AOAC SPIFAN Nutrients ERP Review (March 14, 2019)

2016.13 (SEPTEMBER 2017) CAROT-02 METHOD

FOR ERP USE ONLY DO NOT DISTRIBUTE The calibration and calculation may be achieved through data processing within the instrument software or off-line. (e) Calculate the mass of apocarotenal (M A ), in μg, added to the test samples: M A = (C A × V A ) × (4/50) where C A = the concentration of apocarotenal in the intermediate or WS used in the ISTD solution (μg/100 mL); V A = the volume of ISTD solution added to each sample, in mL; 4 = the volume of apocarotenal intermediate or WS used in the ISTD solution, in mL; 50 = the total volume of ISTD solution made, in mL. (f) Calculate the contents of all- trans lutein, cis isomers of lutein, and total lutein in the test samples. For peak identification, refer to relative retention times of peaks in Figures A , C , and D . Lut trans = (M A /M S ) × ([A Lut /A A ] – I Lut ) × (100/RF Lut ) where Lut trans = the all- trans lutein in the sample (μg/100 g); M A = the mass of apocarotenal added to the test sample, in μg; M S = the sample weight, in g; A Lut = the peak area of all- trans -lutein in the sample chromatogram (AU); A A = the peak area of apocarotenal in the sample chromatogram (AU); I Lut = the y-intercept of the calibration curve for all- trans -lutein; RF Lut = the slope of the calibration curve for all- trans -lutein. Lut cis = (M A /M S ) × ([(A 13cisLut + A 13’cisLut + A 9cisLut + A 9’cisLut )/A A ] – I Lut ) × (100/RF Lut ) where Lut cis = the cis isomers of lutein in the sample (μg/100 g); M A = the mass of apocarotenal added to the test sample, in μg; M S = the sample weight, in g; A 13cisLut = the peak area of 13- cis lutein in the sample chromatogram (AU); A 13’cisLut = the peak area of 13′- cis lutein in the sample chromatogram (AU); A 9cisLut = the peak area of 9- cis lutein in the sample chromatogram (AU); A 9’cisLut = the peak area of 9′- cis lutein in the sample chromatogram (AU); A A = the peak area of apocarotenal in the sample chromatogram