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Simultaneous Determination of Thiamine HCl, Riboflavin, Niacin (Nicotinic Acid and Nicotinamide),
Pantothenic acid, Vitamin B6 (Pyridoxine, Pyridoxal and Pyridoxamine) and Biotin in Infant
Formula and Adult Nutritionals
CONFIDENTIAL METHOD FOR ERP ASSESSMENT ONLY
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Typical LC Time
Program
Step
Total Time
(min)
Flow rate
(µL/min)
A (%)
B(%)
0
0.00
400
100
0
1
0.05
400
100
0
2
1.50
400
85
15
3
5.00
400
80
20
4
6.50
400
10
90
5
8.50
400
10
90
6
9.00
400
100
0
7
12.00
400
100
0
Stop Time
12 minutes
Retention Time
Nicotinic acid approximately 2.0-2.5 minutes
Nicotinamide approximately 2.0-2.5 minutes
Pyridoxal approximately 2.8-3.3 minutes
Pantothenic acid approximately 2.8-3.3 minutes
Pyridoxamine approximately 2.8-3.3 minutes
Pyridoxine approximately 3.1-3.6 minutes
Thiamin hydrochloride approximately 4.0-4.5 minutes
Biotin approximately 6.0-6.5 minutes
Riboflavin approximately 6.3-6.8 minutes
D. System Suitability Requirements
a. Plot calibration between Concentration (ng/ml) and Area ratio
≥ 6 points
. The correlation coefficient of
each calibration
should be not less than 0.995. % Error of concentration should be ≤ 10%
.
b. When stable baseline is obtained, inject a middle level of working standard three times in succession
and determine the %RSD, The %RSD should be not more than 3%.
c. Analyte a middle level of working standard between 10 to 12 samples. % Error od centration should be
≤ 10%
d. For the confirmation, ion ratio should be with in 20%
B-VIT-01
FOR ERP/WORKING GROUP USE ONLY
DO NOT DISTRIBUTE
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