AOAC SPIFAN Review Team Meeting Book-July 22, 2015

BVIT-02 FOR ERP/WORKING GROUP USE ONLY DO NOT DISTRIBUTE

Abbott Nutrition Division Simultaneous Determination of Total Vitamin B 6, B 2, B 3 in Infant Formula Products by LC-MS/MS Using Enzymatic Digestion and B 1

9. Filter an aliquot of the sample extract into an amber autosampler vial using a 0.20 µm PTFE syringe filter. NOTE: Ensure that the filtered sample solution is clear. If the filter fails the solution in the autosampler vial will appear cloudy. If this is observed discard this sample and filter another aliquot of the sample into a clean autosampler vial. Samples are stable for up to 48 hours at room temperature. Working standard solutions and samples must be tested as a complete set if reanalysis is required. It is recommended that the autosampler vials be recapped if reanalysis is anticipated. CALCULATIONS AND EXAMPLES 1. All calculations are performed using MassLynx software, current revision (Version 4.1, or later) fitted with Target Lynx data processing software. 2. Analyte concentrations in product samples are calculated by comparison of calculated response ratios for the analyte in the sample to that calculated for the calibration standards using least squares regression. Response ratios used for the linear regression are calculated using the ratio of the response of the analyte to that of its internal standard. This ratio is then plotted against the concentration of the standard.

F.

3. Vitamin Stock Solutions – Native Stock Solutions [ ] Vol Vit s =

PSMW 000 ,1 ××× ×

Where:

[Vit]: Vitamin Standard Solution Concentration, in m g/mL W S : Weight of Standard, in mg S: Stoichiometric correction factor (0.8227, 0.8209, and 0.6976 for Pryridoxine, Pyridoxal, and Pyridoxamine respectively). M: Moisture content correction factor for the Standard (per AN-RS Method 1.57), (used for Thiamin)

) (% 100 Moisture − −

M

=

100

P: Purity of standard as defined by the manufacturer 1,000: Units Conversion Factor, mg to µg Vol: Solution Volume, in mL

a) Pyridoxine (PN) Stock Solution Concentration Calculation

W

P

8227 .0

000 ,1

×

××

[ ] PN

PM W

PN

10 ×× × =

=

S

PN

mL

100

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