"ire ee eer Orr Fon AOAC Cy, MeraLs ANo OrweR ELEMENTS —____* —— nn or etectrocless discharge lamp, (oy Halon cat = (©) Photomer:—Aspirate test and blank solutions, and record ‘A. Measure 4 of matrix standard solution containing 10 yg/mt. If this value differs from value of average of the six values used (0 calculate s by >2s, repeat measurement. If these values indicate drift, determine cause, correct it, and repeat calibration and tes blank readin {©} Caculations —Comect readings of test solution for blank Convert corrected 4 to pg/mL. trom calibration curve. Ov Concentration (mg/kg) Ni (or Cu) => inal volume test solution ag metal/mL. from curve, (50), and HW'= g test portion, References: JAOAC $3, 531(1970); $4, 658(1971) CAS-7440-50-8 (copper) CAS-7440-02-0 (nickel) 9.1.08 AOAC Official Method 999.10 Lead, Cadmium, Zinc, Copper, and Iron in Foods ‘Atomic Absorption Spectrophotometry ‘after Microwave Digestion First Action 1999 Final Action 2005 NMKL-AOAC Method [Applicable to determination of Zn, Cu, and Fe in a variety ‘of foods by mierowave digestion and flame atomic absorption spectrophotometry (FAAS), and Ca and Pb by microwave digestion and graphite furnace atomic absorption spectroscopy (GFAAS). “Method is capable of determining these elements at concentrations above approximately Pb (0.4), Cd (0.01), Zn (4), Cu (3), and Fe (7) mgkg. Method is not applicable to foods with a fat content 240%. Not applicable to milk powder] ‘See Table 999.10A for results of the interlaboratory study supporting acceptance of the method. Caution: Digestion vessels must cool for an appropriate time before opening in order to avoid burns from hot and ‘corrosive vapors. Always gently add acid to water. Maintain safe distance from furnaces equipped with ‘Zeeman background correction when the magnet is ‘on. Consult manufacturer's instructions to determine safe distance, which varies for different instruments, See Appendix B, Laboratory Safety, for safe use of compressed gases, inorganic acids, and atomic absorption spectrometer. A. Principle Products are digested with HNO, and H,O, under pressure in a closed vessel heated by microwaves. Solution is diluted with HO. Pb and Ci are determined by GFAAS. Zn, Cu, and Fe are determined by FAAS. B. Apparatus (8) Atomic absorption spectrophotometer—With sir-acetylene bumer oF nitrous oxide-acetylene bumer for flame (FAAS, soe Table 958.108) ands gph Furace fo electrothermal (FAAS, see Table 999.10C) determinations, with appropriate ‘ (oonatomie) correction ace ‘DOT AOAC INTERNATIONAL © sen -—Designed for laboratory yy eh athews, NC. USA) Cheek Tr ee ifthe measured ene ion adjust the ProBram: Filla laggy Hath 1-000 KE Water oom eg U7) Place beaker in microwayeet = for 2 min. Take beaker out of yc"! ature (7,)- The delivered power 2 way heat water i water, and measure tem po3sx(T,-T) (a Tefon digestion vessels 100 mL, withstanding pag, of atleast 1.4 MPa. ey olumetric flasks —25 antl 1000 mL. ass or plastic. () Funmels.— Ba ih (a Plt ots Polystyrene botles with igh fg, lids, 30-100 ml or equipment for freeze-drying with HINO, or HCl, to avoid metal contamination. . Reagents Reagents should be of at least analytical reagent arate (a) 1 equivalent. rferably ultrapure (suprapur), oF equi Pia) Water Redisilled or deionized, =18 MQ-em. (b) Nitric acid 65% (W/W). {@) Nitric acid —0.1 M. Dilute 7 mL concentrated HNO, with water to TL (@) Nitric acid —3 M. Dilute 200 mL. concentrated HNO, 0, with water to 1 L. (© Hydrogen peroxide —30% (w/w). (9 Zine standard solution—1 mg/mL. Dissolve 1.000 g Zi 14 mL water + 7 mL nitric acid, (b), in 1 L volumetric flask. Dilue to volume with water, [Nofe: Commercially available standan solutions for AAS (e.g., BDH Chemicals Ltd, Poole, UK) may be used for all metal standard solutions.) (g) Copper standard solution —1 mg/mL. Dissolve 1.000 0 in 7 mL nitric acid, (b), in 1 L volumetric flask. Dilute to volume with water. (h) Iron standard solution—1 mg/mL. Dissolve 1.000 g Fe 14 mL water + 7 mL nitric acid, (), in 1 L volumetric flask. Dit to volume with water (@ Lead standard solution —1 mg/mL. Dissolve 1.000 8 P°#* 7mLHNO, (®) in volumetric tak and dilute to volume ™# co) Cadmium standard solution —1 mg/mL. Dissolve 1.001% fea mL Water +7 mL HNO,, (b), in IL volumetric i or volume with water.” ) Working standard solutions.—(1) For ysis Dive standard solutio -flameanalrsis standard, (Qj), with 0.1 M HNO, (@), to a range ‘of standards rohean of the element to be determined. 21° OM tegcitce anayss.—Ditut standard solutions. (0-0 range ofthe i 8 ange oF standards thar covers HE lement to be determi D. Procedures letermined (8) Cleaning py wr solution: Soy gf wceaere-—(1) For glass an plastion mL. concentrated HNO,, C(b), + 4500 mL deg water, * (a). Wash first with water and detergent. Rins® "!ye 95 OF ANALYSIS (2010) Meta AnD OTHER ELEMENTS: yore ee _- Chaptor 9.9.17 _ result for lod, cad Ta ooo {or oad, cadmium, zine, copper, and iron in foods ee ‘annttong Moan, mong mt Owen 8, mm ROD, es eee ee ihe RR Horkat oa Een | 008 aa 18S MS oso is. 0.088 0001 575902506281 ‘ ae oan © 00630008 «16 «25 oer 1.87 ine : 0.086 2 ow 124 ee oa 120 0.13 a 036 192 | stom 1620120 026 16 ors 108 | prio 0° 20.01 oo 12 «1 0.0034 28 0.0097 0.91 1 gO vee tes 1300025034 15 «= 200.070 0.084 0.96 aoe O17! 11-2 00078 0022 46 «13 Omae 0063088 | oa 21 12 0.035 7 0.099 0.85 | poor o42 2 0.053, " 0149 062 ae o7e4 1210050 0.105 65 14 «(0.140.204 085 sh za 450 120 oat e4 14 0st 2) a powder 33 140 33 93 e1 1.00 | iets? oaien io oe ao eeemeee etl le | ustroom 9 40 30 53 a4 06t heat bran Tag PMS 1 ee Se She. ABs Wear PD 4 OF. ovine msde 473. Mi. 8 25 7 70 023 | ie tig 2228 BH AB 7H 5 8B an 202 oz 40 0.094 9 02 1.98 ovine muscle 209 «6 «0 oar 64 047047 wat bran Fe 4 ean.ed octOBtdlew A2icokT.OMir BB ut 2815 0-70 Nustroom a7 4H 22 57 60 062 ets? eduowantial 28 0g8S 18 a ee lat iver Gs TE 0. BB MP rte. 8B ult PAcne ved, «O88 fs) Fish 7 haeten ©. 13 7 a5 144 Bovine muscle 70 «12,:—«O 8A " 23192 Mushroom 4055 1 OO 79 15 2 © 095 Wheat ran GAY MB E88 ye Bn, BAe Bb es Ut BB OS Diets? Ser or zi M8 bie 4.0 sd oir BB Snr Ds or 8S ver peers War meee ore o2 Tasca GAs SRST Vas RSD, ote ava Glee FSR vel deteranaons Siac Bans stu eset die water, then with acid solution. Finally ine a éeionized water. (2) For Teflon digestion Se i ones wash, with deionized water, keep eng i 0.1 M HNO,, C(o), for atleast 30 min, rinse eae yaa It ese dy nen oe fie application, depending on the Ntheiyemi towers the same digestion vessels Se tee mona products, e.g, sludge it ay be N severe cleaning procedure, e.g. heating wih Sy ri stesed HNO, C(b), The instrument 4 tailed instructions for such cleaning ean ‘Nanaet product iso be analyzed fresh, proceed to "" Otherwise, continue at (e), Drying. (6 Drving Dry to constant weight in drying oven at 105°C, oF frecze-y, Freezedrying is usually preferable because it renders the product less compact an easier to homogenize, If final result iz based on fresh weight, weigh test portion before and after drying to obtain water content: 4,0- 4.100 Ww y= water content ofthe test portion (%6); = weight rahe tet portion (a): H= weiuht after drying (@)- (@ Homogenizaion—Homogenize products’ using onoontaminating equipment. Check for leached metas if the consists of metal parts. where H,0, apparatus ‘©2016 AOAC INTERNATIONAL(METALS AND OTHER ELEMENTS Chapter 9, p. 18 Table 999.108. Instrumental parameters for FAAS. ee Metal Wavelength, rm a 738 cu 3247 Fe Air-acetylene, oxidizing 248.3 Fe N,0-acetylene, oxidizing 248.3 (©) Digestion —Weigh 02-0.5 g dry material into digestion vessel. If water-containing materials ae used, maximum weight is restricted to 2g, but dry matter content must never exceed 0.5 g. For ‘example, if product has a water content of 50%, take a maximum of 1 g(=0.5 g dry matter). Ifa product has a water content of 95%, take 2 g (<0.5g dry mater). When unknown products are digested, too much solids may cause the safety membrane in the digestion vessel to rupture, ‘Add S mL HNO,,C(b), and 2 ml 30% H,0,, C(e). Close vessels, place vessels in holder, place vessel holder in microwave oven, and close door Set oven program according tothe parameters given in ‘Table 99.10D and start program. ‘The program is valid only when 12 vessels are being digested simultaneously. If fewer ae being digested, the remaining vessels ‘must be filled with reagent blank. When a microwave oven other ‘than the one given as an example is used, it may be necessary to use 4 slightly different time/power program. Remove digestion vessels from microwave oven and let cool thoroughly before opening them. Open vessel and rinse down lid and walls into container. Transfer solution to 25 ml. volumetic flask and dilute to mark with deionized water. Then, transfer solution to plastic container. Treat blanks inthe same way as tests. ‘One blank shouldbe included in every set, (©) Ditution.—If est solution needs to be further diluted (due to high metal concentrations), dilute with 3 M HNO, C(d), in order to ‘maintain same acid concentration prior to metal determination, (g) High acid concentration is environmentally undesirable and may depress the analytical signal. Reduce acid strength by diluting the test solution 1/2 with 0.1 M nitric acid and standard solutions 1/2 with 3_M nitric acid. The tests and standards are thereby brought to the same acid concentration. Matching of acid concentrations is important when a calibration curve is used, (®) Atomic absorption spectrophotometry:—Use of flame ot ‘graphite furnace technique is determined by the concentration of the metal to be determined. Flame technique should be used as far a possible, since this technique is less sensitive to interference than the GFAAS. The most appropriate wavelength, gas mixture temperature program, and other instrumental parameters for each ‘etal are found in the manual provided with the instrument, ‘Always use background correction, ‘Table 999.106. Instrumental parameters for GFAAS Temp. (Cy ramphold() ‘sting —Aomizaton Ch Metal Wavelength, “step step 0) sae Fan $s — ca 2288 ——-350/15-10 120004 2509 ——_— . $e rr o0 (©2016 AOAC INTERNATIONAL AOAC OFFICIAL METHODS OF ANALY % a emer et Sa in can of sana rmmeants, of which at Teast (WO are 5 of at est te Pest standard should be 3s cow et sl The ve concentration * ion approximately half of the highes, Tune eee Pe mes of Sanda aha A simpli tched standard curve, which is applicable yy 4 2 naa od andr ston ge he andl atone Ths cae en ferred wo origin and is used as the standard curye fy gy Sl fia followed and that have been Te sone! Propon ‘The matrix-matched standard curve and the test solutions, Te ma ee concetton O8 Mos oer nto gue Tae eomcetaen Of 2a, are usually at levels suitable for determination by Faas. Whe calibration curve is to be used, standards and test solutions my ee oe nid concentration. Ne a arene acted by Hees fan matrix, use either the method of standard addition or ma, a ee. When experiencing toree Irises “oxidizing nitrous oxide acetylene flame may be an alternative, ce oem The eile required for determination of Pb and Cd in foods. Use pyrolytic, etc wit satura, Stace tbe metod rede san ip ana of th analy it snay fequeny be cee at trabaciminon of eg, Ca The method of andar sdane Secor a, an enim GOES Ctoarasame caltenon ves of pure woking suicartll mbes otes cave of trict poder). cannons tt be ote wi Tinea range when the method of addon wed Program the autosampler to deliver a volume that gives a lage an absorance as posible within the Tuer ray producing a background absorbance not larger than approxima) O15 absatance ints. Multiple injection may eee absorbance a very low concentrations, Evalue ech nv i by means of ash- and atomization-curves in order to optimize the Dili Gani ara E, Calculations and Evaluation of Results Will ys sume, Caleulate the concentration (C) of metal in the tet om according to the formula where C = concentration is 4 fon in the test sample (mets: & Free tation i the test solutions (mg/L); d= dilation than Concentration in the blank solutions (mg/L); m= weit the teu penton lank solutions (mg/L); ; 250, 2 3 630 f 500 2 4} JcjAt METHODS OF ANALYSIS (2016) por pee nt Sc Di ten a crave ano Ornen Bu ‘Chapter 1)» Reference: J, ADAC Ii BD, 11892000) ei tion of the limit of detection in the test tution has been diluted, the dilution factor (ap has 94.08 fe the et potion was died and he rca AOAC Offical Method 000.49 pen i a weigh conect socording 6 the followin Lead, Cadmium, Copper, Iron, and Zinc sue eon sh wei oor - in Foods eu 10-10% Atomic Absorption Bpectrophotometry i too After Dry Ashing «= camestato In the tat potion carectd to feah Fret Action 1000 oe gy onthe watt ote et pan) Fina! Action 2008 fant iia of Os fool tes ca fg. ete is a ual Mink ootctoninn ec amie ge woer of la mua be analyend bare DL Fe cacarapigilbett ener ecy Woe NMKL-AOAC Method ination of Zn, Cu, and Fe in a variety of foods by dry ashing and flame atomic absorption spectrometry (FAAS), and Cd and Pb by dry ashing and graphite furnace ‘atomic Absorption spectrometry (GFAAS). Method is capable of ‘determining these elements ut concentrations above approximately [Applicable to de Ph (0.3), Cal (0.1), Zn (1), Cu (5), and Fe (4) mg/kg.) wie SH.11A._Interlaboratory study results fork cadmium, copper, Iron, and zinc In foods to Anaiye Mean, fe Sample ronge mgkg =a? REDS RSD, —_ORF_—_HorRat ®EGERIS__Wer pasleimiik powder S004 “0040-10018 001946 48 0.052 0.082 1.70 ‘Apple sauce: Shai 10 0.10 38 oz 197 Minced fish 05310 ont 20 ost 114 Wheat bran om 2 0.056 60 018 226 Simulated diets DFE 0.248 © 10(1] 0034 0048 14 = 200096014 1.02 corAAS Liver paste 2005 ott 0.0088 2 0016 0.48 Minced fishwheat bran 0178 lt] 0021 0032 12 «18 «0.058.089 O87 Simulated diets DFE obip 6 “aio™ (ts ees a7 oat ad BAFMS —Lwerpasteminced ish 20.7 663 13035080 63780884 Apple sauce 0699 1113) 0.087 68 013 oat Wheat bran m5 12 49 68 14 ost Mik powder 3501242] 28 80 79 0.86 Simulated diets OVE gra] 088 13118 3881987 oe CHFMAS Apple saucelminced fish 202 0240 © 12I1) 0.078 0076 32 wk tozt 1.63 Mik powder ost 142] 023 45 085 286 Liver paste 5361312) 037 69 10 086 Wheat bran 95212 0.98 10 27 0.88 veggies OF 44 12] 16 434 36 «69 48a? ver paste Ey ms 4 28 " 78 112 Minced fishimik powder 399 118] 046 054 241.08 Wheat bran 1241911] 4 " 39143 = Simulated diets DIE gible neler cece ie ees "ange SSSRrRS ring ar aran eeu Das "ay standard deviation ; ana oviaion Rete Le “hay oh "Speatabity standard deviation, Sereducbty standard deviation ee (© 2016 AOAC INTERNATIONAL —