杜玥, 陈笑艳, 杨汉煜, 钟大放. 液相色谱-串联质谱法测定大鼠血浆中的汉黄芩素J. 药学学报, 2002, 37(5): 362-366.
引用本文: 杜玥, 陈笑艳, 杨汉煜, 钟大放. 液相色谱-串联质谱法测定大鼠血浆中的汉黄芩素J. 药学学报, 2002, 37(5): 362-366.
DU Yue, CHEN Xiao-yan, YANG Han-yu, ZHONG Da-fang. DETERMINATION OF WOGONIN IN RAT PLASMA BY LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRYJ. Acta Pharmaceutica Sinica, 2002, 37(5): 362-366.
Citation: DU Yue, CHEN Xiao-yan, YANG Han-yu, ZHONG Da-fang. DETERMINATION OF WOGONIN IN RAT PLASMA BY LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRYJ. Acta Pharmaceutica Sinica, 2002, 37(5): 362-366.

液相色谱-串联质谱法测定大鼠血浆中的汉黄芩素

DETERMINATION OF WOGONIN IN RAT PLASMA BY LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY

  • 摘要: 目的建立测定大鼠血浆中汉黄芩素浓度的液相色谱-串联质谱法。方法血浆样品经液-液萃取后,进行色谱分离,采用三重四极杆串联质谱检测,使用大气压化学电离源(APCI),在正离子条件下选择反应监测(SRM)方式进行扫描,用于定量分析的离子分别为m/z 284.8→m/z 269.5(汉黄芩素)和m/z 254.7→m/z 198.5(葛根黄豆苷元,内标)。结果线性范围为0.25~20 ng·mL-1,最低定量浓度为0.25 ng·mL-1。以质控样品(QC)计算,在各浓度水平下,此法的批内精密度(RSD)为2.2%~13.1%,批间精密度为5.9%~7.3%;准确度(RE)为-0.3%~1.3%。结论 此法灵敏、快速、准确,可用于大鼠血浆中汉黄芩素浓度测定及临床前药动学研究。

     

    Abstract: AIMTo develop a sensitive and rapid LC/MS/MS method for the determination of wogonin, an active flavonoid shown to have an inhibitory effect on the proliferation of a carcinoma cell line, in rat plasma after an oral administration. METHODSWogonin and daidzein (internal standard) were extracted from plasma directly with n-hexane-diethyl ether (1∶4). After liquid-liquid extraction, the analytes of interest were separated on a Diamonsil C18 column. The mobile phase was acetonitrile-water-formic acid (80∶20∶1) with a flow rate of 0.8 mL·min-1. A Finnigan TSQ (triple stage quadruple) tandem mass spectrometer equipped with an atmospheric pressure chemical ionization (APCI) source was used as detector and was operated in positive ion mode. Selected reaction monitoring (SRM) was used and transitions selected for quantitation were: m/z 284.8→269.5 for wogonin and m/z 254.7→198.5 for daidzein. The mass spectrometric conditions were as follows: The temperatures of the vaporizer and heated capillary were 450℃ and 250℃, respectively. The corona discharge current was 4.00 μA. Nitrogen was used as the sheath and auxiliary gas, whose settings were 0.6 MPa and 3 mL·min-1, respectively. Argon was used as the collision gas at a pressure of 1.4 Pa. The collision energy of 35 V was chosen for both wogonin and daidzein. RESULTSThe calibration curve was linear over the concentration range of 0.25 to 20.0 ng·mL-1. The limit of quantitation was 0.25 ng·mL-1. Within-day and between-day precision expressed by relative standard deviation (RSD) was 2.2% to 13.1% and 5.9% to 7.3%, respectively, and the accuracy expressed by RE was -0.3% to 1.3%. CONCLUSIONThis method proved to be specific, accurate and sensitive enough to be applied to the pharmacokinetic studies of wogonin in rats after a single dose of 5 mg·kg-1 by oral administration.

     

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