AOAC SPIFAN Nutrients ERP Review (March 14, 2019)

Carot-02 (February 2016) FOR ERP USE ONLY DO NOT DISTRIBUTE

Table 1. Calibration error for each carotenoid. all- trans -Lutein

all- trans -β-Carotene

Apocarotenal

Conc (μg/100 mL)

Conc (μg/100 mL)

Conc (μg/100 mL)

Residual

Residual

Residual

185.6

0.427848006 1.169099574 -4.622149486 1.855899172 -4.881101079

244.0 122.0

-1.561049661 4.765184349 -3.505075245 -1.581828442 1.882768999

257.0 128.5

1.663495109 -1.823403562 -3.373542764 -1.846922498 5.380373715

92.8 37.1 18.6 3.71

48.8 24.4 4.88

51.4 25.7 5.14

C. Accuracy – Accuracy was determined by spike and recovery from SRM 1849a. SRM 1849a is not fortified with lutein or β-carotene, but contains very low levels of β-carotene.

Table 2. Recovery of lutein and β-carotene spiked into SRM 1849a (n=3). Background levels of β- carotene were taken into account. all- trans -Lutein all- trans -β-Carotene Spike level (μg/100 g as fed) 12.9 1.29 15.3 1.53 Avg recovery 99.0% 100.2% 101.9% 96.5% %RSD 0.05% 0.18% 0.51% 0.62%

D. Precision Table 3. Repeatability of milk-based infant formula from SPIFAN sample set (n=3).

cis -Lutein all- trans -Lutein cis -β-Carotene all- trans -β-Carotene

Avg (μg/100 g as fed)

ND n/a

4.5

6.7

13.6

%RSD

0.27%

0.68%

0.29%

Table 4. Repeatability of PN soy-based infant formula (n=3).

cis -Lutein all- trans -Lutein cis -β-Carotene all- trans -β-Carotene

Avg (μg/100 g as fed)

ND n/a

4.1

7.3

15.2

%RSD

0.58%

1.04%

0.21%

E. LOQ – Based on spike and recovery data a LOQ of 1.3 μg/100 g was observed for lutein and 1.5 μg/100 g for β-carotene. F. References (a) Aman et al. (2005). Food Chem . 92: 753-763.

(b) Craft and Soares (1992). J. Agric. Food Chem . 40: 431-434. (c) Granado et al. (2001). J. Food Comp. Anal . 14: 479-489. (d) Marx et al. (2000). Food Chem . 70: 403-408. (e) Müller et al. (2008). J. AOAC Int . 91(6): 1284-1297.

(f) Puspitasari-Nienaber et al. (2002). JAOCS 79(7): 633-640. (g) Szpylka and DeVries (2005). J. AOAC Int . 88(5): 1279-1291. (h) The United States Pharmacopeial Convention (2014). Revision Bulletin - Beta Carotene. (i) Waysek et al. (2010). J. AOAC Int . 93(2): 499-509.