SPIFAN Nutrients ERP Book_9-29-15

Amino-02 FOR ERP USE ONLY DO NOT DISTRIBUTE

HPLC Determination of Total Tryptophan in Infant Formula and Adult/Pediatric Nutritional Formula Following Enzymatic Hydrolysis

Abbott Nutrition, Division of Abbott Laboratories

1. Accuracy: a) Average overspike recoveries of Hypoallergenic Infant formula with BSA and free tryptophan (n=10): 1) NIST BSA: 99.1% 2) Free Tryptophan: 101.3% a) NIST 1849a assay percent of reference value: 100.4% (mean of n=10)

2. Precision:

a) NIST 1849a (n=30 over 6 months): 2.14% b) RSD for ten independent runs of duplicate analyses.

SAMPLE

Within run

Run-to-run

Total RSD

Soy Formula

0.48% 0.62%

1.64% 1.68%

1.71% 1.79%

Hypoallergenic Formula

3. Second Laboratory Performance with NIST 1849a (n=14) a) RSD 0.99%

b) Mean value: 0.186 g/100g (101.1% of Reference Value)

J. FIGURES

FIGURE 1 Typical Middle Standard Chromatogram

15.00

14.00

13.00

12.00

TRP_SA - 2.395

11.00

5MDLT - 6.035

10.00

9.00

8.00

LU

7.00

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3.00

2.00

1.00

0.00

0.50

1.00

1.50

2.00

2.50

3.00

3.50

4.00

4.50

5.00

5.50

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6.50

7.00

Minutes

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