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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
Thiamin
250
Riboflavin
800
QS to the mark with laboratory water.
Expiration: Prepare fresh daily.
i)
Working Standard Solutions
NOTE: As a general rule, use the Gilson pipette with the smallest maximum volume to
dispense the specified volumes of IWSS and ISSM solutions.
Combine the IWSS and ISSM solution volumes shown in Table 8 into a 50-mL self-standing
centrifuge tube. Pipette in 5 mL of enzyme solution, and then add 50mM Ammonium Formate up
to a final volume of 30mL.
Expiration: Prepare fresh daily.
Table 5. Working Standard solution preparation.
Solution
IWSS
Volume
(µL)
Gilson Pipette
(recommended)
ISSM
Volume
(µL)
Gilson Pipette
(recommended)
Final
Volume
(mL)
WS 1
25
M25
50
M100
30
WS 2
50
M100
50
M100
30
WS 3
100
M100
50
M100
30
WS 4
250
M250
50
M100
30
WS 5
500
M1000
50
M100
30
WS 6
1000
M1000
50
M100
30
.
NOTE: The accurate and precise addition of the IWSS and the ISSM solutions are critical
to the accuracy and precision of the method. Extreme care must be taken in
aliquoting the IWSS into the standards and the ISSM into the standards and the
samples.
NOTE: Due to the addition of internal standards for each compound the final volume of
the working standards is not a critical volume. Small errors (on the order of
approximately 1-2 mL) in the final volume will not adversely affect the calibration
accuracy.
36
BVit-02
FOR ERP USE ONLY
DO NOT DISTRIBUTE