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LC-MS/MS Analysis of Nicotinic acid
and Nicotinamide
Chemistry Method
Version 2
Page 12/15
Prepared by
S. Bhandari
Approved by
Issue Date
Implementation Date
________________________________________________________________________________
Draft Date: 3/2/16 Copyright © Merieux NutriSciences, 2016
Uncontrolled When Printed
B3-amide_Qual1
123.3
78.1
20
45
10
34
10
B3-amide_Qual2
123.3
53
20
45
10
35
10
B3-acid IS1
128.1
83
20
50
10
28
10
B3-acid IS2
128.1
54
20
50
10
29
10
B3-acid IS3
128.1
81
20
50
10
28
10
B3-amide IS1
127
80
20
45
10
27
10
B3-amide IS 2
127
82
20
45
10
34
10
B3-amide IS 3
127
55
20
45
10
35
10
31. MS
_Start.OnDuration=0.5
32. MS_Start.Off Duration=12.00
4.9 Calculations and reporting:
4.9.1 Preparation of Calibration Curve:
1. Draw calibration curve of each analyte using the following relation.
X axis = ratio of ng/mL of the analyte vs concentration (ng/mL) of the internal std.
Y axis = ratio of peak area of the analyte vs the peak area of the respective internal std.
. Linear regression is
performed. By the MS software.
4.9.2. Calculation of each of the analytes in the samples, Blank and Std. as a
sample:
1.
Determine of peak area of the analyte vs the peak area of the respective internal std. for
each sample extract and std. as a sample solution.
2.
Determine ng/mL of the analyte in each sample extract std. as a sample solution.
3.
Calculate ng/mL of each analyte in sample extracts as well as blank and
std. as a sample solutions. based on the respective calibration curve.
4.
Adjust analyte concentration in sample extract and std. as a sample
solution by deducting respective analyte concentration estimated in the
blank.
METHOD
FOR ERP USE ONLY
DO NOT DISTRIBUTE