超高效液相色谱-串联质谱法研究亚硫酸氢钠穿心莲内酯在大鼠体内排泄  

UHPLC-tandem mass spectrometry investigation of andrographolide sodium bisulphite excretion in rats

作　　者：张双庆[1] 孙旭[1] 于敏[1] 李佐刚[1] 

机构地区：[1]中国食品药品检定研究院,食品药品安全评价研究所,药物代谢动力学实验室,北京100176

出　　处：《药物分析杂志》2012年第10期1736-1740,共5页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立超高效液相色谱-串联质谱法测定大鼠尿液和粪便中亚硫酸氢钠穿心莲内酯(ASB)含量,并用于研究大鼠静脉注射ASB在尿液和粪便中的排泄。方法:采用甲醇提取生物样品中的ASB,以脱水穿心莲内酯(DAG)为内标,在HypersilGold C18色谱柱(50 mm×2.1 mm,1.9μm)上以流动相甲醇-水梯度洗脱,流速0.2 mL·min-1,柱温35℃,分析时间6 min;在电喷雾离子化电离源上以选择反应监测方式进行负离子检测,用于定量分析的离子反应分别为m/z 413.2→287.2(ASB)和m/z 331.2→303.3(DAG)。大鼠自尾静脉注射80 mg.kg-1ASB,收集并测定给药后各时段的尿液和粪便中ASB浓度,计算各时段尿液和粪便中累计排泄率。结果:测定ASB的线性范围为50～5000 ng·mL-1,方法定量下限为50 ng·mL-1。日内和日间精密度(RSD)分别在11.2%和13.3%以内,准确度分别在85.8%～101.4%和87.9%～97.5%,提取回收率为96.1%～98.3%,基质效应为96.2%～98.1%。大鼠72 h尿液中累计排泄率为16.9%±3.9%,粪便中为34.1%±18.4%。结论:本方法速度快、专属性好、准确度高,适用于大鼠尿液和粪便中ASB含量测定,可应用于ASB大鼠排泄研究。Objective：To establish an ultra-high performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS） method for quantification of andrographolide sodium bisulphite（ASB） in rat urine and feces,and investigate excretion of ASB in rats after intravenous administration.Methods：ASB was extracted from biological samples using methanol and separated on a Hypersil Gold C18 column（50 mm× 2.1 mm,1.9 μm）with gradient mobile phase consisting of methanol and water at a flow rate of 0.2 mL·min-1.Dehydroandrographolide（DAG） was adopted as an internal standard.Under negative electrospray ionization mode,reaction monitoring of the precursor-product ion transitions m/z 413.2→287.2 was selected for ASB and m/z 331.2→303.3 for DAG.Urine and feces were collected at different time points after intravenously admonition of with ASB at a single dose of 80 mg·kg-1.Results：The linear calibration curve was obtained in the concentration range of 50-5000 ng·mL-1 with a minimal quantification limit of 50 ng·mL-1.Intra-and inter-day precision was less than 11.2% and 13.3%,respectively.Intra-and inter-day accuracy fell in the ranges of 85.8%-101.4% and 87.9%-97.5%,respectively.The recovery ranged from 96.1% to 98.3%,and the matrix effects were between 96.2% and 98.1%.The cumulative amounts of ASB excreted in urine and feces within 72 h as parent drug were 16.9%±3.9% and 34.1%±18.4% of administered dose,respectively.Conclusion：The method is rapid,specific and accurate,which is suitable for ASB quantification in urine and feces and can be applied to ASB excretion study in rats.

关 键 词：亚硫酸氢钠穿心莲内酯 超高效液相色谱-串联质谱 尿液与粪便样品 安全性评价 感染性疾病 注射剂 排泄 

分 类 号：R917[医药卫生—药物分析学]