AOAC SPIFAN Nutrients ERP Review (March 14, 2019)

2016.13 (SEPTEMBER 2017) CAROT-02 METHOD

FOR ERP USE ONLY DO NOT DISTRIBUTE

(e) Shake on reciprocating shaker 5 min. (f) Add 8 mL extraction solution, C(o) , to each tube with a repeater pipet. (g) Shake 10 min. (h) With a repeater pipet or dispenser, add 10 mL water and 10 mL hexane to each tube. (i) Shake 1 min. (j) Centrifuge at 1000 rpm (or equivalent to 200 × g ) for 5 min. (k) Transfer a portion of the supernatant to a 12 mL scintillation vial. (1) Samples with individual carotenoid concentrations ≤50 μg/100 g – use 10 mL supernatant (2) Samples with individual carotenoid concentrations >50 μg/100 g – use 3 mL supernatant (l) Dry under nitrogen at ≤40 °C. (m) Reconstitute dried extract in sample solvent and vortex to mix. (1) Samples with individual carotenoid concentrations ≤100 μg/100 g – add 0.5 mL. (2) Samples with individual carotenoid concentrations 100-500 μg/100 g – add 1 mL. (3) Samples with individual carotenoid concentrations 500-1000 μg/100 g – add 2 mL. (4) Samples with individual carotenoid concentrations 1000-1500 μg/100 g – add 3 mL. (n) Filter through 0.2 μm PTFE syringe filter prior to injection. G. Chromatography (a) Chromatographic conditions.— Set up the UHPLC system according to the specifications in Table C . Follow the manufacturer’s instructions for column installation, cleaning, and storage. (b) System suitability checks.—