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Abbott Nutrition Division
Simultaneous Determination of Total Vitamin B
6,
B
2,
B
3
and B
1
in Infant
Formula Products by LC-MS/MS Using Enzymatic Digestion
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
Column
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 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
Methanol
Strong Wash – Volume
600 µL
Weak Wash – Solvent
Water
Weak Wash - Volume
1000 µL
Gradient Program
Time
% A1
% B1
Curve
0.0
99.0
1.0
6
1.5
99.0
1.0
6
1.6
75.0
25.0
6
3.5
75.0
25.0
6
3.6
1.0
99.0
6
5.0
1.0
99.0
6
5.1
99.0
1.0
6
7.0
99.0
1.0
6
27
BVit-02
FOR ERP USE ONLY
DO NOT DISTRIBUTE