auirs ano FRUIT PRODUCTS Chapter 37,9. 14 37.41.46 AOAG Official Method 986.13, Quinie, Malic, and Citric Acids in Cranbériy Juice Cocktail and Apple Juice Liquid Chromatographie Method rst Action 1286, Final Aetion 1989 A. Principle Juice is eluted through disposable cartsidge to remove interferons, Sted, and injected into guid chromatographic system, Quinio, alle, and cite acids are separated by using 2 Feversed-phase LC columarin tandem with UV detection at 214 nm. ‘Conpouadsare quantitated by compasson withexternal standards 18. Apparatus and Reagents (a) Liguid ciemezograph—Sysem expipped with Model 77251 inject, Medel 2487 vale wavelength decor operable at 24 nm, 0.1 AUFS (Waters Acceciates, Ino) and computing integrator (Glowiet- Packard Integrator 3390 (replaced by No. 3396B], of enilea. (b) Anabtica! columns—{2) Sopeloosil LC-18, or equivalent, 5 ym particle si, 28cm 4.5 nm, i tandem with and fellowed by (G) RadiabPak CUE caroiye CWuers Associates, ne), 5pm parle som 10 em lon, sed with Radial Compression Module, Raa-Pak CCiseartidge oa be substied by any standard 25 0°30 cmstaniss see evered;phave CUS cotumn wits 10 um patil size. Connect BioRod reverse phase misroguted column (ODS-10) ahead of column. Mabile pase: phosphate buffer a 0.8 mini, sensitivity 01 AUES, (© Disposable ctvrtdges —Sep Pak C18 (Waters Associate, ne (@) Chemicolr—Porasium phosphate monobasic (KH,PO, 85% #3P0, muethanal, CH,CN, Filter ll solvents through aqueoas cor organi OAS yu iter (9 LCmabile phase.—Phosphate ber, 2M KH, PO. p24 ‘Weigh 272 g KH;POq in beaker Add HO to 950 aL. Using pt neler nd H;PO,, adjust to pH 2.4. Pour ito UL graduate andadjst to votume wit HO; ier (. Preparation of Test Solution aid Standarts (a) Working standard solutions. —Accurtely weigh, to neaest 0.1 mg, 0.200 g ACS grade quite, malic, and citi acids. Ditate combined avis to 100 ral ia volumetric ask with 80; files. (b) Test soluttont—Conttion carsidge by eluting 10 ml. CHGN-H,0 (50 + 50 trough 10 mL Luer-Lok syringe, Remove sen and pass 10 mi. sir rough cartridge. Blue 10 mL test portion though conditioned carrdge. Diseara Sst 45 mb and follect next 4-$ mE_ Filter tet elution for LC znlyss 0. Determination CCeodition syst with 100% methanol (or methano}-H0 [70+ 30) followed by 1,0 and then phosphate buffer Reverse order at ‘endof working é2y; never lt meth! aloakol come into contact with (phosphate bufer Operating conditions: Dow rate 0.80 mL/min; 214 om setector, temperate ambient, sensitiviy 0.L AUFS. ‘Cohuma sytem is ent factory when bseline separation is achieved ‘Deeweon sugar feat peak snd qunie acid peak in eranbemy juice cocktell andr apple juice Injct §-20 nL standard solution ater ‘ach 2 text lution injections to check linearity. Inject 5 lL est ‘BROW AOAC INTERNATION [ADAG OFFICAL METHODS OF ANALYSIS (2010) ° MIN 8 Figure 986.13, Liquid chromatogram of cranberry juice cocktail: (1) solvent and sugar tront; (2) quinic acid; (3) malic acid; (4) ascorbic acid and some shikimic acid; and (5) citric acid. solution. Use averge of injection fr standard and test soution responses. Eton ptr: quine, malic citi, fume, calevatons 7 (Quis, malls ce o fumare cid, %= (PUPA) XVID XC where PA and PA’ = peak area of test solution and standard, respectively, V and 1° = volun of test solution znd standard, espectively; and C= conceataton of standard, [Note Mali tandacde(Cemlic o O-L-malic will yield 2 pels, the second of whichis fanadie acid. Amountof fumarie ac present in mati aid standard i 0 small hat standard is 299.9% famatic feiéfice. Impurity does not affest analysis. Cranberry juice ‘cock, forfed with vita C, wil exhibit arge peak appearing SBermalic (Thisis due to ceelution of ecorbiescid+shikimic acid fad {3 not quantified) For dels about this separation see Figure 986.13) Reference: JAOAC 8, 5941986). CAS-77.92-9(ctie ace) CAS-6015-15-7 (wale acid) CAS-77.95-2 (quinie act) Resse: March 1996 aTAAT AOAC Official Method 937.10 Lactic Acid in Fruits and Fruit Products. ‘Spectrophetometie Method Fret Action 1937 inal Action ‘See 22.089, 1200 Ba. 371.48 AOAC Official Method 969.30 Organi Acids (Foreign) in Fruit Juices Paper Chromatographie Mathes First Action 1969 Final Action 1980 Caution: Sex Appendis Bele noes co hydogen sul.