47.6.
12
AOAC Official Method 941.10
Saccharin in Food
Qualitative Tests
First Action 1941
Final Action
A. Organoleptic Test
Acidify with HCl 50 mL nonalcoholic liquid food or aqueous ex-
tract of 50 g solid or semisolid product, 973.29A(c) (see 47.6.13),
and extract with three 25 mL portions ether. Wash combined ether
extracts once with 5 mL H2O, transfer to small beaker or evaporating
dish, let ether evaporate spontaneously, and taste residue. (Presence
of 20 mg saccharin/L or kg of original test sample can usually be de-
tected by its sweet taste.) Confirm by heating with NaOH and detect-
ing salicylic acid formed thereby as in B.
B. By Conversion to Salicylic Acid
Acidify with HCl 50 mL nonalcoholic liquid food, or equivalent
volume aqueous extract, 973.29A (see 47.6.13), and extract with
3 portions ether as in A. Dissolve residue remaining after evapora-
tion of ether in little hot H2O and test small portion of solution for
salicylic acid as in 975.30A or B (see 47.3.34).
Dilute remainder of solution to ca 10 mL and add 2 mL H2SO4
(1 + 3). Heat to boiling and add slight excess of 5% KMnO4 solution
dropwise; partly cool solution, dissolve ca 1 g NaOH in it, and filter
mixture into Ag dish (Ag crucible lids are suitable). Evaporate to
dryness and heat 20 min at 210–215°C. Dissolve residue in H2O,
acidify with HCl, and test ether extract for salicylic acid as in
975.30A or B (see 47.3.34). By this method all so-called “false sac-
charin” [J. Am. Chem. Soc. 26, 1627(1904)] and any salicylic acid
naturally present (also added salicylic acid when not present in too
large amount) are destroyed, whereas 5 mg saccharin/L is detected
with certainty.
C. Phenol-Sulfuric Acid Test
(Applicable to nonalcoholic beverages, semisolid preparations,
and baked goods.)
Prepare ether extract of test portion as follows:
(a) Nonalcoholic beverages.—Add 3 mL HCl to 25 mL test
portion in separator. If vanillin is present, remove by extracting
with several portions petroleum ether. Discard petroleum ether.
Extract with 50, 25, and 25 mL ether-petroleum ether (1 + 1).
Wash combined ether extracts once with 5 mL H2O, remove ma-
jor portion of solvent, transfer to 30 mL beaker, and evaporate at
room temperature.
(b) Semisolid preparations.—Transfer 25 g test portion to
100 mL volumetric flask with small amount hot H2O and add enough
boiling H2O to make ca 75 mL. Let mixture stand 1 h shaking occa-
sionally. Then add 3 mL CH3COOH, mix thoroughly, add slight ex-
cess (5 mL) of 20% neutral Pb(CH3COO)2 solution, dilute to volume
with cold H2O, mix, let stand 20 min, and filter. Transfer ≥60 mL fil-
trate to separator and proceed as in (a).
(c) Baked goods.—Grind 25 g test portion, mix thoroughly with
50 g washed and ignited sea sand, and extract with petroleum ether in
Soxhlet apparatus until essentially fat-free (1–2 h). Transfer ex-
tracted mass to 300 mL Erlenmeyer, add 100 mL alcohol, and reflux
on boiling H2O bath 30 min, shaking frequently. Filter through
Büchner containing 7 cm Whatman No. 2 paper wet with alcohol.
Transfer alcohol filtrate to 100 mL beaker, evaporate to 12 volume,
add 50 mL H2O and enough 10% Na2CO3 solution to make alkaline,
and evaporate to 50 mL. Transfer aqueous solution to separator and
proceed as in (a).
To residue remaining after evaporation of solvent add 5 mL phe-
nol-H2SO4 reagent (pure colorless crystalline phenol dissolved in
equal weight H2SO4) and heat 2 h at 135–140°C. Cool, dissolve in
small amount of hot H2O, and pour into ca 250 mL H2O. Add small
amount of Filter-Cel, let stand 3 h or overnight, and filter. Make al-
kaline with 10% NaOH solution and dilute to 500 mL. Magenta or
reddish-purple color develops if saccharin is present. Yellow, buff,
or pale salmon shade is not significant.
References: Z. Nahr. Genussm. 31, 67(1915).
JAOAC 24, 326(1941).
CAS-81-07-2 (saccharin)
© 2000 AOAC INTERNATIONAL