AOAC SPIFAN Nutrients ERP Review (March 14, 2019)

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

(g) β-Carotene system suitability solution.— Transfer 20 mg β-carotene system suitability reference standard, D(f) , to a 50 mL volumetric flask. Add 1 mL water and 4 mL THF and sonicate 5 min. Dilute to volume with IPA and sonicate 5 min. Cool to room temperature, filter the cloudy suspension through 0.2 μm PTFE, and dilute the clear filtrate 1:4 with IPA. Store in glass vials in the refrigerator up to 3 months or at -20°C for up to 6 months. (h) Lutein system suitability solution.— Transfer 10 mg USP lutein, D(e) , to a 100 mL volumetric flask, dilute to volume with ROH, and mix. Transfer 1 mL to a 50 mL volumetric flask. Add ca. 35 mL IPA to the 50 mL flask, stopper loosely, and heat in a water bath at 80 °C for two hours. Cool to room temperature, add 250 μL apocarotenal stock solution E(a) with an adjustable pipette, and dilute to volume with IPA. Store in glass vials in the refrigerator up to 3 months or at -20°C for up to 6 months. (i) Internal standard solution (ISTD).— Prepare immediately before use. (1) For infant formula and samples with low carotenoid concentrations (up to 100 μg of an individual carotenoid per 100 g): Transfer 4.0 mL apocarotenal working solution, E(c) , to a 50 mL volumetric flask and dilute to volume with pyrogallol solution, C(o) . This is enough solution for 9 samples. (2) For samples with individual carotenoid concentrations >100 μg/100 g: Transfer 4.0 mL apocarotenal intermediate solution, E(d) , to a 50 mL volumetric flask and dilute to volume with pyrogallol solution. Note: The apocarotenal solutions used to make the ISTD should be made from the same apocarotenal stock solution, E(a) , as that used to make the calibration solutions, E(f) .

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