AOAC SPIFAN Review Team Meeting Book-July 22, 2015

BVIT-02 FOR ERP/WORKING GROUP USE ONLY DO NOT DISTRIBUTE

Abbott Nutrition Division Simultaneous Determination of Total Vitamin B 6, B 2, B 3 in Infant Formula Products by LC-MS/MS Using Enzymatic Digestion and B 1

f) Data Processing: Waters MassLynx 4.1 (or later version) with TargetLynx data processing package. g) Nitrogen Generator: Peak Scientific Nitrogen Generator; Model # NM30LA, or equivalent.

3. Instrument Conditions a) UPLC Instrument Configuration and Operating Parameters

Table 1. UPLC instrument configuration details (Injection 1). Mobile Phase A1 20 mM Ammonium Formate Mobile Phase B1 Methanol Flow Rate 250 µL/min Flow Rate into MS 250 µL/min Degasser On

Waters Acquity UPLC BEH C-18 1.7 µm 2.1 x 50 mm with a Waters BEH C18 1.7 µm 2.1 x 5mm Vanguard Column

Column

Column Temperature

40 ° C

Injection Type

10 µL full loop injection with 10 µL sample loop installed

Loop Volume Overfill

Automatic

Sample Temperature

25 ºC

Injector Wash Settings Strong Wash - Solvent Strong Wash – Volume Weak Wash – Solvent Weak Wash - Volume Gradient Program

Methanol

600 µL

Water

1000 µL

Time

% A1

% B1

Curve

0.0 1.5 1.6 3.5 3.6 5.0 5.1 7.0

99.0 99.0 75.0 75.0

1.0 1.0

6 6 6 6 6 6 6 6

25.0 25.0 99.0 99.0

1.0 1.0

99.0 99.0

1.0 1.0

27