47.3.43 C.
Reagents
AOAC Official Method 990.28 (a) Aqueous hydrochloric acid.—4M. For each analysis, prepare
Sulfites in Foods 90 mL solution by adding 30 mL HCl to 60 mL deionized (18 meg-
Optimized Monier–Williams Method ohm) water.
First Action 1990 (b) Methyl red indicator.—Dissolve 250 mg methyl red in
Final Action 1994 100 mL ethanol.
(c) Standardized titrant.—0.010M NaOH. Certified reagent may
be used (Fisher SO-5-284). Standardize solution with reference
(Applicable of determination of ≥10 ppm (µg/g) sulfites in foods.
standard potassium acid phthalate.
Applicable in presence of other volatile sulfur compounds; not ap-
(d) Hydrogen peroxide solution.—3%. For each analysis, dilute
plicable to dried onions, leeks, and cabbage.)
3 mL ACS reagent grade 30% H2O2 to 30 mL with deionized
(18 megohm) water. Just prior to use, add 3 drops methyl red indica-
Results of the interlaboratory study supporting the acceptance of the tor and titrate with 0.010M NaOH to yellow end point. If end point is
method: exceeded, discard solution.
Hominy, 9.17 ppm (µg/g) sulfites: (e) Nitrogen.—High purity, used with regulator to maintain flow
sr = 1.33; sR = 1.42; RSDr = 14.5%; RSDR = 15.5% of 200 mL/min. To guard against oxygen in N2 gas, use GC-type trap
(Oxy-Purge N [Alltech-Applied Science Laboratories, Inc.], or
Fruit juice, 8.05 ppm (µg/g) sulfites:
equivalent).
sr = 1.36; sR = 1.62; RSDr = 16.9%; RSDR = 20.1%
Alternatively, oxygen-scrubbing solution, such as alkaline
Protein (seafood), 10.41 ppm (µg/g) sulfites:
pyrogallol, in gas-washing bottle (Kimble Glass, Inc.) may be used.
sr = 1.47; sR = 2.77; RSDr = 14.1%; RSDR = 26.6%
Prepare trap as follows: (1) Add 4.5 g pyrogallol to trap. (2) Purge
trap with N2 for 2–3 min. (3) Prepare KOH solution by adding 65 g
A. Principle
Method measures free sulfite plus reproducible portion of bound
sulfites, such as carbonyl addition products, in foods. Test portion is
heated with refluxing HCl (ca 1M) to convert sulfite to SO2. Stream
of N2 introduced below surface of refluxing solution sweeps SO2
through water-cooled condenser and, via bubbler attached to con-
denser, with 3% H2O2 solution, where SO2 is oxidized to H2SO4.
Sulfite content is directly related to generated H2SO4, which is deter-
mined by titration with standardized NaOH solution. For verifica-
tion, sulfate can be determined gravimetrically as BaSO4.
B. Apparatus
(a) Distillation apparatus.—(Note: In this method, back pres-
sure inside apparatus is limited to unavoidable pressure due to height
of 3% H2O2 solution above tip of bubbler (F). Keep back pressure as
low as possible to avoid loss of SO2 through leaks. Use thin film of
stopcock grease on sealing surfaces of all joints except joint between
separatory funnel and flask. Clamp together each joint to ensure
complete seal throughout analysis.) Assemble apparatus (Figure
990.28A), which includes (1) inlet adapter (A) with hose connector
(Kontes 183000). Adapter provides means of applying head pres-
sure above solution. Use of pressure-equalizing dropping funnel is
not recommended because condensate, perhaps containing SO2, is
deposited in funnel and side arm. (2) Separatory funnel (B),
≥100 mL capacity. (3) Round-bottom flask (C), 1 L, with three 24/40
tapered joints. (4) Gas inlet tube (D) (Kontes 179000) of sufficient
length to permit introduction of N2 within 2.5 cm of bottom of flask.
(5) Allihn condenser (E) (Kontes 431000-2430), jacket length
300 mm. (6) Bubbler (F), fabricated from glass according to dimen-
sions in Figure 990.28B. (7) Vessel (G), ca 2.5 cm id and 18 cm deep.
(b) Buret.—10 mL (Kimble Glass, Inc., No. 17124-F) with over-
flow tube and hose connections for Ascarite tube or equivalent
air-scrubbing apparatus to permit maintenance of CO2-free atmo-
sphere over standardized 0.010M NaOH.
(c) Chilled water circulator.—Chill condenser with coolant, Figure 990.28A—Apparatus for optimized Monier-
such as methanol-water (20 + 40, v/v), maintained at ≤15°C. Circu- Williams method: A, inlet adapter; B, separatory funnel;
lating pump, Neslab Coolflow 33 (Neslab Instruments, Inc., PO Box C, round-bottom flask; D, gas inlet tube; E, Allihn con-
1178, Portsmouth, NH 03801, USA), or equivalent, is suitable. denser; F, bubbler; G, vessel.
© 2000 AOAC INTERNATIONAL
Use rubber bulb equipped with valve to apply head pressure above
HCl in separatory funnel. Open stopcock in separatory funnel and let
HCl flow into flask. Continue to maintain sufficient pressure above
acid solution to force solution into flask. Stopcock may be closed, if
necessary, to pump up pressure above acid, and then opened again.
Close stopcock before last 2–3 mL drain out of separatory funnel to
guard against escape of SO2 into separatory funnel.
Apply power to heating mantle. Use power setting that causes
80–90 drops/min of condensate to return to flask from condenser.
Let contents of flask boil 1.7 h, and then remove vessel (G).
G. Determination
(a) Titration.—Immediately titrate contents of vessel (G) with
0.010M NaOH to yellow end point that persists ≥20 s. Compute sul-
fite content, expressed in µg SO2/g food (ppm), as follows:
32.03 × VB × M × 1000
SO2, µg/g (ppm ) =
weight
Figure 990.28B—Enlarged diagram of bubbler for
where 32.03 = milliequivalent weight of SO2; VB = volume (mL) of
Monier-Williams apparatus (lengths in mm).
NaOH of molarity M required to reach end point; 1000 = factor to
convert milliequivalents to microequivalents; weight = weight, g, of
KOH to 85 mL H2O. (Caution: Heat is generated.) (4) Add KOH so- test portion introduced into 1 L flask.
lution to trap while atmosphere of N2 is maintained in trap. (b) Gravimetric determination.—Optional. Following titration,
rinse contents of vessel (G) into 400 mL beaker. Add 4 drops 1M
D. Test Sample Preparation
HCl and excess of filtered 10% BaCl2 solution, and let mixture stand
(a) Solids.—Transfer 50 g food, or quantity that contains overnight. Wash precipitate by decantation 3 times with hot water
500–1500 µg SO2, to food processor or blender. Add 100 mL etha- through weighed Gooch crucible. Wash with 20 mL alcohol and
nol–water (5 + 95, v/v) and briefly grind mixture. Continue grinding 20 mL ether, and dry at 105–110°C.
or blending only until food is chopped into pieces small enough to
pass through standard taper 24/40 joint of flask (C). mg BaSO 4 × 274.46
SO2, µg/g (ppm) =
(b) Liquids.—Mix 50 g test portion, or quantity that contains g test portion
500–1500 µg SO2 with 100 mL ethanol-water (5 + 95, v/v).
(Note: Carry out test sample preparation and analysis as quickly (c) Blank determination.—Determine blank on reagents both by
as possible to avoid loss of labile forms of sulfite.) titration and gravimetrically, and correct results accordingly.
E. System Preparation H. Recovery Assays
Using apparatus assembled as shown in Figure 990.28A, position To become familiar and proficient with method before routine
flask (C) in heating mantle controlled by power-regulating device use, analyze food test portions containing known amounts of sulfite.
(rheostat), and add 400 mL H2O to flask. Close stopcock of separa- Perform analysis in manner that precludes any loss of sulfite by oxi-
tory funnel (B) and add 90 mL 4M HCl to separatory funnel. Begin dation or reaction with components in food. Since sulfites are reac-
N2 flow at 200 ± 10 mL/min. Initiate condenser coolant flow at this tive with air and food matrixes and lack stability, fortify portions
time. To vessel (G) add 30 mL 3% H2O2, which has been titrated to with stable source of sulfite, not sodium sulfite or similar salts. So-
yellow end point with 0.010M NaOH. After 15 min, apparatus and dium hydroxymethylsulfonate (HMS), which is bisulfite addition
water will be thoroughly deoxygenated and prepared test portion product of formaldehyde and is structurally similar to some com-
may be introduced into system. bined forms of sulfite in foods, is useful for preparing stable fortified
F. Sample Introduction and Distillation
test materials.
For analysis, transfer 50 g prepared test sample of sulfite-free food
Remove separatory funnel (B) and quantitatively transfer test por-
to Monier-Williams flask. Add aliquot of aqueous solution of HMS
tion in aqueous ethanol to flask (C). Wipe tapered joint clean with
sodium salt. Analyze solution immediately.
laboratory tissue, quickly apply stopcock grease to outer joint of
HMS recoveries of ≥80% from food matrixes fortified at 10 µg/g
separatory funnel, and return separatory funnel to flask. Nitrogen
are recommended to ensure accurate analytical data.
flow through 3% H2O2 solution resumes as soon as separatory funnel
is reinserted into appropriate joint in flask. Examine each joint to be Reference: JAOAC 72, 470(1989).
sure that it is sealed. CAS-7446-09-5 (sulfur dioxide)
© 2000 AOAC INTERNATIONAL
