AOAC SPIFAN Nutrients ERP Review (March 14, 2019)

2016.13 (SEPTEMBER 2017) CAROT-02 METHOD

FOR ERP USE ONLY DO NOT DISTRIBUTE (o) Pyrogallol solution (0.2 M pyrogallic acid in ethanol).— Dissolve 6.3 g pyrogallic acid in 250 mL ROH. Store in refrigerator up to 1 month. Solution should be clear at room temperature; discard if colored. (p) Extraction solution (10 mM vitamin E in 1:1 MTBE/THF).— Dissolve 2.2 g α-tocopherol in 250 mL MTBE and 250 mL THF. Store in refrigerator up to 1 month. (q) Sample solvent (10 mM vitamin E in IPA).— Dissolve 4.4 g α-tocopherol in 1000 mL IPA. Store in refrigerator up to 1 month. (r) Mobile phase for LC system.—(1) Phase A .—20 mM ammonium acetate in methanol– water (98+2). Combine 980 mL MeOH, 20 mL water, and 1.54 g ammonium acetate and mix to dissolve. (2) Phase B .—MTBE. D. Standards (a) Lutein .—ChromaDex (Irvine, CA) Part No. ASB-00012453 or equivalent. (b) β-Carotene .—USP (Rockville, MD) Part No. 1065480 or equivalent. (c) Apocarotenal (β-Apo-8′-carotenal) .—USP Part No. 1040854 or equivalent. (d) Lycopene . – ChromaDex Part No. ASB-00012550 or equivalent (e) Lutein containing ca. 10% zeaxanthin .—USP Part No. 1370804. (f) β-Carotene system suitability reference standard .—USP Part No. 1065491. E. Standards Preparation Standard solution preparation is summarized in Table A and detailed below. (a) Carotenoid stock solutions (20 000 μg/100 mL).— Weigh (to 0.01 mg) approximately 10 mg each of lutein, D(a) ; β-carotene, D(b) ; and apocarotenal, D(c) reference standard into separate 50 mL volumetric flasks. Add ca. 40 mL MTBE/Vitamin E, C(m) , to each, sonicate 2-3 min, and dilute to volume with MTBE/Vitamin E. Weigh (to 0.01 mg) approximately 5 mg each of lycopene, D(d) reference standard into a 50 mL volumetric