Development and validation of a liquid chromatography/tandem mass spectrometry method for determination of caspofungin in dried blood spots.

Rationale: A liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for quantification of caspofungin in dried blood spots (DBS) was developed and validated. Methods: The DBS samples were prepared by spotting whole blood onto Whatman 903 filter paper, drying at room temperature and extracting with 50% methanol and further cleaned by protein precipitation with acetonitrile. Roxithromycin was selected as internal standard, and the separation of the analytes with endogenous ingredients was accomplished on a Hypersil GOLD aQ column with a mobile phase composed of 0.1% formic acid (v/v) and methanol in gradient mode. The detection of the analytes was performed on a triple quadrupole mass spectrometer in positive electrospray ionization mode, and the following selective reaction monitoring (SRM) transitions were monitored: m/z 547.6 → 538.7 and 837.4→ 679.4 for quantification of caspofungin and the internal standard, respectively. Results: The total analytical time was 8 min for each run. The calibration curve exhibited a good linearity over the range from 0.2 to 20 μg/mL and the lower limit of quantification (LLOQ) was 0.2 μg/mL for caspofungin in DBS. The recoveries of caspofungin ranged from 62.64% to 76.69%, and no obvious matrix effect was observed. The intra‐ and inter‐day precision and accuracy were within acceptable limits, and caspofungin in DBS was stable after storage at room temperature for 24 h and at −80°C for 30 days. There was no evident effect of the hematocrit value on the analysis of caspofungin. Conclusions: The proposed method presents an alternative to the conventional venous sampling method, and was successfully utilized for pharmacokinetics study of caspofungin in ICU patients. [ABSTRACT FROM AUTHOR]