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Clinical Chemistry 53: 1827-1834, 2007. First published August 16, 2007; 10.1373/clinchem.2007.090811
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(Clinical Chemistry. 2007;53:1827-1834.)
© 2007 American Association for Clinical Chemistry, Inc.

Automation and Analytical Techniques

Methanol-Associated Matrix Effects in Electrospray Ionization Tandem Mass Spectrometry

Thomas M. Annesleya

Department of Pathology, University of Michigan Health Sciences Center, Ann Arbor, MI.

aAddress correspondence to the author at: University Hospital, Rm. 2G332, 1500 East Medical Center Dr., Ann Arbor, MI 48109-5054. Fax 734-763-4095; e-mail annesley{at}umich.edu.

Background: Matrix effects can profoundly reduce the performance of electrospray ionization mass spectrometry. Preliminary observations indicated that the methanol used in the mobile phase could be a source of differential ionization or ion suppression.

Methods: Drug stability studies, analysis of biological extracts, mixing experiments, and postcolumn infusions were used to test 9 commercial methanols for ionization differences in liquid chromatography-tandem mass spectrometry assays for immunosuppressants. Area responses for the drugs and internal standards were compared for mobile phases prepared with each selected methanol. Postcolumn infusion experiments were performed to confirm the degree of ionization differences occurring at the ion source, and to evaluate the proportions of ammonium, sodium, and potassium adducts.

Results: The decrease in signal for the immunosuppressant drugs was shown to result from differential ionization associated with the selected methanols. Product ion intensity varied by 10-fold among the methanols tested. For sirolimus, tacrolimus, and mycophenolic acid, the percentage change in ionization was the same for the drug and its corresponding internal standard. Postcolumn sirolimus infusion evaluation revealed that a 1000-fold analyte concentration difference did not affect ionization. The proportions of ammonium, sodium, and potassium adducts of sirolimus precursor ions differed in relation to the source of methanol.

Conclusions: Organic solvents used in mobile phases and extract preparation of biological samples may be associated with ion suppression, affecting adduct formation and assay sensitivity.




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