摘要
目的建立LC-MS法同时测定大鼠血浆中熊去氧胆酸和鹅去氧胆酸的含量,并研究灌胃给药熊胆粉提取物后熊去氧胆酸和鹅去氧胆酸在大鼠体内的药物代谢动力学行为。方法血浆样品采用甲醇、乙酸乙酯和甲酸（50∶50∶1）混合溶剂处理,以双氯芬酸作为内标,色谱柱为Waters Atlantis T3 C18色谱柱（150 mm×2.0 mm,5μm）,流动相为水（含1 mmol·L-1醋酸铵和0.01%甲酸）-甲醇（76∶24）,等度洗脱,流速为0.2 m L·min-1,岛津LC-MS2020质谱单四极杆检测器,负离子模式。大鼠灌胃100 mg·kg-1的熊胆粉提取物,LC-MS法测定熊去氧胆酸和鹅去氧胆酸的血药浓度,并采用DAS 2.0软件计算药物代谢动力学参数。结果方法学实验结果表明内源性杂质不干扰熊去氧胆酸、鹅去氧胆酸和内标的测定,线性范围0.078~10.0μg·m L-1,定量限为0.078μg·m L-1。方法精密度、准确度、稳定性和回收率均符合生物样品测定的要求,适合大鼠血浆中2种胆酸浓度的测定,可以应用该方法进行熊去氧胆酸和鹅去氧胆酸的药物代谢动力学研究。结论本方法操作简便、灵敏、专属性强,方法学考证符合生物样品测定的要求并成功用于熊去氧胆酸和鹅去氧胆酸在大鼠体内的药物代谢动力学研究。
Objective To establish an LC-MS method to determine ursodeoxycholic acid and chenodeoxycholic acid in rat plasma and to investigate pharmacokinetic properties of ursodeoxycholic acid and chenodeoxycholic acid after an oral administration of bear gall powder extract to rats. Methods Ursodeoxycholic acid and chenodeoxycholic acid and diclofenac（the internal standard, IS） were extracted by liquid-liquid extraction and separated on a Waters Atlantis T3 C18 column, with methanol and water containing 1 mmol·L- 1 ammonium acetate and 0.01% formic acid（24：76） as the mobile phase. Quantitation was carried out by single reaction monitoring in negative mode on a Shimadzu LC-MS2020 system. After the validation, the developed method was used to evaluate the pharmacokinetics of ursodeoxycholic acid and chenodeoxycholic acid in rats after an oral administration of 100 mg·kg- 1 of bear gall powder extract. Results The study showed a good linearity at 0.078-10.0 μg·m L- 1 with a sensitivity of 0.078 μg·m L- 1 as the limit of quantification. The precision, accuracy, stability and mean recoveries were satisfied and met the requirement of biological sample measurement. The method was successfully applied in the pharmacokinetic study of ursodeoxycholic acid and chenodeoxycholic acid in rat plasma samples. Conclusion This LC-MS method is selective, sensitive and reproducibile, and is successfully applied in the a pharmacokinetic study of ursodeoxycholic acid and chenodeoxycholic acid in rats.
出处
《中南药学》
CAS
2016年第7期716-720,共5页
Central South Pharmacy
基金
盐城市科技项目医药类资助(No.20143115)
