| Abstract [eng] |
This study presents optimization of sirolimus and tacrolimus measurement in patients blood after renal transplantation by high–performance liquid chromatography with mass–spectrometry detection. In experiments with standart solutions of sirolimus and tacrolimus, using ascomycin as internal standart, chromatographical conditions were tested: type of eluation (isocratical and gradiental), properties of eluents (variable acidity of polar solvent and hydrophobity of nonpolar solvent), linearity of analysis. Patient blood extraction procedure was also tested. Sirolimus concentrations measured by HPLC-MS/MS method were 8-9 times lower as received by MEIA method. Both types of eluation were not sufficient for routine analysis in clinical laboratory, when eluents methanol and 0,2% acetic acid solvent were used. Gradiental eluation seamed to be better, but linearity analysis of sample series showed, that chromatographical system was not well-balanced. Methanol was changed to acetonitrile, but separation and retention time of analytes were even worse. Best results were obtained using mixture of methanol and tetrahydrofuran as nonpolar eluent. |
