AOAC SPIFAN Nutrients ERP Review (March 14, 2019)

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

Summary The SPIFAN Expert Review Panel selected AOAC First Action Method 2016.13 to move forward to multi-laboratory testing (MLT) in August 2017 based on results of the single laboratory validation (SLV). This report details the MLT data. An LC-UV method for determination of lutein, β-carotene, and lycopene in infant formula and adult nutritionals was validated using the SPIFAN sample kit, including both unfortified and fortified samples. The sample set included blind duplicates of 6 matrices fortified with lutein, 7 matrices fortified with β-carotene, and one fortified with lycopene. The NIST SRM 1869 was included in the sample set as a reference material. Repeatability for all- trans -lutein ranged from 1.5 to 10.0% RSD and repeatability for total lutein ranged from 2.2 to 12.0% RSD. The repeatability values met the criteria in the SMPR for lutein with the exception of milk-based infant formula. Reproducibility for all- trans -lutein ranged from 10.5 to 14.8% RSD and for total lutein it ranged from 11.5 to 19.9% RSD. Although the all- trans -lutein data met the reproducibility limit for the SMPR, the data for total lutein data was above the SMPR limit for most matrices. All HorRat values for all- trans -lutein and total lutein are below 1.0, indicating that the method reproducibility is better than predicted based on historical data. One of the SPIFAN matrices fortified with β-carotene (elemental infant formula) was later determined to be non-homogeneous. Excluding this matrix, the repeatability for all- trans -β- carotene ranged from 2.0 to 4.2% RSD and repeatability for total β-carotene ranged from 1.1 to 6.0% RSD. Reproducibility for all- trans -β-carotene ranged from 7.4 to 15.3% RSD and for total β-carotene it ranged from 8.5 to 13.7% RSD. These data for all- trans - and total β-carotene met the repeatability and reproducibility limits in the SMPR. Repeatability for total lycopene was 1.6% RSD and reproducibility was 7.4% RSD. The repeatability and reproducibility values met the criteria in the SMPR for total lycopene.

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