AOAC SPIFAN Nutrients ERP Review (March 14, 2019)

2016.13 (Jan. 2019) Carot-02 MLT Report FOR ERP USE ONLY DO NOT DISTRIBUTE

(c) Individual carotenoid working solutions for chromatographic purity check (200 μg/100 mL).— For lutein and β-carotene: With an adjustable pipette, transfer 100 μL of each standard stock solution, E(a) , to separate 10 mL volumetric flasks and dilute each to volume with sample solvent. For lycopene: With an adjustable pipette, transfer 200 μL standard stock solution, E(a) , to a 10 mL volumetric flask and dilute to volume with sample solvent. For apocarotenal: Transfer 1.0 mL standard stock solution, E(a) , to a 100 mL volumetric flask and dilute to volume with sample solvent. Store in glass vials at -20 °C for up to one month and use for internal standard, E(i) . Analyze by UHPLC on the same day prepared and calculate the chromatographic purity of each working solution according to section H(a) . (d) Apocarotenal intermediate solution (1200 μg/100 mL).— Transfer 3.0 mL apocarotenal stock solution, E(a) , to a 50 mL volumetric flask and dilute to volume with sample solvent. Store in glass vials at -20°C for up to 1 month. (e) Mixed carotenoid intermediate solution (400 μg/100 mL).— Combine 2.0 mL each of lutein, β-carotene, and lycopene standard stock solutions, E(a) , in a 100 mL volumetric flask and dilute to volume with sample solvent. Store in glass vials at -20°C for up to 1 month. (f) Calibration solutions.— Transfer apocarotenal intermediate solution, E(d) , and mixed carotenoid intermediate solution, E(e) , to volumetric flasks according to Table B and dilute to volume with sample solvent. Store in glass vials at -20°C for up to 1 month. Solutions maybe be aliquoted to HPLC vials prior to storing in the freezer.

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