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Clin Biochem. 2008 Jun;41(9):728-35. doi: 10.1016/j.clinbiochem.2008.02.014. Epub 2008 Mar 12.

Simultaneous quantification of cyclosporine, tacrolimus, sirolimus and everolimus in whole blood by liquid chromatography-electrospray mass spectrometry.

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1
Laboratory Medicine Service, University Hospitals of Geneva, Geneva, Switzerland.

Abstract

OBJECTIVES:

The aim of this work was to develop a selective method for the simultaneous quantification of cyclosporine, tacrolimus, sirolimus and everolimus in whole blood.

DESIGN AND METHODS:

An automated on-line solid-phase extraction system coupled with liquid chromatography-mass spectrometry (LC-MS) was used. After a simple protein precipitation, the supernatant was load on a C8 column with a mobile phase composed of MeOH/H(2)O (5/95 v/v), supplemented with formic acid 0.02% and sodium formate 1 muM. After column-switching, the analytes were transferred in the back-flush mode on a C18 column with MeOH/H(2)O (65/35). The valve was then commuted to its initial position and the chromatographic separation was performed with a gradient of MeOH/H(2)O (65/35-95/5). The sodium adducts [M+Na](+) were monitored for quantification with an electrospray ionization-single quadrupole MS.

RESULTS:

The LC-MS assay was fully validated on a concentration range of 2.5-30 ng/mL for tacrolimus, sirolimus and everolimus and of 50-1500 ng/mL for cyclosporine, allowing a quantification of cyclosporine 2 h post-dose without sample dilution. Trueness, repeatability and intermediate precision were found to be satisfactory.

CONCLUSION:

This method provided a selective, rapid and automated procedure that can be easily used for routine quantification of immunosuppressive drugs in most clinical laboratories.

[Indexed for MEDLINE]

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