Review Team (February 2016)

Amino-03 (February 2016) FOR ERP USE ONLY DO NOT DISTRIBUTE

Amino Acids in Infant Formula and Adult/Pediatric Formulas Ultra High Performance Liquid Chromatography

Column

ACCQ-TAG ULTRA C18 1.7μM, 2.1 x 100mm

Mobile phase A Mobile phase B

Mobile Phase A1 Mobile Phase B1 0.6 mL/minute

Flow rate

Injection Volume

1.0 μL 50 °C 260 nm

Column Temperature Detector Wavelength

Run Time

14 minutes Gradient Method

Time (minutes)

A %

B%

0.00 0.63 2.33 2.92 3.50 4.12 4.70 5.64 5.99 6.16 6.39 6.70 9.03

99.90 99.90 99.50 98.70 98.00 97.00 96.00 93.90 93.10 92.60 92.00 90.90 78.80 40.40 10.00 10.00 99.90 99.90

0.10 0.10 0.50 1.30 2.00 3.00 4.00 6.10 6.90 7.40 8.00 9.10

21.20 59.60 90.00 90.00

10.00 10.01 11.01 11.02 14.00

0.10

0.10 3) Chromatographic conditions for Alkaline Hydrolysis method: Table 8: Column

Agilent Zorbax RRHD, 1.8μm, 2.1 X 50 mm

Mobile phase

Mobile Phase A2 0.2 mL/minute

Flow rate

Injection Volume

2.0 μL

Column Temperature Detector Wavelength

25°C

Ex. 280nm, Em. 356nm

Run Time

10minutes

Pump Mode

Isocratic

7