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不同种属血浆中的稳定性  

UPLC-MS/MS investigation of stability of andrographolide sodium bisulphite in different species plasma

作　　者：于敏[1] 张双庆[1] 孙旭[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12期2144-2149,共6页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立超高效液相色谱-串联质谱的方法,测定血浆中亚硫酸氢钠穿心莲内酯(ASB)的含量,并用于研究ASB在不同种属血浆中的稳定性。方法:ASB血浆样品用甲醇进行提取,以脱水穿心莲内酯(DAG)为内标进行定量。采用HypersilGold C18(50 mm×2.1 mm,1.9μm)色谱柱,柱温35℃;流动相为水-甲醇梯度洗脱,流速0.2 mL.min-1;分析时间6 min。采用负离子电喷雾离子化电离源和选择反应监测(SRM)模式,进行串联质谱分析;用于定量分析的离子反应分别为m/z413.2→287.2(ASB)和m/z 331.2→303.3(DAG)。浓度为1μg.mL-1的ASB大鼠、犬、人血浆液37℃孵育,于不同时间点测定ASB含量,计算血浆中ASB剩余百分含量。结果:ASB标准血浆样品的线性范围在10～1000 ng.mL-1,定量下限为10 ng.mL-1。方法的日内和日间准确度分别在96.4%～103.2%和93.7%～98.0%,精密度(RSD)分别小于6.9%和10.8%,基质效应为88.9%～99.9%,提取回收率为71.2%～98.9%。大鼠、犬和人血浆中ASB剩余百分含量均明显下降,在犬血浆中5 min血药浓度降至70%的较高水平,在大鼠和人血浆中5 min降至30%左右,之后趋于平稳。结论:本方法适用于血浆中ASB的含量测定,ASB在不同种属血浆中下降均很快,但表现出明显的种属差异。Objectives：To establish a high performance liquid chromatography -tandem mass spectrometry （HPLC -MS/MS） method for the determination of andrographolide sodium bisulphite （ASB） in plasma, and adopt the method to evaluate stability of ASB in plasma of different species. Methods：The analyte from plasma samples ex- tracted by methanol and dehydroandrographolide （DAG） was used as an internal standard for quantitation. The chromatographic separation was achieved within 6 min on a Hypersil Gold C18 column with gradient mobile phase containing a mixture of water and methanol at a flow rate of 0.2 mL · min-1. Column temperature was 35 R2. At negative electrospray ionization mode, selected reaction monitoring of the precursor - product ion transitions of m/z 413.2--＋287.2 for ASB and m/z 331.2-303.3 for DAG was used for quantification. The remaining amount of ASB incubated in rat, dog and human plasma was determined at different time points. Results：The calibration curves were linear in the range of 10 - 1000 ng · mL-1 with a lower detection limit of 10 ng · mL-1. Intra - and inter - day accuracy fell in the ranges of 96.4% - 103.2% and 93.7% - 98.0% , respectively. Intra - and inter - clay precision was less than 6.9% and 10. 8% ,respectively. The matrix effects ranged from 88.9% to 99.9% and the recovery was between 71.2% and 98.9%. The remaining percentage content of ASB in three species plasma was obviously declined. The ASB remaining content incubated 5 minutes in dog plasma was down to 70% , and in rats and human plasma were down to 30%. Conclusion：The method was suitable for quantification of ASB in plasma. Itwas showed that ASB remaining content decreased rapidly in plasma of rat, dog and human. But there was differ- ence in species.

关 键 词：亚硫酸氢钠穿心莲内酯 脱水穿心莲内酯 高效液相色谱-串联质谱 方法验证 药物血浆稳定性研究 

分 类 号：R917[医药卫生—药物分析学]