Pharmacokinetics and Metabolism, Drug Safety and Pharmacokinetics Laboratories, Taisho Pharmaceutical Co., Ltd., 1-403 Yoshino-cho, Kita-ku, Saitama-shi, Saitama 331-9530, Japan.
J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Feb 15;885-886:8-14. doi: 10.1016/j.jchromb.2011.11.037. Epub 2011 Dec 13.
A highly selective, sensitive, and robust liquid chromatography-tandem mass spectrometric method for the determination of 5-thio-d-glucose concentrations in rat and human plasma was developed and validated. The sample preparation procedure involved protein precipitation and solid phase extraction, which efficiently removed sources of interference present in the plasma. Chromatographic separation was obtained using an NH(2)-column with distilled water and acetonitrile as the mobile phase under gradient conditions. Detection was performed using tandem mass spectrometry equipped with an electrospray ionization interface in negative ion mode. The selected reaction monitoring (SRM) transitions for 5-thio-d-glucose and an internal standard (5-thio-d-glucose-(13)C(6)) were m/z 195→m/z 105 and m/z 201→m/z 108, respectively. The correlation coefficients of the calibration curves ranged from 0.9997 to 0.9999 over a concentration range from 10 to 3000ng/mL plasma. The validated method was successfully applied to a pharmacokinetic study in rats.
建立并验证了一种用于测定大鼠和人血浆中 5-硫代-d-葡萄糖浓度的高选择性、高灵敏度和稳健的液相色谱-串联质谱法。样品前处理步骤包括蛋白沉淀和固相萃取，可有效去除血浆中存在的干扰源。采用 NH(2)柱，以蒸馏水和乙腈为流动相，在梯度条件下进行色谱分离。检测采用电喷雾电离接口在负离子模式下进行串联质谱分析。5-硫代-d-葡萄糖和内标（5-硫代-d-葡萄糖-（13）C（6））的选择反应监测（SRM）跃迁分别为 m/z 195→m/z 105 和 m/z 201→m/z 108。校准曲线的相关系数在 10 至 3000ng/mL 血浆浓度范围内为 0.9997 至 0.9999。该验证方法成功应用于大鼠的药代动力学研究。
