PmmChment Lely 473.43 AOAC Official Method 990.28 Sulfites in Foods Optimized Monior-Witiams Method First Action 1990 Final Action 1998 (Applicable of determination of 210 ppm (ig) sulfites in Foods. Applicable in presence of other volatile sulfur compounds; not ap- plicable to died onions, looks, and cabbage.) Results ofthe interaboratory study supporting the acceptance ofthe ‘method Hominy, 9.17 ppm (ug) sulfites: 8,5 1.355 = 142; RSD, = 14.5% RSDy = 15.5% Fruit juice, 8.05 ppm ug) sui 4,5 1.3655 ~ 1.62; RSD, = 169%: RSD, =20.1% Protein (seafood), 1041 ppm (ug) suites: 5,7 LAT: 5, =2.77; RSD, = 14.1% RSDQ = 26.6% A. Principe “Method measures fie sulfite plus reproducible portion of bound sulfites, suchas carbonyl ation produits, in foods. Tes portion is nated With ofuxing HCI (a 1M) to conver sults to SO, Stteam (of Ny introduced below surface of refluxing solution sweeps SOs ‘rough water-cooled condenser and, vis bubbler attached fo con- denser, with 3% H,O, solution, where SO, is oxidized to H,SO,. Sulfite contents directly related to generated H,SO,, which deter. mined by titration with standardized NaOH solution. For verifea- tion, sulfate can be determined gravimetically as BaSO, 8. Apparatus (a) Disilation apparatus —{(Note: tn this method, back pres- sue inside aparat is limited to unavoidable pressured to height (f.3%H,0, solution above tip of bubbler (F) Keep back pressure as low as possible to avoid loss of SO; through leaks. Use thin film of| stopcock grease on sealing surfaces ofall ints except join between separatory funnel and flask. Clamp together each joint to ensure complete seal throughout analysis) Assemble apparatus (Figure 990.284), wiich includes (/) inlet adapter (A) with hose connector (Keates 183000), Aduptr provides means of applying head pres- sre above solution. Use of presure-equalizing dropping funnel is not recommended because condensate, perhaps containing SO: is deposited in funnel and side arm. (2) Separatory funnel (B), 210ml capacity. (3) Round -hotom fas (), {Ly withtho=24/40 tapered joints. (4) Gas inlet ube (D) (Kontes 179000) of suicient length o pennit introduction of Np within 25 em of botiom of Mask. (6) Allihn condenser (E) (Kontes 431000-2430), jacket length 300 mm. (6) Bubbler (F), fabricated from glass according to dimen- sons in Figure 990.28B. (7) Vessel (G),ca2.Semidand I8cm deep. (b) Buret—I0 mL (Kimble Gass, Ine, No, 17124-F) with over- flow tube and hose connections for Ascarte tube or equivalent air-scrubbing apparatus to permit maintenance of CO,-fee etme sphere over standardized 0.010M NaOH, (6) Chilled water circulaior—Chill condenser with coolant, such as methanol-water(20 +40, vv), maintained at <15°C. Cirew lating pump, Neslab Coolfow 33 (Neslab Instruments, Tn., PO Box 178, Portsmouth, NH 03801, USA), or equivalent, is suitable, ©. Reagents (a) Aqueous hycrochloricacid—4M. For each analysis, prepare 90 mL solution by adding 20 mL HCI to 60 mL deionized (18 meg- ‘ohm water (b) Methyl red indicator—Dissolve 250 mg methyl red in 100. etal (6) Standardized rant —0.010M NaOH. Certified reagent may bbe used (Fisher SO-5-284). Standardize solution with reference standard potassium acid phthalate, (@) Hydrogen peroxide soluion —3%, Fr each analysis, dlste 3 mL ACS reagent grade 30% HO, to 30 ml. with deionized (18megohm) water. Just prior to use, add3 drops methyl red indica- {orand titrate with 0.010M NaOH to yellow endpoint. end point is excooded,diseard solution. (¢) Mirogen—High purity, used with regulator maintain flow ‘of 200 iL mia. To guard against oxygen in Ny gas, use GC-1ype rap (Ony-Purge N [Alltech-Applied Science Laboratories, Inc}, or equivalent). ‘Alieratively, oxygen-serubbing solution, such as alkaline pyrogalol in gas- washing botle (Kimble Glas, Ie.) may be used. Prepare tapas follows (/) Add 4.5 g pyrogallel to trap. (2) Purge trap with N;for2-3 min. (3) Prepare KOH solution by adding 65 x Williams method: A, inlet adapter; B, separatory funnel; C, round-bottom flask; D, gas inlet tube; E, Alin con- denser; F, bubbler; G, vessel ‘5 2000 AOAC INTERNATIONAL[+2000 12s| Figure 990.288—Enlarged diagram of bubbler for Monier-Williams apparatus (lengths in mm). KOH 1085 mL H.0. (Caution: Heats generated) (4) Add KOH so- Tution trap while atmosphere of N; i malstained in ap. 2. Test Sample Preparation (a) Solids. Transfer 50 g food, or quantity that contains 500-1500 jg $0, to fod processor o Blender. Add 100 meth nol-water(S 95, vv) and briefly grind mixture. Continue grinding ‘or blending onl until fod is ehonped ito pisces small enough to ‘passthrough standard aper 24/40 joint of flak (C). (Wb) Liguids.—Min 50 g test portion, or quantity that contains 500-1500 ug SO; with 100 mL etbanol-water(5-+95, VW. (ote: Carry out test sample preparation and analysis as quickly ss possible to avoid loss of labile forms of sulfite) E. System Preparation Using apparatus assembled as shown in Figure 990.284, position ‘ask (C) in hsating mantle controled by power-epulating device (host) and add 400 ml. H0 to flask, Close stopcock of separs- tory funnel (B) and add 90 ml. 4M HCI a separatory funnel. Begin N, flow at 200.4 10 mL min. tite condenser coolant flow at this time. To vessel (G) add 30 mi 3% HyO,, which has been tivated 10 yellow end point with 0.010M NaOH. After 15 mia, apparatus and ‘water will be thoroughly deoxygenated and prepared tet portion tay be introduced into system F. Sample introduction and Distilation Remove separatory Funnel (B) and quantitatively transfer test por: tion in aqueous ethanol to ask (C)- Wipe tapered joint clean with laboratory tissue, quickly apply stopcock grease 10 outer joint of separatory fuel, and return separatory funnel to ask. Nitrogen flow through 3% #0, solution resumes as soonas separatory funnel {is reinseredinto appropriate joint in flask Examine cach joint to be sure that it ssl Use rubber bulb equipped with valveto apply head pressure above Clin separatory funnel. Open stopcoek in separatory funnel andet HCI low into ask. Continue to maintain sufficient pressure above acid solution to force solution into Mask, Stopeack maybe clase, if noeessary, fo purp up pressure ahove aid, and then opened again. ‘Close stopcock before last 2-3 mL drain out of separatory funn to _Buard against escape of SO, nto separatory funn! Apply power to heating mantle, Use power sting that causes 80-90 drops/min of condensate to returs fo Mask from condenser. ‘Let contents of flask boil 1.7 h, and then remove vessel (3). . Determination (2) Tiration.—Immediatlytieate contents of vessel (G) with (0.010M NaOH to yellow end point tha persist 220 s. Compute sul Fite content, expressed ing SOy fod (ppm), as follows: 0, ua (ppm) = 3209 =6=1000 ele wher 32.03 = miliguivalent weight of $0); Va=velume (a. of NaOH of molanty M requied to reach nd poi; 1000 ~ foro convert milegivalens to microcquvslets; weight =weight of tes porn noduced into tL ask (0) Gravimeric determination —Optional. Following ation, vise contents of vessel (0) iio 400m. beer. A drops IM HCIandoxces of itered 10% BaClslsion and let mixture and ovemight Wash rcipitate by desatation 3 ines wih bt water through weighed Gooch enc. Wash with 20 en. alcobol and 20 meee, and dry a 108-110°C sng BaSO, x274.46 ges portion 80>, me/e pm) = (6) Blank determination —Determne blank on ceagents bat by tation and gravimetrically, and correct results accordingly. H. Recovery Assays “To become familiar and proficient with method before routine us, analyze fod test portions containing known amounts of sulfite Perform analysis in manne tha precudes any los of sulfite by oxi- dation o reaction with components in food. Since sulfites are resc- tive with ai and food matrixes and lack stability, fori portions with table source of sult, not sodium sulfite or similar sals. So- dium hydroxymethylsulfoate (HMS), whic is bisulfite addition product of formaldehyde and is structurally similar to some com- bined forms of sulfite in foods, is wef or preparing stable fried test materials, Foranalsis, transfer 50g prepared test sample of sulfites food ‘0 Monicr-Williams ask. Add aliquot of agucous solution of HMS sodium salt. Analyze solution immediatly, LMS recoveries of 380% from food matrixes fortified at 10 pa/ are recommended to ensure accurate analytical data Reference: JAOAC 72, 470(1989) CAS-7446.09-5 (sulfur dioxide) (© 2000 AOAC INTERNATIONAL