Alternative LC-MS-MS Method for Simultaneous Determination of Proguanil, Its Active Metabolite in Human Plasma and Application to a Bioequivalence Study
2009
Pingale, Satish G | Nerurkar, Ketan K | Padgaonkar, Ajit M | Pawar, Uttam D | Mangaonkar, Kiran V
An alternative rapid and sensitive liquid chromatography-tandem mass spectrometry method has been developed and validated for simultaneous analysis of proguanil (PRO) and cycloguanil (CYC) in human plasma. The analytes were extracted from human plasma by solid phase extraction. Riluzole (RIL) was used as an internal standard for proguanil and cycloguanil. A HyPURITY Advance C18 column provided chromatographic separation of analytes followed by detection with mass spectrometry. The method involves simple isocratic chromatography conditions and mass spectrometric detection in the positive ionization mode using an API-4000 system. The proposed method has been validated with linear range of 1.5-150.0 ng mL⁻¹ for PRO and 0.5-50.0 ng mL⁻¹ for CYC. The inter-run and intra-run precision values are within 2.54, 9.19% for PRO and 1.99, 10.69% for CYC at LOQ levels. The overall recoveries for PRO and CYC were 102.52 and 106.72%, respectively. Total elution time was as low as 2.50 min. This validated method was used successfully for analysis of plasma samples from a bioequivalence study.
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