Song Min, Hang Tai-Jun, Wang Cheng, Yang Lin, Wen Ai-Dong
Department of Pharmaceutical Analysis, China Pharmaceutical University, Nanjing 210009, China.
Department of Pharmacy, Xijing Hospital, Xi'an 710032, China.
J Pharm Anal. 2012 Feb;2(1):19-28. doi: 10.1016/j.jpha.2011.08.003. Epub 2011 Oct 24.
A selective precolumn derivatization liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the determination of glucosamine in human plasma and urine has been developed and validated. Glucosamine was derivatized by o-phthalaldehyde/3-mercaptopropionic acid. Chromatographic separation was performed on a Phenomenex ODS column (150 mm×4.6 mm, 5 μm) using linear gradient elution by a mobile phase consisting of methanol (A), and an aqueous solution containing 0.2% ammonium acetate and 0.1% formic acid (B) at a flow rate of 1 mL/min. Tolterodine tartrate was used as the internal standard (IS). With protein precipitation by acetonitrile and then the simple one-step derivatization, a sensitive bio-assay was achieved with the lower limit of quantitation (LLOQ) as low as 12 ng/mL for plasma. The standard addition calibration curves suitable for clinical sample analysis showed good linearity over the range of 0.012-8.27 μg/mL in plasma and 1.80-84.1 μg/mL in urine. The fully validated method has been successfully applied to a pharmacokinetic study of compound glucosamine sulfate dispersible tablets in health Chinese volunteers receiving single oral doses at 500, 1000 and 1500 mg of glucosamine sulfate, as well as multiple oral doses of 500 mg t.i.d. for 7 consecutive days.
已开发并验证了一种用于测定人血浆和尿液中氨基葡萄糖的选择性柱前衍生化液相色谱 - 串联质谱（LC-MS/MS）方法。氨基葡萄糖用邻苯二甲醛/3-巯基丙酸进行衍生化。色谱分离在Phenomenex ODS柱（150 mm×4.6 mm，5μm）上进行，使用由甲醇（A）和含有0.2%醋酸铵和0.1%甲酸的水溶液（B）组成的流动相进行线性梯度洗脱，流速为1 mL/min。酒石酸托特罗定用作内标（IS）。通过乙腈沉淀蛋白质，然后进行简单的一步衍生化，实现了一种灵敏的生物测定方法，血浆定量下限（LLOQ）低至12 ng/mL。适用于临床样品分析的标准加入校准曲线在血浆中0.012 - 8.27μg/mL和尿液中1.80 - 84.1μg/mL范围内显示出良好的线性。该经过充分验证的方法已成功应用于健康中国志愿者单次口服500、1000和1500 mg硫酸氨基葡萄糖以及连续7天每天3次口服500 mg硫酸氨基葡萄糖复方硫酸氨基葡萄糖分散片的药代动力学研究。
