AOAC SPIFAN Nutrients ERP Review (March 14, 2019)

2016.13 (SEPTEMBER 2017) CAROT-02 VALIDATION REPORT FOR ERP USE ONLY DO NOT DISTRIBUTE

Table 5. LOD and LOQ for lutein in spiked SRM 1849a.

Measured conc (mcg/100 g as fed)

Extrapolated LOD (mcg/100 g)

Extrapolated LOQ (mcg/100 g)

Sample

S/N 68.6 70.4 37.0 Avg

1 2 3

1.40 1.46 1.43

0.06 0.06 0.12 0.08

0.20 0.21 0.39 0.27

Table 6. LOD and LOQ for β-carotene in spiked SRM 1849a.

Extrapolated LOD (mcg/100 g)

Extrapolated LOQ (mcg/100 g)

Measured conc (mcg/100 g as fed)

Sample

S/N

1 2 3

1.62 1.70 1.66

61.6 61.2 61.0

0.08 0.08 0.08 0.08

0.26 0.28 0.27 0.27

Avg

Table 7. LOD and LOQ for lycopene in spiked SRM 1849a.

Measured conc (mcg/100 g as fed)

Extrapolated LOD (mcg/100 g)

Extrapolated LOQ (mcg/100 g)

Sample

S/N 48.2 48.3 47.4 Avg

1 2 3

1.43 1.46 1.40

0.09 0.09 0.09 0.09

0.30 0.30 0.29 0.30

Specificity

The SMPRs call for the determination of all- trans and cis isomers of lutein and β-carotene, as well as separation of lutein from zeaxanthin. Specificity was evaluated with visual inspection of chromatograms and by measuring resolution of system suitability standard mixtures. Because apocarotenal is used as an internal standard, samples were prepared without internal standard to ensure there were no interfering peaks. Standard mixtures were isomerized with heat to identify major cis isomers. Peak assignments were based on relative retention times from previous studies using C30 columns and methanol/MTBE as the mobile phase (Aman et al., 2005; Marx et al, 2000; Ping and Lian, 2006; Puspitasari-Nienaber, 2002). Samples spiked with all- trans -lutein and