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

MassLynx main page. Shutdown the Acquity control panel, the inlet editor, the tune page and finally the MassLynx main window. Run the program “Stop Acquity Process” from the PC desktop. Once the program completes launch MassLynx and allow the software to fully load. If upon startup MassLynx does not require a security login or comes up with a sample queue running the PC will need to be rebooted to clear the error. From the main window of MassLynx, launch the Tune and Inlet editors. From the desktop, launch the Acquity Console. Once all four process windows are loaded verify that the tune page and Acquity console show the system ready for analysis. The sample list can then be restarted. 2. Chromatography a) Column Stability i) Previous experience has indicated that the BEH C-18 column is stable for approximately 2000 injections. ii) The column should be replaced when peak shape (e.g. tailing, fronting, of broadening) or retention times indicate deteriorating performance. b) Pyridoxine i) Pyridoxine normally elutes at approximately 1.47 minutes. The retention of Pyridoxine is readily affected by changes in mobile phase composition. If the acid concentration in mobile phase A is incorrect the retention time of Pyridoxine will shift significantly. ii) Pyridoxine can also be used as a good indicator of column performance. Past experience has demonstrated that as the column begins to fail the retention of Pyridoxine shifts earlier from ~ 1.5 minutes to ~1.2 minutes. 3. Mass Spectrometry a) Sensitivity Data review should include monitoring of the signal to noise (S/N) of the low working standard of each compound. Pyridoxamine and Pyridoxal have the weakest signal of the compounds being analyzed. The S/N for both compounds should be greater than 20. If routine cleaning of the source fails to restore system performance contact the system administrator or Waters service. b) Signal Saturation Waters triple quadrupole mass spectrometers have a maximum signal intensity of 1.34e 8 in SRM analysis. A signal that approaches this level may be saturating the detector and not producing a true response for the injected concentration. Evaluation of the high standards and samples should be conducted for samples with intensities greater than 1.0e 8 . These injections should be evaluated for peak shape (flat topped) and residuals of standards to ensure that the detector is not at saturation. 4. Quantitation a) Response Ratio Evaluation i) Care should be taken to review the response ratio for all compounds for the sample to ensure that the analyte concentration for the analytical sample is within the concentration range of the

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