.

Criteria

Assay

Refractive index, N20

D

16

JECFA/14/11/1967

ETHYL ACETATE

(Acetate d'ethyle)

Not lees than 97% (a) expressed as

C4H802

1. 3710 - 1. 376o

Specific gravity, 25°/25° 0.894 - 0.901 20°/20° 0.897 - 0.906

(a) Assay. Transfer about 1.5 grams, accurately weighed in a tared, stoppered weighing bottle, to a suitable flask, add 50.0 ml of 0.5 N sodium hydroxide, and heat on a steam bath under a reflux condenser for 1 hour. Allow it to cool, add phenolphthalein T.S. and titrate the excess sodium hydroxide with 0.5 N hydrochloric acid. Perform a blank determination, and make any necessary correction. Each ml of 0.5 N sodium hydroxide is equivalent to 44.06 mg. of C4Ha02•

Criteria

17

ETHYL BUTYRATE

(Butyrate d'~tbyle)

JECFA/15/11/1967

Assay Not lees than 97% expressed as (a)

C6H12°2


D 1. 3900 - 1. 3950


Acid value 1 max

(a) AaaiQ". Weigh accurately about l gram, and proceed aa directed in the .Annex under Kater Detel'lllination, uaing 58.o8 aa the equivalence factor (E) in the calculation.

Criteria

Assay


18

JECFA/16/11/1967

ETHYL FORMATE


C3H602

1. 3580 - 1.3630


Acid value 2 max

(a) A••~· Weigh accurately about 500 mg, and proceed a. direoted in the .Annex under Xater Determination, uaing 37.04 as the equivalence faotor (E) in the calculation. Modify the procedure by allowing the mixture to atand at room temperature for 15 minutea, inatead of heating on a steam bath for l hour.

Criteria

Assay

19

JECFA/17/11/1967

ETHYL HEPTANOATE

(Ethyl Heptoate, Ethyl Oenanthate)

(Oenanthate d'~thyle)

Not less than 97% expressed as (a) C9H1802

Refractive index, N20D 1. 4110 - 1. 4160

Specific gravity, 25°/25° 0.864 - 0.872 20°/20°

Acid value l max

(a) A8•&¥• Weigh accuratel7 about l graa, anli prooHd aa direoted in the Annex under Bater Determination , uaing 79.12 aa the equivalence faotor (E) in the oaloulation.

20

JECFA/18/11/1967

ETHYL LACTATE

(Ethyl 2-Ji¥droxypropionate)

Criteria

Assay Not less than 98% expressed as (a)

C58io03


Speoifio gravity, 25°/25° 1.028 - 1.034 20°/20° 1.031 - 1.037

Aoid value 2 max

(a) AaH3. Weigh aoouratel7 about 700 1118, and procHd aa directed in the Annex UDder later Determination, using 59.07 a• the equivalence factor (E) in the calculation.

Cri teria

Aeeay

21

ETHYL LAURATE

( Ethyl Dodecanoate)

JECFA/19/11/ 1967

Not leee than 98% expressed ae (a)

C14H28°2


Specific gravity, 25°/25° 0.858 - 0.8 62

Acid value

(a ) Aaaq. Weigh aoouratel~ about 1.5 graaa, and. proceed aa clirected in t he Annex under Kater Detel'lllination, ueing 114 .2 as t he equivalence factor (E) in the calculati on.

22

JECFA/20/11/1967

ETHYL METHYLPHENYL GLYCIDATE

(Aldehyde C-16*)

Criteria


C12H1403


D (1.5040 - 1.5100)**

Specific gravity, 25°/25° (1.086 - 1.092)**

Chlorinated compounds Limit test (b)

Acid value 3 max

(a) Aaaa3. Weigh accurately about l graa, and procHd aa directed in the Annex under Kater Determination, uaing 103.l aa the equivalence factor {E) in the calculation.

(b) Chlorinated compounde. Prooeed aa directed in the Annex.

* Miscalled, but to be indexed under this name as well.

** Tentative

Criteria

23

ETHYL NONANOATE

(Ethyl pelargonate)

(P~largonate d'etbyle)

JECFA/21/11/1967

Assay Not less than 98% expressed as (a) C11H22°2

Refractive index, N20 D

1. 4200 - 1. 4260

Specific gravity, 25°/25° o.863 - o.867

Acid value 3 max

(a ) AH,q. Weigh accura1ie~ allou1i 1.5 grams, and procHd aa direc1ied in 1ihe Anne% 1U1der Ba1ier De1ieraina1iion , uai114r 93.15 aa 1ihe equivalence fao1ior (E) in 1ihe calculation.

Criteria

Assay

20 Refractive index, N D

24

ETHYL iso-VALERATE

(Ethyl 2-Methylbutyrate)

(Isoval~r~te d'ethyle)

JECFA/22/11/1967

Jot l••• than 9&.' and not •or• than 101~ (a) 9%1)r•aaed aa c7a14o2

1.3930 - 1.3990

Specific gravity, 25°/25° o.862 - o.866 20°/20° o.866 - 0.871

Aoid value 2 m.&X

(a) Aaaa,y. Weigh accurately about 1.5 grama, and proceed as direoted in the Annex under Ester Determination, uaiJ18 65.10 aa the equivalence factor 1(E) in the calculation.

DHcription

A. Solubilit;y:

25

J"i.CFA/ 23/11/1967

ETHYL VANILLili

(3-Ethox;y-4-h,Ycirox;ybenzaldehyde)

(Ald.4h,Yd.e eth,Ylprotocatachnique)

Fine, white or slightly yellowish crystals, having a strong vanilla-like odour and taste; affected by light .

Identification Testa

Water: Insoluble 6(},\, Ethanol a l g in 10 111.

B. Wara about 100 ag. of the aaaple with l ml of 25 percent h,Ycirochloric acid until ooaplete solution is effected.. Cool, add. about l al of lQ'd.rogen peroxide T.s., and allow the aixture to stand, with frequent shaking, until preoipitation ia complete (10 to 20 min.), Add an equal nluae of benae11e and. shake thoroughly. The benze11e l&,1er ia violet in colour {di.atinction from vanilla).

C. The aaaple ia extracted. co•pletely from solution by aha.kizac with a saturated. solution of sodium biaulfite , froa which it is precipitated. by acid.a.

D, Add lead. aubacetate T.S. to a cold solution of the sample. A white precipitate which ia sparingly soluble in hot water, but soluble in acetic acid, is produced.

Limits of Impurities

Heavy metals: (as Pb)

Not more than 10 mg/kg

26

J ECFA/24/11/1967

EUGENOL

(4-Allyl-2- methoxyphenol; 4-Allylguaiacol)

Criteria

Assay


D

Speoifio gravity, 25°/25° 20°/20°

H,ydrooarbona

Phenol

Not leas than 98% by volume, of phenols (a) as eugenol (C1oH12o2 )

1.5380 - 1.5420

1.064 - 1.070 1.065 - 1.071

(b)

(o)

(a) Assay. Proceed as directed in the Annex under Phenols.

(b) H,ydrooarbons. Dissolve l ml in 20 ml of 0.5 N sodium }Q'droxide in a stoppered 50 ml tube, add 18 ml of water, and mix. A clear mixture results immediately, but it ma.y become turbid when exposed to air.

(o) Phenol. Shake l ml with 20 ml of water, filter, and add l drop of ferric chloride T.S. to 5 ml of the clear filtrate. The mixture exhibits a transient gra.yish green colour, but not a blue or violet colour.

27

JECFA/25/11/1967

GERANYL ACETATE

(2,6-Dimetbyl-1,6-octadien-8-yl Acetate)

Criteria

Assay


Not leas than 90% of total eaters, (a) calculated as geranyl acetate

( Cl2H2002)

1. 4570 - 1. 4640

Specific gravity, 25°/25° 0.896 - 0,914

Angular rotation


Acid value

0 0 -2.0 to +3,0

1 ml dissolves in 4 ml 80% ethanol

1 max

(a) ABB83', Weigh accurately about l graa, and proceed. u d.ireoted. in the Annex under Kater Deter11ination, uaing 98.15 aa the equivalence factor {E) in the calculation.

Criteria

Assay


D

Specific gravity, 25°/25° 200;200

28

JECFA/26/11/1967

alpha-IONONE*

Hot leaa th&ll 95~ ketonea expreaaed aa (a) c13H20o

1.4970 - 1.5025

0.927 - 0.933 (0.930 - 0.934)**

(a) Aaaa.,. Weigh aoourately about 1.3 graaa, and proceed aa directed in the Annex under Aldehydes and Kstonea, 4.1 ~dro~laain• Method, using 96.15 aa the equivalence factor (E) in the oalculation.

* The product of 001111eroe used in food under this name ma;y contain varying proportions of isomeric iononea.

** Tentative

Criteria

Assay


29

JECFA/27 /11/1967

beta-IONONE*

lot leaa than 95~ ketone• expreaaed aa (a) c13B2o()

l. 5070 - l. 5230


(a) baq. Weigh aoouratel7 aboa'i 1.3 grams, and proceed aa d.ireo'iecl in the .lnnex under Alcle~dea and Ke'icnea, 4.1 ll7dro:icylamine Method, v.aing 96.15 aa the equivalence factor (E) in the calculation.

* The product of 0011111erce used in food under this name may contain varying proportions of iaoaeric ionones.

30

JECFA/28/ 11/ 1967

LINALOL, 95%

(Linal ool; 3 , 7-Dimethyl-1,6-octa dien-3-ol )

Criteria


C10H180

20 Refractive index, N D (1.4600 - 1.4640) *

Specific gravity, 25°/25° (0.858 - 0.867)*

Angular rotation (-2°to +2°)*

Ester value 1.5 max (b)

Solubility in ethanol 1 ml dissolves in 4 ml 60% ethanol to form a clear solution.

(a) Aaaa.,. Proceed as directed in the Annex under Linalool Determination, using about 1.2 grams of aoetylized oil, accurately weighed.

(b) Eaters. Weigh accurately about 10 grams, and proceed as directed in the Annex un~er Ester Determination.

* Tentative

31

JECFA/29/11/1967

LINALOL, 90%

(Li nalool; 3,7-Dime t byl-l,6-octadien- 3-ol )

Cri teria

Assay

Refract i ve i ndex, x20D

Not less than 9o% expressed as (a)

C10H180

1.4604 - 1.4655

Specific gravity , 25°/25° 0.858 - o.868

Es t e r value

Solubi lity in e t hanol

4 max (b)

1 Ill. dissolves in 4 Ill. fl:II, ethanol to fora a clear solution.

(a) Aaaay. Proceed aa directed. in the Annex Wld.er Linalool Determination using about 1.2 gram• of acetyliaed. oil, accurately weighed .

(b) Estera. Weigh accurately about 10 grams, and proceed as directed in the Annex Wlder Eater Determination.

32

JECFA/30/11/1967

LINALYL ACETATE, 90%

(3,7-Dimethyl, 1,6-octadien-3-yl Acetate)

Criteria

Assay


Not less than 90% of esters, (a) calculated as linalyl acetate

( Cl2H2002)

1. 4490 - 1. 4580 ,


Acid value


2 max

1 ml 5 ml

dissolves in 70% ethanol

(a) Aaaiq. Weigh accUl"ately about l gram, and proceed as directed in the Annex under Ester Determination, using 98.15 as the equivalence factor (E) in the calculation. The volume of 0.5 N alcoholic potassium hydroxide consumed by the sample should be corrected for the Acid Value.

Criteria

Assay

33

JECFA/31/11/1967

LINALYL ACETATE, 9fF/o

(3,7-Dimethyl,l,6-octadien-3-yl Acetate)

lot less than 9&f, of esters, (a) calculated as linalyl acetate ( C12H2002)

Refractive index, 120

D 1. 4490 - 1. 4530

. 25°/25° 0.895 - 0.908 Specific graTity, 20°/20° (0.899 - 0.906)*

Aoid Talue 1 au:

(a) Aeeay. Weigh accurately about l graa, and proceed as directed in the Annex under Eater Determination, using 98.15 a• the equivalence factor (e) in the oalculation. The volume of 0.5 H alcoholic potassium hydroxide consumed by the sample should be corrected for the Acid Value.

* Tentative

.34

JECFA/32/11/1967

~

(3-Hydro:xy-2-methyl-4-pyrone)

Criteria

Assay Not less than 98% (a)

C6H603

Melting range

(a) Assay: Standard Solution. Weigh accurately about 50 mg. of F.C.C. Maltol Reference Standard, di&solve it in sufficient O.l N hydrochloric acid to make 250.0 ml , and mix , Transfer 5.0 ml of this solution into a 100 ml volumetric flask, dilute to volume wi th 0,1 N hydrochloric acid, and mix.

AS88J' Solution, Weigh accurately about 50 mg. of the sample, dissolve it in sufficient O.l N hydrochloric acid to make 250.0 ml, and mix. Transfer 5.0 ml of this solution into a 100 ml volumetric flask, dilute to volume with O.l N hydrochloric acid, and mix,

Procedure , Determine the absorbauce of each solution in al cm, quartz cell at the wavelength of maximum absorption at about 274 mu, with a suitable spectrophotometer, using 0,1 N hydrochloric acid as the blank. Calculate the quantity, in mg,, of c6n6o3 in the sample taken by the formula 5C(Au/As), in wfiich C is the concentration, in mcg. per ml, of F,C.C. Maltol Reference Standard in the Standard Solution , Au is the abeorbance of the Assay Solution, and As is the absorbance of the Standard Solution,

35

1-JlEli'l'HOL

(1-3-p-Nenthanol)

Criteria

Melting range

0 Specific rotation, (oq 2i

41 ° - 44°

JECFA/33/11/1967

36

dl-MENTHOL

(d,1-3-p-Mentbanol) Criteria

Melting range

Solidification behaviour

(-''2n50 Specific rotation, """

Readily oxidizable substances

(a)

(b)

JECFA/34/11/1967

(a) "SoliditioahOD bellaviour ot cll-aenthol. DehZ'lllin• aa directed in the ADnex, u•ing a •ample previou•ly dried in a deaioator over silica gel for 24 houra and adjuating the temperature of the cooling bath to a temperature b•tween 23° and 25°. dl-Kenthol solidifies between 27° and 26° . Continue the stirring. After a few minutes the temperature quickly rieea to 30.5° to 32°. 11

(b) Readily oxidisable substances in ell-menthol. Transfer 500 mg. of dl-menthol into a clean , dry test tube, add 10 ml of potassium permanganate with water to 100 al. Place the test tube in a beaker of water maintained between 45° and 50°. At 3~aecond intervals, quickly remove the test tube from the bath and shake. The colour of potassium permanganate is still apparent after 5 minutes.

Criter ia

31

MENTHYL ANTHRANILATE

(Me t hyl 2-Aminobenzoa t e)

(Anthranilate de M~thyle)

JECFA/35/11/1967

Assay Not less than 98% expressed as (a) c8H~02

Refract ive index, N20

D 1. 5800 - 1 . 5850


Solidification point liot less than 22°

Acid value 1 mai

(a) Aes53. Weigh accurately about l gram, and proceed as directed in the Annex under E1ter Determination, using 75.59 as the equivalence factor (E) in the calculation.

38

J ECFA/36/11/1 967

METHYL PHENYLACETATE

(Ph~nylacetate de M~ t hyle)

Criteria


C9H1002


Specific gravity , 25°/25° 1.061 - 0. 0 67

Acid value 1 max

Chlorinated compounds Limit test (b)

(a) Aasa,y. Weigh accurately about l gram, and proceed as directed in the Annex under Ester Determination, uaing 75.09 as the equivalence factor (E) in the calculation.

(b) Chlorinated compounds, Proceed as directed in the Annex,

Criteria

Assay

39

METHYL SALICYLATE

(Wintergreen oil, artifioial)

98% expressed as (a)

C8H803


D 1.5340 - 1. 5380

f t 25°/25° 1.180 to 1.188 Speci io gravi y,

JECFA/37/11/1967

(a) Ass~. Proce~d as directed in the Annex under Eater Determination, using about 2 graaa, accurately weighed. Use 50 ml of 0.5 I alcoholic potassium hydroxide and reflux on the steam bath for 2 hours. Each ml ot 0.5 N potassium hydroxide consumed is equivalent to 76.08 mg of c8H8o

3•

Criteria

40

gamma-NOBALACTONE

(1-Nonalactone)*

JECFA/38/11/1967


C9H1602


Speoifio gravity, 25°/25° 0.957 - 0.968

20°/20° 0.965 - 0.970

Aoid value 5 max

(a} Aaaa,y. Weigh accurately about l gram, and proceed aa directed in the .Annex under Eater Determination, using 78.ll aa the equivalence factor (E) in the calculation. Correct the number of ml of 0.5 N aloohclic potaaaium b¥droxide conaU111ed in the aaponifioation for the aoid value.

* Miscalled Aldeb¥de C-18, but cross-indexed under this name.

41

JECFA/39/11/1967

NON ANAL

(Pelargonic Aldehyde; Aldehyde C-9)

(Ald,hyde Pelargonique)

Criteria

Assay Not less than 9'2'fo expressed as (a)

C9H180


D 1.4220 - 1.4300

Specific gravity, 25°/25° 0.820 - o.830

Acid value 10 aax

(a) Aaaa.y. Weigh accurately about 1.5 grams, and proceed aa directed in the Annex under Aldehyde&, using 71.12 as the equivalence factor (E) in the calculation.

Criteria

OCT ANAL

(Caprylic Aldehyde; Aldehyde C-8)

(Ald6byde Caprylique)

,JE:'.. A/40/11/1967

Assay Not less than 9'i!/c expressed as (a)

CsH160


D 1. 4170 - 1. 426o

Specific gravity, 25°/25° o.817 - 0.830 20°/20° 0.820 - o.837

Acid value 10 max

(a) A.asay , Weigh accurately auout 1.5 grams, and proceed aa directed in the Annex under Aldehydes, using 64.11 as t he equivalence factor (EJ in the calculation.

43

JECFA/ 41/11/1967

PHENYLACETALDEHYDE

~-Toluic Aldehyde)

(Ald6hydect'-tolnique1 Ald6hyde ph6nylacetique)

Criteria

Assay


Not less than 90%* expressed as (a)

C8lia0

1.5240 - 1.5330


Chlorinated compounds Limit test** (b)

Aoid value 5 lll&JI:

{a) Aae~. Weigh accurately about l graa, ancl prooHa aa d.irectec;l. in the Annex unc;l.er ild.elqc;l.H , uaing 60.08 aa the equivalence factor (E) in the calculation.

(b) Chlorinatec;J. co11pounu: .la i n Annu.

* Caution: Because this compound ia unstable, it is not stored and used in this fora, but in solution, e.g. 5~ in ethanol.

** TeJltative

44

J EC?A/42 /11/1967

PIPERONAL

(Heliotropine; Piperonyl Aldehyde)

Criteria

Assay

Solidification or melting point

Heavy metals (as Pb)

Not less than 9()% expressed as (a)

C8H603

Not less than 35°

40 mg/kg max

(a) Assay. Weigh accurately about 1.5 grams, and proceed as directed in the Annex under Aldehyde•, using 75.07 as the equivalence factor (E) in the calculation.

Criteria

Assay

Refraotive index, N20

D

45

JECFA/43/11/1967

Y-UNDECALACTONE*

Not lees than 97% expressed as · (a)

CllH2002

1. 4500 - 1. 4550

Specifio gravity, 25°/25° 0.940 - 0.945 20°/20° 0.945 - 0.950

Acid value 5 max

(a) .Aa11a,y. Weigh accuratel7 about l gram, and. proceed &11

d.irected. in the Annex und.er Bater Determination, uaing 92.14 aa the equivalence factor (E) in the calculation.

* Miacalled. Aldehyde C-14, but to be cross-indexed under thie name.

Criteria

Assay

Melting range

Heavy metals (as Pb)

46

JECFA/44/11/1967

VANILLIN

(4-Hydroxy-3-methoxybenzaldehyde)

(Aldehyde methylprotocatechuique)

97 - 103% expressed as c8n8o3 (a)

Between 80.8° and 83°


(a) Assa.,. Standard Solution. Transfer about 100 mg, accurately weighed of Vanillin Reference Standard into a 250 ml volwaetric flask, add methanol to volwae, and mix. Transfer 2.0 ml of this solution into a 100 ml volumetric flask, dilute to volume with methanol, and mix,

Assa., ~olution, Transfer about 100 mg., accurately weighed, of the sample into a 250 ml volumetric flask, add methanol to volwae and mix. Transfer 2.0 ml of this aolution into a 100 ml voluaetric flask, dilute to volume with methanol, and mix,

Procedure. Determine the abaorbanoe of each solution in al om. qu.artz cell at the wavelength of maximum abaorption at about 308 mu, with a suitable spectrophotometer, using methanol as the blank. Calculate the qu.antity, in mg, of CsHs03 in the sample ta.ken by the formula 12.5c x (Au/As), in which C is the concentration, in mcg, per ml, of Vanillin Reference Standard in the Standard Solution, Au is the absorbance of the Asaa,y Solution, and A8 is the absorbance of the Standard Solution,

Chemical Bame

Empirical Formula

Molecular Weight

Definitiop

Deacription

A. Solubility:

47

CALCIUJl CYCLAMATE

(Cycl .. ate de oaloiua)

JECFA/ 45/11/1967

Calcium Cyclohexanesulfaaate, Calcium CyoloheJcylaulfaaate

C12H24Call206S2.H20

432.57

Calcium oyolaaate contain& not leas than 98 percent and not 1110re than the equiyalent of 101 percent of c12H24call206s2 ca~culated oa the dr1 buia, and conforms to the following apeoificationa.

White ociourleaa 017atala or oeyatalline powcier.

Hon-nutritive aweetener .

Ideptifioaliop Teata

Water: Soluble Ethanol1 Sparingl.7 soluble

B. To 10 ml of a ii solution add l al of hydrochloric acid, mix, add l al of bariua chloride T.S. The solution remaina clear, but upon the addition of l •l of a lo.' sodiua nitrite solution, a white precipitate ia foraed.

c. Al in 100 aolution givea poaitive teats for calcium.

Calcium Cyolaaate (cont'd) Jl!:CFA/ 45/11/1967

Loaa on drying:

Heavy metals (as Pb):

Selenium:

Cyclohe~lamine:

Purity Teats

Between 6 percent and. 9 percent when dried at 140° for 2 hours.



Not more than 100 mg/kg. (Method to be established).

Dissolve about 400 mg of a11111ple which has been accurately weighed in a mixture of 50 ml of water and 5 ml of hydrochloric acid, T.S. and titrate with O.l K sodium nitrite. Auu the last ml of titrant dropwiee until a blue colour is produced i111111ediately whsn a glass rod dipped into the titrated solution is streaked on a piece of ataroh iodide teat paper or alternatively the end point may be detected electrometrioally. When the titration is complete, the end-point is reproducible after the mixture hae been allowed to stand. for l minute. Each ml of O.l K sodi11111 nitrite is equivalent to 19.83 mg of c12u24ca H206s2 , calculated on the dry basis.

ca .. 10,1 xaae

ilapirical Poraula

Kolaoular Weight

Dffini Uon

J>eecripUoa

A. Solubility I

49

J~CFA/46/11/1967

CAl,ClU)( §ACClWilli

Calci\lll o-Benso•ulfiaid.e , CalciWD salt of 2,3-clihyciro-3-oxobenzi•o•ulfasol e

C14H5Ca1J206S2•

467.49

Calciua aaccharin cont ai ns no t leas than 95 percent of c14s8caJJ2o6s2 calculat ed. on

anhTdroua baaia and. conf'orma t o the following apeci ficationa .

White 017atala or a whi te ceyatalline powo.er, which i• odourlesa or haa a faint, aroaat ic odour and. an intenaely s weet taate even in di lute aol utiona.

Bon-nutritive aweetener.

Id.apti fioaSiop Teet•

Water1 ithanol1

l'rHly aoluble Soluble

B. Diaaolve about 100 mg of the aample in 5 ml of 5 percent aod.ium hydroxide solution, evaporate to deyneaa and gently fuae the reaid.ue over a small flame until it no longer evolve• a111111onia. After the reaidue haa cooled., dissolve it in 20 ml of water, neutralise the solution with diluted hydrochloric acid T.S. and filter. Th• addition of a drop of ferric chloride T.S. to the filtrate produce• a violet colour.

c. Mix 20 mg of the sample with 40 mg of resorcinol, add 10 drops of sulphuric acid and heat the mixture in a liquid. bath at 2ooO for 3 minutes. After cooling, add. 10 ml of water and an exoeaa of •od.iwn hydroxide T.S. A fluorescent green liquid re•ults.

50

Calciwn Saccharin (cont'd) JECFA/46/11/1967

D, Al in 10 solution gives positive test for calciwn,

E, To 10 ml of al in 10 solution add l ml of hydrochloric acid, A crystalline precipitate of saccharin is foraed, Wash the precipitate well vith cold water and dry at 105° for 2 hours, It melts between 226° and 230°.

Water:

Purity Teats

Not more than 15 percent as determined by Karl Fischer Titrillletre Kethod,

HeaY1 metals (as Pb): Not more than 10 mg/k8

Seleni1111: Not more than 30 mg/k&,

Weigh accurately about 700 mg of the sample and transfer it quantitatively to a lon&""neoked flask, having a capacity of 200 ml, and add 10 ml of a 30 percent w/v solution of sodiWII hydroxide in water; boil over a small flame for two minutes, avoiding loss by evaporation; cool, add 15 ml of hydrochloric acid, and boil again for fifty minutes under a reflux condenser, Cool, rinse the condenser with 50 ml of water, and pass a current of air through the flask to remove acid vapour. Connect with an ammonia distillation apparatus, add 20 ml of a 30 percent w/v solution of sodiwn hydroxide in water , and distil the ammonia into 40 ml of N/10 sulphuric acid; titrate the excess of acid with N/ 10 sodium hydroxide, using solution of methyl red as indicator. Each ml of N/10 sulphuric aoid , neutralized by the ammonia i n the distillate is equivalent to 20,22 mg of C14H8caN2o6s2•

51

JECFA/47/11/1967

CYCLOHEXILSULF.AJlIC ACID

(Aoide cyclohe:xylaulfanique)

SynonY11a Cyclohexaneaulfanic aoid

Empirical lol'llula c6a13103s

Siruoiur&l Formula

Koleoular Weight

Defi nijiop

Deacri ption

A. Solubility:

179.24

C)'clohe:xylaulfamic aoid ccniaina noi leaa ihan 98 per cent and noi •ore ihan the equivalent of 102 peroeni of C68i)I03s, calculated on the anh,ydJ'oua baaia and oonfol'lla io the following apecifioaiiona.

A pr actically colourl • • •, white c17atallin8 powder with a aw•• t-aour i aaie.

Hcn-nuiriiiY• aweetener.

I dentificat ion T••t•

Waier: Ji:ihanol:

Soluble Soluble

B. Aciaify a 2 percent aoluiion wiih hydrochloric acid, add l ml of barium chloride T,S. and filt er if any haze or precipitate fol'lla. To the clear aoluiion add l al of a 10 percent aoluiion of sodium nitrite. A whiie precipiiate for11a .

52

CycloheJQ-lsulfamic Acid (cont'd) JECFA/47/11/1967

LOH on cirying:

Heavy metals (a• Pb):

Seleniua:

C;yoloheJcylamine:

Purity THtS

Hot more than l percent on drying at 105° for one hour.

Hot more than 10 mg/kg.

Hot more than 30 mg/k&•

Hot more than 100 mg/k& (method to be eaiablished).

Tran•fer about 350 mg, accurately weighed, into a 250 ml flask. Diasolve the sample in 50 ml of water, add phenolphthalein T.S., and titrate with O.l H sodium hydroxide. li:ach al of O.l B •odiwa hydroxide is equivalent to 17.82 mg of c6H13Ho3s.

53

JECFA/48/11/1967

1m!&ll (l)

SJpOBJ118 Sucrol, Valsin

C8!f1ical 1... 4-ortho~a•n;rlurea

ilapiriqal PorpJlla c~20~2

Koleqular Weicht 180.21

Ptt9£ip$ioa Colourl••• or whit• cryatals, or a white cryatallin• powder, which ia odourleaa and haa a very aweet taste which ia appreciable even after a 3~fold dilutioa.

ll!!.!. Hon-nutritive aweetener.

A. SolubiU.t;r:

l deptifi oation Testa

Water: ~thanol:

Insoluble Soluble

B. On heating a sa11pl• in a teat-tube it decomposes with the evolution of 1111111onia (odour ), and produces a white subli•ate in the cool portion of the teat-tube.

c. To 20 mg of the sample add 4 drops of liquified phenol and 4 drops of sulphuric aoid, and heat to boil. Cool and carefully superimpose 3 ml of potasaium hydroxide T.S. over this mixture. A blue colour appears at the plane of contact in 2 to 3 minutes.

Chemical Name

lwllpirical Formula

Structural Formula

Molecular Weight

Definiiion

Description

54

JECFA/49/ll/1967

SACCHARIN

(Saccharin•)

o-Benzosulfiaide, 2 1 3-dihydro-3-oxobenzisosulfonazole

0

II

183.19

NH

Saccharin contains not less than 98 percent of c7a5xo3s after drying and con-

forms to th• following specifi cations.

White crystals or a white, crystalline powder, odourless, or with a faint , aromatic odour.

Non-nutritive sweetener.

Identification Tests

A. Solubi lity: Water: Sli.ghtly soluble , but soluble in basic solutions.

Ethanol: Siaringly soluble

B. Dissolve about 100 mg in 5 ml of 5 percent sodium hydroxide solution, evaporate to dryness and gently fuse the residue over a small flame until it no longer evolves ammonia. After the residue has cooled , dissolve it in 20 ml of water, neutralize the solution with diluted hydrochloric acid T.S. and filter. The addition of a drop of ferric chloride T.S. to the filtrate produces a violet colour.

55

Saccharin (cont'd) JECFA/49/11/1967

c. Mix 20 mg with 40 mg of reaorcinol, add 10 drops of aulghuric acid, and heat the mixture in a liquid bath at 200 for 3 minutes. After cooling, add 10 ml of water and an excess of sodium hydroxide T.S. A fluorescent green liquid is produced.

D. Melting range: Between 226° and 230°.

Losa on drying:

Heavy metal• (aa Pb):

Selenium:

Purity Teats

lot more than l percent.

Not more than 20 mg/k8·

Not more than 30 mg/k8·

Transfer about 700 mg previoualy dried at 105° for 2 hours and accurately weighed, to a long-necked flask, having a capacity of 200 ml, and add. 10 ml of a 30 percent w/v aolution of sodium hydroxide in water; boil over a small flame for two minutes, avoiding loss by evaporation; cool, add 15 ml of hydrochloric acid, and boil again for fifty minute• under a reflux condenser. Cool, rinse the condenaer with 50 ml of water, and pass a current of air through the flaak to remove acid vapour. Connect .with an &111111onia distillation apparatus, add 20 ml of a 30 percent w/v solution of sodium hydroxide in water, and distil the ammonia into 40 ml of N/10 sulphuric acid; titrate the exceea of acid with 1/10 aodiua hydroxide, using solution of methyl red as indicator. Each ml of N/10 sulphuric acid, neutralized by the ammonia in the distillate,ia equivalent to 18.32 mg of C7H5No3s.

Chemical Name

Empirical Formula

Structural Formula

Molecular Weight

Definition

Description

A, Solubility:

Jb.CFA/50/11/l':}67

SODIUM CYCLAMATE

(Cyclamate de sodium)

Sodium Cyclohexaneaulfamate, Sodium Cyolohexylaulfamate

NH-S02-0Na

201.22

Sodium cyclamate contains not leas than 98 percent and not more than the equivalent of 101 percent of C6H12NNao3s and conforms to the f ollowing specifi cations.

White, odourless crystals or crystalline powaer which in dilute solution is about 30 times aa sweet as sucrose.


Identification Teats

Water: Soluble Ethanol: Practical l y i nsoluble.

B, To 10 ml of al~ so lution add 1 ml of hydrochlo r i c acid, mix , then add l ml of barium chloride T ,S, The so l ution remains clear but upon the addition of 1 ml of sodium nitrite solution (1~ ) a white pr ecipitate forms,

C. Al~ solution gives positive t ests f or aouium,

57

Sodium Cyclamate (cont'd) J ECFA/50/11/ 1967

PUfib THh

LOH on d.J7ing1 ~~;0 m;:: !!:"h~u!'.~rcent

when dried at

Heavy metal• (a• Pb): Hot more than 10 mg/kg .

Seleniua: Not more than 30 mg/kg.

Cyclohe:iql aaine: Hot more than 100 mg/kg (method to be Htabliahed.).

Diaaolve about 400 ag, preYioualy dried at 105° tor l hour and. accurately weighed., in a aixtar. of 50 al of water and 5 al of hyd.roohloric acid. , T.s. and. titrate with 0.1 K aod.iwa nitrite • .Add. the laat Ill of titraat d.ropwiae until a blue colour i• prod.uced. iaaed.iately whea a glaaa rod. dipped. into the titrated. aolution ia atreaked. oa a piece of starch iod.ide teat paper or alternatively, the end. point a~ be detected electroaetrioally. llhen the titration ia complete, the end. point ia reprod.uoible after the aixture haa been allowed. to stand for l ainute . li:ach al of 0.1 • aod.iwa nitrite ia equiYalent to 20.12 ag. of C6H12'-Ha03s.

JECFA/51/11/1967

SODIUK SACCHARIN

(Sel de aodium de la saccharine)

Che111ioal 11-•

Empirical Formula

Molecular Weight

Definitiop

Description

A. Solubility :

Sodium o-Bensoaulfimide, Sodium aalt of 2,3-dip~dro-a-oxobenzisosulfazole

c7H4NNao3s .2H20

241.20

Sodium saccharin contains not leas than 98 percent and not more than the equivalent of 101 percent of c7H4.liNa03s (205.17) after drying and oonforma to the following specifications.

White crystals or a white, crystalline, effloresoent powder which is odourleaa or haa a faint, aromatic odour and an intensely sweet taste, even in very dilute solutions.


Identification Teats

Water: Freely soluble ~thanol: Sparingly soluble

B. Diaaolve about 100 mg of the sample in 5 ml of 5i aodium ~droxide solution, evaporate to d.17neas and gently fuse the residue over a amall flame until it no longer evolves &111111onia. After the residue has cooled, dissolve it in 20 ml of water, neutralize the solution with diluted ~droohloric acid T.S. The addition of a drop of ferric chloride T.s. to the filtrate produces a violet colour.

59

Sod.iwa Sacch.arin (cont'd.) JECFA/51/11/1967

c. Nix 20 ag of th.e sample with. 40 ag of resorcinol, add. 10 drops of sulphuric acid. and. heat the aixture in a liquid. bath at 200° for 3 ainutes. After oooling, ad.cl. 10 •l of water and. an excess of sod.iwa ~d.roxid.e T.S. A fluorescent green liquid. results.

D. Dissolve o.5g of sample in 10 al water; add. l al dilute ~drochloric acid. T.S. and. allow to stand. l h.our. A whih crystalline precipitate is produced., which when washed. and. dried. for 4 hours at 105 aalts between 224° and. 230°.

E. Th• rHiciua obtained. b7 igniting a 2-graa sample givH positive tests for sodiva.

Water:

HaaY7 ••tale (as Pb)1

Seleniua1

12 to 16 percent b7 drying at 120°c tor 4 hours.

llot aore than 10 ag/q.

llot aore than 30 mg/q.

Weigh accuratal,y about 700 ag. of previousl;r dried. saapla and. transfer it quantitatively to a long-necked. flask, having a capacit,y of 200 al , and Mid 10 al of a 30 percent w/v solution of aodiua ~droxid.e in water; boil over a aall flaae for two llinutas, avoiding loss b7 evaporation; cool, ad.cl. 15 al of ~droohlorio acid., and. boil acaia for fift,y ainutea under a reflux condenser. Cool, rinse the condenser with 50 al of water, and. pass a current of air thrcuch the flaak to reaove acid. vapour. Connect with an &1111oaia distillation apparatus, add. 20 al of a 30 percent w/v solution of aodiua hydroxide in water, and. distil the amaoaia into 40 ml of 11/10 sulphuric acid; titrate the excess of acid with 11/10 aodiua ~droxici•, using solution of met~l red. aa indicator. Each •l of 11/10 sulphuric acid., neutralised b,y the aaaoaia in the distillate is equivalent to 20.52 •g of C7H411Jia03S•

60

METHODS OF AlfALYSIS

l. Acid. Value

Diaaolve about 10 graaa of the sample, accurately weighed, in 50 al of alcohol, previoualy neutralized to phenolphthalein with 0.1 B aod.ium hydroxide. Add l ml of phenolphthalein T.S. and titrate with 0.1 N sodium }IJ'droxide until the solution remains faintly pink after aha.king for 10 second.a, unless otherwise directed. Calculate the acid value (AV) by the formula:

AV : s,§1 x s w

in which S = the number of ml of O.l H sodium hydroxide conaumed. in the titration of the aample and W s the weight of the sample in graas.

2. Total Alcohola

Tranafer 10 graaa of a aolid sample, or 10 ml of a liquid aample, accurately weighed., into a 100 ml flask having a atand.ard taper neck. Add. 10 ml of acetic anhydride and l gram of anhydrous sodium acetate, mix these materials, attach a reflux condenser to the flask, and. reflux the mixture for l hour. Cool and add 50 ml of water at a temperature between 50° and 60° through the condenser, Shake intermittently during a period of 15 minutes, cool to room temperature, transfer the mixture completely to a separator, allow the layers to separate, and then remove and reject the lower, aqueous layer. Wash the essential oil laJ"•r successively with 50 ml of a saturate~ sodium chloride solution, 50 ml of a 10 percent sodium carbonate solution, and. 50 al of aaturated. sodium chloride solution. If the oil ia atill acid to moistened litmus paper, wash it with additional portions of aod.iwa chloride solution until it ia free from acid. Drain off the oil, dry it with anhydrous sodium sulfate, then filter it.

Weigh the quantity of acetylized oil specified in the monograph into a tared 125 ml Erlenmeyer flask, add 10 ml of neutral aloohol, 10 drops of phenolphthalein T.s., and 0.1 N alcoholic potassium hydroxide, dropwiae, until a pink endpoint ia obtained. If more than 0.35 ml is needed, reject the sample, and wash and test the remaining acetylized oil until its acid content is below this level. Prepare a blank using the same volwae of alcohol and indicator, and add l drop of O.l N alkali to produce a pink end-point, Measure 25.0 ml of 0.5 N alcoholic potassium hydroxide into each of the flasks, reflux them simultaneously for one hour, cool, and titrate the contents of eaoh flask with 0.5 N hydrochloric acid to the

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ciisappearance of the pink oolour. Calculate the percent of Tot al Alcohol s by the formula:

A = (b - Sl(lOO.) W - 21 b - S)

in which b = the number of ml of 0.5 H hydrochloric acid consumed in the titration of the blank, S: the number of ml of 0.5 H hydrochloric acid consumed in the titration of the sample, e = the equivalence factor given in the monograph, and W • the weight of the sample of the acetylized oil in. mg.

3. Deteminahon of Aldeb.Yd ..

BYdroXYlaaine BYdrochloride Solution. Dissolve 50 graas of hydro:Q<laaine hydroohloride, preferably recrystallized before uaing, in 90 ml of water and ciilute to 1000 ml with aldeh,yde-free alcohol. Adjust the aolution to a pH of 3.4 with 0.5 • alcoholic pota.siwa hydroxide.

Procedure. Transfer an aoourately weighed quantity of saaple specified in the monograph into a 125 ml ErleD111eyer flaak. Add 30 •l of HYdroXYl,-ipe BYdroohlo[ide Solution, •!x thorovghl.7, and allow to stand at room temperature for 10 mi nutes, unless otherwise specified. Perform a blank d.eteriaiDation si.llultaneoialy with the sample detel'llination. Titrat e the liberated hydrochloric aoid with 0.5 B alcoholic potaasiwa hydroxide to a greenish 7ellow end-point, iaing bro•ophenol blue T.S . aa the indicator, or titrate to a pH of 3.4 using a suitable pH mater. Calculate the percent of aldeh,yd.e (A) b7 the fol'lllula:

A • (S - b JClOOE) w

in which S • the number of ml of 0.5 B alcoholic potassiwa h,ydroxid.e consumed in the titration of the sample, b: the nWlber of •l of 0.5 H alcoholic potassium hydroxide consumed in the t itration of the blank, i • the equivalence factor given in the monograph, and W = the weight of the sample in mg.

4• Determination of ildeb.Yd .. and Ietones

4.1 H,ydro!,Jlaaine Method

Dissolve 20 grama of hydro:Q<lamine hydrochloride reagent grade or preferably freshly recr;yatall ised ) in 40 ml of water and ciilute to 400 ml with alcohol. Add, with stirring, 300 ml of 0.5 B alcoholic potassium hydroxide, and filter. Diaoard an,y- solution whioh i8 two da,ys or more old.

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Procedure. Transfer an accurately weighed. quantity of the sample specified. in the individual monograph into a 25c ml glass-stoppered. flask, add. 75.0 ml of the H.ydro;,ylamine Solution, and. stopper the flask. If the sample is an aldelU'de, allow the mixture to stand. at room temperature for l hour, unless otherwise directed in the monograph. If the sample is a~, attach the flask to a suitable condenser, and. reflux the mixture for l hour, unless otherwise directed in the monograph, and then cool to room temperature. Titrate the solution with 0.5 N hydrochloric acid to a greenish yellow end-point using bromophenol blue T.S. as indicator, or to a pH of 3.4 using a suitable pH meter. Perform a blank determination simultaneously with the sample determination. Calculate the percent of aldehyde or ketone by the formula:

AK (b - s)(lOOE) = w

in which AK= percent of aldehyde or ketone, b = the nU111ber of ml of 0.5 H hydrochloric aoid consumed in the titration of the blank, S = the number of al of 0.5 N hydrochloric aoid consumed in the titration of the sample, E = the equivalence factor given in the monograph, and W c the weight of the sample in mg.

4.2 Neutral Sulphite Method

Introduce 5 ml of substance into a 100 ml cassia flask fitted with a stopper. Ad.d. 50 al of a freshly prepared 40 percent sodium sulphite solution and 4 drops of phenolphthalein T.S. Heat in a boiling water bath, agitating frequently, for at least 2 hours. Neutralize the liberated. caustic soda froa time to time by adding 50 percent (by volume) acetic aoid solution, drop by drop, stoppering the flask to prevent lose of volatile material. Continue to heat until suoh time that any further addition of the sodium sulphite solution, plus 2 drops of phenolphthalein T.S. no longer causes pink colouration. Cool to room temperature. When the liquids have separated. completely, add. sufficient sodium sulphite solution to raise the lower level of the essential oil liberated into the graduated. neck of the flask. The quantity of essential oil in the neck of the flask, at room temperature, is used. to calculate the percentage of the aldehyde or ketone as follows:

% volume = °/,, weight =

100 - (a x 20)

100 - (a x 20) D d

in which a= the number of ml of separated oil in the graduated neck of the flask, D = density of aubstance combined. and. d: density of the essential oil.

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5. Determination of Eaters

5.1 Eater Determination

Transfer an accurately weighed quantity of the sample specified in the monograph into a 125 ml Erlenmeyer flaak containing a few boiling atones. Add to thia flask, and, simultaneously, to a similar flask for a blank teat, 25.0 ml of 0.5 H alcoholic potassium lqdroxide. Connect each flaak to a reflux condenser, and heat the mixtures on a steam bath for exactly 1 hour, unless otherwise directed in the monograph. Allow the mixtures to cool, add. 10 drops of phenolphthalein T.S. to each flask, and titrate the exoeea alkali in each flaak with 0.5 H }qdroohlorio acid. Calculate the percent of eater (E) in the sample by the formula:

E : (b - SJ(lOOe)

" in which b. the number of ml of 0.5 H lqdroohlorio acid oonauaed in the residual blank titration, S • the nuaber of ml of 0.5 I lqdroohlorio acid consumed in the titration of the sample, e • the equivalence faotor given in the monograph, and W = the weight of the aaaple in mg.

5.2 Eater Determination (High Boiling Solvent)

0.5 I Potassium HYclroxide f?olution. Dissolve about 35 graaa of potaaaiua lqdroxide in 75 al of water, add. 1000 ml of a suitable grade of monoetlql ether of diet}qlene glycol, and mix. -

Procedure. Transfer an accurately weighed quantity of the sample specified in the monograph into a 200 ml liirlenmeyer flask having a standard-taper joiat, and. add 2 glaaa beads and. 25.0 ml of 0.5 5 Pctaaaiwa HYCU'oltid.e Solution, allowing exaotl7 l minute tor drainage from the ~uret or pipet. Attach an air condenser, reflux gently for l hour, and. cool. Rinse down the condenser with water, using about 50 ml, then add phenolphthalein T.S. and titrate the excess alkali with 0.5 5 sulphuric acid to the disappearance of the pink colour. Perform a blank determination, using 25.0 ml of the alkali and 50 ml of water. Calculate the percent of eater in the sample by the formula (b - S)(lOOe)/V, in which b ia the number of ml of 0.5 5 sulphuric acid consumed. in the blank determination, Sia the nuaber of ml of 0.5 H sulphuric aoid. required. in the titration of the aaaple, e ia the equivalence factor stven in the monograph, and W ia the weight of the sample in mg.

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Saponification Value. Proceed aa directed for Ester Determination, Calculate the saponification value (SV) by the formula:

sv = (b - sl(28.05J w

in which band Sare as defined under Ester Determination, and W = weight of the sample in grams,

Eater Value. If the sample contains no free fatty acias, the aaponification value ana the ester value are identical, If a determination of the Acid Value (AV) is specified in the monograph, calculate the ester value (EVJ by the formula:

EV : SV - AV,

6. Determination of Linalool

Transfer a 10 ml sa111ple, previously dried with sodium sulphate, into a 125 ml glass-stoppered Erlenmeyer flask previoualy cooled in an ice bath, Add to the cooled oil 20 ml of dimethyl aniline (monomethyl-free) and mix thoroughly, To the mixture add 8 ml of aoetyl chloride and 5 ml of acetic anhydride, cool for aeveral minutes, permit it to stand at room temperature for another 30 ainut3s, then i111111erse the flask in a water bath maintained at 40° + l for 16 hours. Wash the acetylated oil with three 75 ml portions of ice water, followed by successive washes with 25 ml portions of 5 percent sulphuric acid, until the separated acid layer no longer becomes cloud,y or emits an odour of dimethyl aniline when made alkaline, After removal of the dimethyl aniline, wash the acetylated oil first with 10 ml of al in 10 solution of sodium carbonate and then with successive portions of water until the washings are neutral to litmus. Finally dry the acetylated oil with anhydrous sodium sulphate and proceed as directed for Eater Determination under 5,1, Calculate the percent of linalool (C1oH18o) by the formula:

1 7,707(b - S) = W - C.021(b - S)

in which L = percent of linalool, b = the number of ml of 0.5 N hydrochloric acid consumed in the residual blank titration, S: the number of ml of 0,5 N hydrochloric acid consumed in the titration of the sample, and W c the weight of the aa111ple in grams,

Note: When this method is applied to essential oils containing appreciable amounts of esters, perform an!!.!!£ Determination, 5.1, on a sample of the original oil and calculate the percent of total linalool by the formula:

1 7,707(b - S)(l - 0,0021E) = W - 0.021(b - S)

in which L • percent of linalool, E • the percent of esters, calculated as linalyl acetate (c12a2002J in the sample of the original oil, a.nd b, s, and Ware as defined in the preceding paragraph.

This entire procedure is applicable only to linalool and linalool-containing oils. It is not intended for the determination of other tertiary alcohols.

7. Determination of Phenols

Pipet 10 ml of the oil into a 100 al cassia flask, add 75 ml of potassiWD hydroxide T.S . and shake vigorously for 5 minutes to ensure complete extraction of the phenol by the alkali solution. Allow the mixture to stand for about 30 minutes, then add suffioient potassium hydroxide T.s. to raise the oily l~er into the graduated portion of the flask, stopper the flask , and allow it to stand overni ght. Read the volume of insolubl e oil to 0.05 ml. Calculate the percent, by volWDe, of phenol by the formula,

P : (10 - V) x 10

in which P = percent of phenols, by volume, and V • observed volume of insoluble oil in •l.

8. Chlorinated Compounda

Wind a 1.5 x 5 cm. strip of 20-aesh copper gauze around the edge of a copper wire. Heat the gauze in a non-lumi nous flame of a Bunaen burner until it glows without colouring the flame green. Perait the gauze to cool and re-ignite it several times unti l a good coat of oxide has formed. With a medicine dropper , apply 2 drops of the sample to the cooled gauze, ignite, and perait it to burn freely in the air . Again cool the gauze, add 2 more drops, and burn as before. Continue this process until a total of 6 drops has been added and ignited. Then bold the gauze in the outer edge of a Bunsen flame, adjusted to a height of 4 cm. Kot even a transient green colour i s i•parted to the flame.

9. Solubility in Alcohol

Transfer al.Cal sample into a calibrated l C ml glaaastoppered cylinder graduated in O.l •l aubdi visions, and add slowly, in .. all portions , alcohol of the concentration specified in the •onograph. Maintain the temperature at 20° and shake the cylinder thoroughly after each addition of alcohol. When a clear solution is first obtained, record the number of ml of alcohol required. Conti11ue the addition of the alcohol until a total of 10 ml has been added. If opalescence or cloudiness occurs during these subsequent additions of alcohol, record the number of ml. of alcohol at which the phenomenon occurs.

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10. Solidification Point

10.1 Scope

This method is designed to determine the solidification point of food graae chemicals having appreciable heats of fusion. It is applicable to chemicals having solidification points between -20° and +150°.

10.2 Definition

Solidification Point is an empirical constant defined as the temperature at which the liquid phase of a substance is in approximate equilibrium with a relatively small portion of the solid phase. It is measured by noting the maximum temperature reached during a controlled cooling cycle after the appearance of a solid phase.

Solidification point is distinguished from freezing point in that the latter term applies to the temperature of equilibrium between the solid and liquid state of pure compounds.

Some oh .. ical compounds have two teaperatures at which there may be a temperature equilibrium between the solid and liquid state depending upon the crystal form of the solid which is present.

stirrer

TT-150T mm.

2001100 1·_"'£

368 mm.

~~, ~cooling bath

air jacket

Fig. 1 - Apparatus for Determinir.g Solidification Point

,-length of

stirrer (as required)

4mm.

-T 3 loops

approx. 50 mm.

_l_

Fig. 2 - Stirrer

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10.3 Apparatus

The apparatus illustrated in Figure l consists of the following components.

Thermometer. A thermometer having a range not exceeding 30°, graduated in 0.1° divisions, and calibrated for 76 mm immersion. A thermometer should be so chosen that the solidification point is not obscured by the cork stopper of the sample container.

Sample Container. Standard glaas 25- x 150-mm teat tube with lip, fitted with a two-hole cork stopper to hold the thermometer in place and to allow •tirring with stirrer.

Air Jacket. A standard glaas 38- x 200,.mm test tube with lip, fitted with a cork or rubber stopper bored with a hole into which the sample container can easily be inserted up to the lip.

Cooling Bath. A 2-litre beaker or similar suitable container. Fill it with an appropriate cooling mediUIII such aa glycerin, mineral oil, water, water and ice, or alcohol-a.r;y ice.

Stirrer, The stirrer (Figure 2) consists of a 1-mm diameter {Band S gauge 18), corro•ion-resi•tant wire bent into a seriea of three loopa about 25 111111 apart. It should be made •o that it will move freely in the apace between the thermometer and the inner wall of the sample container, The shaft of the stirrer should be of a convenient length de•igned to pasa looaely through a hole in the cork holding the thermometer. Stirring ma,y be hand operated or mechanically activated at 20 to 30 •trokea per minute.

Aaaeably. Assemble the apparatus in such a wa.1 that the cooling bath can be heated or cooled to control the deaired temperature ranges, Clamp the air jacket ao that it is held rigidly just below the lip and i11111erae it in the cooling bath to a depth of 160 mm.

10.4 Preparation of Sample

The solidification point is usually determined on ohemioala as they are received. Some ma,y be hygroscopic, however, and require special drying.

Products which are normally aolid at room temperature must be carefully melted at a temperature about 10° above the expected aolidification point. Care should be observed to avoid heating in such a wa,y as to decompoae or distil any portion of a &ample,

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10.5 Procedure

Adjust the temperature of the cooling bath to about 5° below the expected solidification point. Fit the thermometer and stirrer with a oork stopper so that the thermometer is centred and the bulb is about 20 mm from the bottom of the sample container. Transfer a sufficient amount of the sample, previously melted if necessary, into the sample container to fill it to a depth of about 90 mm when in the molten state. Place the thermometer and stirrer in the sample container and adjust the thermometer so that the immersion line will be at the surfaoe of the liquid and the end of the bulb 20 .± 4 mm from the bottom of the sample container. When the temperature of the sample is about 5° above the expected solidification point, place the assembled sample tube in the air jacket.

Allow the sample to cool while stirring, at the rate of 20 to 30 strokes per minute, in such a manner that the stirrer does not touch the thermometer. Stir the sample cont-inuously during the r .. ainder of the test.

The temperature at firat will gradually fall, then become constant as cr.rstallisation starts and continues under equilibriwa conditions, and finally will start to drop again. Some chemicals m113 superoool slightly below (0.5°) the solidification point; as crystalliaation begins the temperature will rise and remain constant aa equilibrium conditions are eatabliahed. Other products m113 cool more than 0.5° and cauae deviation from the normal patte~ of temperature change. If the temperature rise exceed& 0.5 after the initial oryatalliaation begins, repeat the teat and aegd the melted compound with ... a11 cr.rstals of the sample at 0.5 intervals as the temperature approaches the expected solidification point. Crystals for seeding m113 be obtained by freezing a small sample in a teat tube directly in the cooling bath. It is preferable that seed of the stable phase be used from a previous determination.

Observe and record the temperature readings at regular intervals until the temperature rises from a minimum, due to supercooling, to a maximum and then finally dropa. The maximum temperature reading is the solidification point. Readings 10 second.a apart should be taken in order to establish that the temperature is at the maximum level and continues until the drop in temperature is established.

TEST SOLUTIONS

Barium Chloride T.S.

Diasolve 12 grama of barium chloride, BaCl 2.2H2o, in sufficient water to make 100 ml.

Bromophenol Blue T.s.

Diasolve 100 mg of bromophenol blue in 100 ml of dilute alcohol (l in 2) and filter if necessary. For pH determinations, dissolve 100 mg in 3.0 ml of 0.05 N sodium hydroxide, and dilute with carbon dioxide-free water to 200 ml.

Ferrio Chloride T.s.

Disaolve 9 grams of ferric chloride, Fec1 36H20, in sufficient water to make 100 ml.

HYdroohloric Acid T.s., Dil~ted

A solution containing 10 percent (w/v) of HCl. Prepare by diluting 236 al of hydrochloric acid (36 percent) with sufficient water to make 1000 al.

ll7drogen Peroxide T.S.

A solution containing between 2.5 and 3.5 grams of B202 in each 100 ml. It·~ contain auitable preservatives, totalling not aore than 0.05 percent.

Lead. Subaoetate T.s.

Triturate 14 gra.11s of lead oxide, PbO, to a smooth paste with 10 ml of water, and transfer the mixture to a bottle, using an additional 10 ml of water for rinsing. Dissolve 22 grams of lead acetate, Pb(C2H302)2.)H20, in 70 ml of water, and add the aolution to the lead oxide mixture. Shake it vigorously for 5 minutes, then set it aside, aha.king it frequently during 7 ciaya. Finally filter, and add enough recently boiled water through the filter to make 100 ml.

Phenolphthalein T.S.

Diasolve l graa of phenolphthalein i n 100 ml of alcohol.

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Potassium Hydroxide T.S.

Dissolve 6.5 grams of potassium ~droxide, KOH, in water to make 100 ml.

Sodium HYdroxi<i• T.S.

Dissolve 4.3 grams of soaium ~droxide, NaOH, in water to make 100 ml.

specifications and criteria for identity and purity of

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