Key Laboratory of Drug Quality Control and Pharmacovigilance, Ministry of Education, China Pharmaceutical University, Nanjing 210009, China.
J Anal Toxicol. 2009 Nov-Dec;33(9):588-94. doi: 10.1093/jat/33.9.588.
A rapid, sensitive, and specific liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS-MS) method was developed and validated for simultaneous determination of aconitine (AC), mesaconitine (MA), and hypaconitine (HA), the three toxic constituents from Sini decoction (SND) in rat plasma. After the addition of citalopram as the internal standard (IS), plasma samples were basified with 100 microL 10% ammonium hydroxide, and then extracted with 1 mL ethyl acetate. Chromatographic separation was performed on a CN column (250 mm x 4.6 mm, 5 microm) with a mobile phase of methanol/40 mM ammonium acetate/formic acid (950:45:5, v/v/v) at the flow rate of 1.0 mL/min. Analytes were determined in a triple-quadrupole mass spectrometer in the selected reaction-monitoring (SRM) mode using electrospray source with positive mode. The method was validated over the concentration ranges of 0.01-10 ng/mL for AC, MA, and HA. The variation coefficients were always < 15% for both intraday and interday precision for each analyte. Mean accuracies were also within +/-15%. The method was proved to be sensitive, rapid, specific, accurate, and reproducible. It has been successfully applied to the pharmacokinetics study on rats after oral administration of SND.
建立并验证了一种灵敏、特异的液相色谱-电喷雾串联质谱（LC-ESI-MS-MS）方法，用于同时测定大鼠血浆中的乌头碱（AC）、次乌头碱（MA）和新乌头碱（HA）这三种来自四逆汤（SND）的有毒成分。在加入西酞普兰作为内标（IS）后，将血浆样品用 100μL10%氨水溶液碱化，然后用 1mL 乙酸乙酯提取。采用 CN 柱（250mm×4.6mm，5μm），以甲醇/40mM 乙酸铵/甲酸（950:45:5，v/v/v）为流动相，流速为 1.0mL/min 进行色谱分离。在电喷雾源正模式下，采用选择反应监测（SRM）模式，在三重四极杆质谱仪上进行分析物的测定。该方法在 AC、MA 和 HA 的浓度范围为 0.01-10ng/mL 时得到验证。每个分析物的日内和日间精密度的变异系数始终<15%。平均准确度也在±15%范围内。该方法灵敏、快速、特异、准确、重现性好。已成功应用于 SND 灌胃后大鼠的药代动力学研究。
