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LC-MS/MS Analysis of Nicotinic acid
and Nicotinamide
Chemistry Method
Version 2
Page 10/15
Prepared by
S. Bhandari
Approved by
Issue Date
Implementation Date
________________________________________________________________________________
Draft Date: 3/2/16 Copyright © Merieux NutriSciences, 2016
Uncontrolled When Printed
1.
Use HPLC as described in the Apparatus section.
2.
Detetor Mass spectrometer, API5500 Qtrap or 6500 Qtrap
3.
Column:
Thermo Acclaim C30 3um 2.1 x 150mm
4.
HPLC Column Temperature: 30°C
5.
Autosampler Temperature: 4-5°C
6.
Injection Volume: 2 µL
7.
Mobile Phase: A:
8.
M.P.A: 12 mM ammonium formate (Optima) in water pH = 3.0
9.
M.P.B: 100% ACN
10.
Instrument HPLC Conditions
11. Dionex Ultimate 3000 / Agilent 1290 System
12.
(
a
)
Columns.—
Thermo Acclaim C30 3um 2.1 x 150mm
13.
(
b
)
Mobile Phase. —
14. M.P.A: 12 mM ammonium formate (Optima) in water pH = 3.0
15. M.P.B: 100% ACN
16.
(
c
)
Gradient. —
#
Time (min)
Flow Rate
(uL/min)
A%
B%
0
0.0
300
100
0
1
4.0
600
100
0
2
11.5
600
64
36
3
12.0
600
0
100
4
15.5
600
0
100
5
16.0
600
100
0
6
21.0
600
100
0
METHOD
FOR ERP USE ONLY
DO NOT DISTRIBUTE