Department of Physiological Sciences, Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, OK 74078, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2013 Jan 15;913-914:147-54. doi: 10.1016/j.jchromb.2012.11.012. Epub 2012 Dec 20.
A highly sensitive and specific liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry (LC/APCI-MS/MS) method has been developed and validated for simultaneous quantification of vinblastine and its metabolite, desacetylvinblastine, in canine plasma and urine samples. Plasma and urine samples were processed by a solid phase extraction procedure. The optimal chromatographic behavior of these analytes was achieved on pentafluorophenyl (PFP) propyl analytical column (5μm, 50×2.1mm) under isocratic elution of 0.75mL/min with a mobile phase of 5mM ammonium acetate and methanol. The samples were analyzed in positive ion, multiple reaction monitoring mode. The calibration curves were linear over 0.125-2ng/mL (lower calibration curve); 2-100ng/mL (higher calibration curve) and 0.125-5ng/mL for vinblastine and desacetylvinblastine in plasma, and over 1-2000ng/mL and 0.5-100ng/mL for vinblastine and desacetylvinblastine in urine samples, respectively. The limits of quantitation of vinblastine and desacetylvinblastine were 0.125ng/mL in both matrices. The intra and interday accuracy was above 89% and precision below 8.6% for both analytes in both matrices. The developed method was successfully applied to ongoing in vivo vinblastine pharmacokinetic studies in dogs.
建立并验证了一种灵敏、特异的液相色谱-大气压化学电离-质谱联用（LC/APCI-MS/MS）法，用于犬血浆和尿液样品中长春碱及其代谢物去乙酰长春碱的同时定量分析。血浆和尿液样品经固相萃取处理。这些分析物在五氟苯基（PFP）丙基分析柱（5μm，50×2.1mm）上以 0.75mL/min 的等度洗脱、流动相为 5mM 乙酸铵和甲醇的条件下，可实现最佳的色谱行为。采用正离子、多反应监测模式进行分析。长春碱和去乙酰长春碱在血浆中的校准曲线在 0.125-2ng/mL（下校准曲线）；2-100ng/mL（高校准曲线）和 0.125-5ng/mL 范围内呈线性，在尿液中的校准曲线在 1-2000ng/mL 和 0.5-100ng/mL 范围内呈线性。长春碱和去乙酰长春碱在两种基质中的定量下限均为 0.125ng/mL。在两种基质中，长春碱和去乙酰长春碱的日内和日间精密度均在 8.6%以下，准确度均在 89%以上。该方法成功应用于犬体内长春碱药代动力学研究。
