Sheng Ning, Yuan Lin, Zhi Xuran, Cui Cheng, Zhang Zhiyong, Jia Peipei, Zhang Xiaoxu, Zhang Lantong, Wang Xinguo
Department of Pharmaceutical Analysis, School of Pharmacy, Hebei Medical University, 361 East Zhongshan Road, Shijiazhuang, Hebei 050017, PR China; Department of Pharmacy, The Third Hospital of hebei Medical University, 139 Ziqiang Road, Shijiazhuang, Hebei 050051, PR China; Department of Traditional Chinese Medicine Pharmacology, School of Traditional Chinese Medicine, Hebei Medical University, 326 Xinshinan Road, Shijiazhuang, Hebei 050091, PR China.
Department of Pharmaceutical Analysis, School of Pharmacy, Hebei Medical University, 361 East Zhongshan Road, Shijiazhuang, Hebei 050017, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Oct 15;969:1-11. doi: 10.1016/j.jchromb.2014.07.042. Epub 2014 Aug 7.
A sensitive, reliable and accurate high-performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS) was developed and validated for the quantification of sweroside in rat plasma, tissue and excretion. A single-step protein precipitation by methanol was used to prepare samples. Sweroside and swertiamarin (internal standard, IS) were separated by using a C18 column and a mobile phase consisted of methanol and water containing 0.1% formic acid running at a flow rate of 0.8ml/min for 6min. Detection and quantification were performed using a mass spectrometer by the multiple-reaction monitoring (MRM) in positive electrospray ionization mode. The optimized mass transition ion pairs (m/z) for quantitation were M+H359.1→197.2 for sweroside and M+Na397.4→165.3 for swertiamarin (IS), respectively. The inter-day precision (RSD %) was less than 11.20% and intra-day precision (RSD %) was less than 10.90%, while the inter-day accuracy (RE %) was ranged from -9.69 to 9.17% and intra-day accuracy (RE %) was ranged from -10.56 to 13.47%. The mean elimination half-life (t1/2) of sweroside for 5, 10 and 15mg/kg dose were 78.8, 67.6 and 77.2min, respectively. And sweroside follows linear plasma pharmacokinetics across the investigated dosage range in rats (5-15mg/kg). The absolute bioavailability (F %) of sweroside was 11.90% on average. The results of tissue distribution showed the higher sweroside concentrations were found in kidney, liver, spleen and lung, and the small amount of drug was distributed into the brain tissue. The high distribution in liver confirms the reports that sweroside has hepatoprotective activity and promoted liver regeneration, and there was no long-term accumulation of sweroside in rat tissues. Total recoveries of sweroside within 48h were 0.67% in bile, 1.55% in urine and 0.46% in feces, which might be resulted from liver first-pass effect. The above results suggested that sweroside was mainly excreted as the metabolites.
建立了一种灵敏、可靠且准确的高效液相色谱-串联质谱法（HPLC-MS/MS），并对其进行验证，用于定量大鼠血浆、组织及排泄物中的獐牙菜苷。采用甲醇一步沉淀蛋白法制备样品。使用C18色谱柱分离獐牙菜苷和獐牙菜苦苷（内标，IS），流动相由甲醇和含0.1%甲酸的水组成，流速为0.8ml/min，运行6min。采用质谱仪在正电喷雾电离模式下通过多反应监测（MRM）进行检测和定量。定量的优化质量转移离子对（m/z）分别为獐牙菜苷的M+H359.1→197.2和獐牙菜苦苷（IS）的M+Na397.4→165.3。日间精密度（RSD%）小于11.20%，日内精密度（RSD%）小于10.90%，而日间准确度（RE%）范围为-9.69至9.17%且日内准确度（RE%）范围为-10.56至13.47%。5、10和15mg/kg剂量的獐牙菜苷平均消除半衰期（t1/2）分别为78.8、67.6和77.2min。并且在研究的大鼠剂量范围（5-15mg/kg）内，獐牙菜苷呈现线性血浆药代动力学特征。獐牙菜苷的绝对生物利用度（F%）平均为11.90%。组织分布结果显示，在肾脏、肝脏、脾脏和肺中发现獐牙菜苷浓度较高，且少量药物分布到脑组织中。在肝脏中的高分布证实了獐牙菜苷具有肝保护活性并促进肝脏再生的报道，并且獐牙菜苷在大鼠组织中没有长期蓄积。48h内獐牙菜苷在胆汁中的总回收率为0.67%，在尿液中为1.55%，在粪便中为0.46%，这可能是由于肝脏首过效应所致。上述结果表明獐牙菜苷主要以代谢产物形式排泄。
