Zhu Hong, Ren Yanping, Sun Yingguang, Chang Lu, Cao Liang, Xu Huijin, Zhang Lantong
Department of Pharmaceutical Analysis, School of Pharmacy, Hebei Medical University, Shijiazhuang, China.
Biomed Chromatogr. 2013 Apr;27(4):527-34. doi: 10.1002/bmc.2827. Epub 2012 Sep 27.
A simple, fast and sensitive method for the simultaneous determination of cnidilin and its two metabolites (M1 and M2) in rat bile and stool using HPLC coupled with electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) has been developed. The sample pretreatment was simple, because methanol was the only additive used for dilution of bile and ultrasound of stool. Pimpinellin was used as internal standard (IS). The separation was performed on a reverse phase C18 column with gradient elution consisting of 0.5‰ aqueous formic acid and methanol (containing 0.5‰ formic acid). The detection was in the multiple-reaction monitoring mode within 7 min. All the analytes were in accordance with the requirement of the validation of the method in vivo (linearity, precision, accuracy, limit of detection and limit of quantification). After oral administrating 24 mg/kg of the prototype drug cnidilin, M1 and M2 were determined in bile within 36 h, and in stool within 60 h. Cnidilin in bile was completely excreted in 24 h, and the main excretive amount of cnidilin was 80% in the first 6 h, but the drug recovery in bile within 24 h was <1.95%. In stool, the main excretive amount of cnidilin was 95.8% in the first 24 h, and the drug recovery within 48 h was lower than 1.48%.
已开发出一种简单、快速且灵敏的方法，用于使用高效液相色谱-电喷雾电离串联质谱法（HPLC-ESI-MS/MS）同时测定大鼠胆汁和粪便中的蛇床子素及其两种代谢物（M1和M2）。样品预处理简单，因为甲醇是用于稀释胆汁和对粪便进行超声处理的唯一添加剂。茴芹内酯用作内标（IS）。在反相C18柱上进行分离，采用由0.5‰甲酸水溶液和甲醇（含0.5‰甲酸）组成的梯度洗脱。在7分钟内以多反应监测模式进行检测。所有分析物均符合体内方法验证的要求（线性、精密度、准确度、检测限和定量限）。口服给予24 mg/kg的原型药物蛇床子素后，在36小时内在胆汁中检测到M1和M2，在60小时内在粪便中检测到。胆汁中的蛇床子素在24小时内完全排泄，蛇床子素的主要排泄量在前6小时为80%，但24小时内胆汁中的药物回收率<1.95%。在粪便中，蛇床子素的主要排泄量在前24小时为95.8%，48小时内的药物回收率低于1.48%。
