Determination of Lutein, β-Carotene, and Lycopene in Infant Formula and Adult Nutritionals by Ultra-High Performance Liquid Chromatography: Collaborative Study, Final Action 2016.13 for β-Carotene and Lycopene Only

Background: Lutein, β-carotene, and lycopene are among the most common carotenoids present in human milk and are frequently added to infant formula and adult nutritionals.

Objective: A collaborative study was conducted to assess the interlaboratory performance of AOAC Official MethodSM2016.13 for the determination of lutein, β-carotene, and lycopene in infant formula and adult nutritionals.

Methods: Thirteen laboratories agreed to participate in the study and 10 laboratories from seven different countries reported results. The study samples included blind duplicates of 6 matrices fortified with lutein, 7 matrices fortified with β-carotene, and 1 fortified with lycopene. NIST SRM 1869 was included in the sample set as a reference material.

Results: After the removal of outliers and invalid data, the repeatability (RSDr) data was ≤10.0% for all-trans-lutein, ≤12.0% for total lutein, ≤4.2% for all-trans-β-carotene, ≤6.0% for total β-carotene, and 1.6% for total lycopene. Reproducibility (RSDR) was ≤14.8% for all-trans-lutein, ≤19.9% for total lutein, ≤15.3% for all-trans-β-carotene, ≤13.7% for total β-carotene, and 7.4% for total lycopene.

Conclusions: The repeatability and reproducibility values met the criteria in the Standard Method Performance Requirements (SMPRs) for β-carotene and lycopene and it was recommended that the method be approved as a Final Action for these analytes. Since the method did not meet the SMPR for lutein, it was recommended that it remain a First Action method for this analyte.

Highlights: AOAC Official MethodSM2016.13 was validated through a collaborative study to be accurate and reproducible for the determination of β-carotene and lycopene in infant formula and adult nutritionals.