Xây dựng phương pháp định lượng leuprolid trong huyết thanh chó bằng Phương pháp LC-MS/MS

Tóm tắt

Studying leuprolide pharmacokinetics using the LC-MS/MS technique to overcome selective and sensitive detection. This study aimed to develop and validate a high throughput and specific method using liquid chromatography tandem mass spectrometry to determine leuprolide in dog serum. Leuprolide and internal standard (paracetamol) were prepared from dog serum using protein precipitation followed by liquid-liquid extraction. The prepared samples were chromatographed on a column (50 x 2.1mm, i.d., 1.7 µm) with isocratic elution at a °ow rate of 0.2 ml/min using acetonitrile - a solution containing 10mM ammonium bicarbonate at suitable ratio. The ion transitions at m/z 605.4→ 249.1 (leuprolide) and m/z 152 → 110 (paracetamol) were monitored under the positive ion mode of electrospray ionization with multiple reaction monitoring. The standard curves were linear in the range of 0.05 to 50 ng/ml for Leuprolide with a mean correlation coe¶cient greater than 0.99. The intra-day and inter-day precision and accuracy were within 93.9% - 119.1% with CV≤ 10%. The assay was applied to determine concentrations of leuprolide in dog serum in pharmacokinetic and bioavailability studies.