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LC-MS/MS Analysis of Nicotinic acid
and Nicotinamide
Chemistry Method
Version 2
Page 4/15
Prepared by
S. Bhandari
Approved by
Issue Date
Implementation Date
________________________________________________________________________________
Draft Date: 3/2/16 Copyright © Merieux NutriSciences, 2016
Uncontrolled When Printed
2.
Flow diagram
Weigh about 0.2g sample
10 mL Extraction Buffer and Sonicate 30 min
Add 0.1 mL protease solution & incubate at
60◦C for 30 min
Boil for 10 min and cool to RT
Add 1 mL Takadiastase solution and incubate
at 45◦C for 3 hrs
Boil for 5 min and cool to RT and mix water
Bring to Final Volume of 50 mL with extraction
buffer
Take an aliquot of 1 mL of the extract in LC vial ,
& add 30 uL of mixed internal std. solution
Analyze by LC-MS/MS
Filter an aliquot through 0.45 micron PVDF
folter
METHOD
FOR ERP USE ONLY
DO NOT DISTRIBUTE