Wang Bao-Lian, Hu Jin-Ping, Tan Wei, Sheng Li, Chen Hui, Li Yan
Institute of Materia Medica, Chinese Academy of Medical Sciences & Perking Union Medical College, Beijing 100050, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2008 Apr 1;865(1-2):114-20. doi: 10.1016/j.jchromb.2008.02.016. Epub 2008 Mar 4.
A simple, rapid and sensitive method was developed for the simultaneous quantification of four active schisandra lignans (schisandrin, schisantherin A, deoxyshisandrin and gamma-schisandrin) from a traditional Chinese medicine Schisandra chinensis(Wuweizi) in rat plasma using a high-performance liquid chromatography system coupled to a positive ion electrospray mass spectrometric analysis. The plasma sample preparation was a simple deproteinization by the addition of three volumes of methanol followed by centrifugation. The analytes and internal standard (IS) bicyclol were separated on a Zorbax SB-C18 column (3.5 microm, 2.1 mm x 100 mm) with mobile phase of methanol/water (70:30, v/v) containing 0.1% formic acid at a flow rate of 0.2 mL/min with an operating temperature of 25 degrees C. Detection was performed on a Trap XCT mass spectrometer equipped with an electrospray ionization (ESI) source operated in selected ion monitoring (SIM) mode. Positive ion ESI was used to form sodium adduct molecular ions at m/z 455 for schisandrin, m/z 559 for schisantherin A, m/z 439 for deoxyshisandrin, m/z 423 for gamma-schisandrin, and m/z 413 for the internal standard bicyclol. Linear detection responses were obtained for the four test compounds ranging from 0.010 to 2.0 microg/mL and the lower limits of quantitation (LLOQs) for four lignans were 0.010 microg/mL. The intra- and inter-day precisions (R.S.D.%) were within 12.5% for all analytes, while the deviation of assay accuracies was within +/-13.0%. The average recoveries of analytes were greater than 80.0%. All analytes were proved to be stable during all sample storage, preparation and analytic procedures. The method was successfully applied to the pharmacokinetic study of the four lignans after oral administration of Schisandra chinensis extraction to rats.
建立了一种简单、快速且灵敏的方法,用于同时定量大鼠血浆中四种活性五味子木脂素(五味子醇甲、五味子酯甲、脱氧五味子醇甲和γ-五味子醇甲),该方法采用高效液相色谱系统与正离子电喷雾质谱分析联用,从传统中药五味子(五味子)中提取。血浆样品制备方法简单,加入三倍体积的甲醇进行脱蛋白,然后离心。分析物和内标(IS)双环醇在Zorbax SB-C18柱(3.5微米,2.1毫米×100毫米)上分离,流动相为甲醇/水(70:30,v/v),含0.1%甲酸,流速为0.2毫升/分钟,操作温度为25℃。在配备电喷雾电离(ESI)源的Trap XCT质谱仪上进行检测,采用选择离子监测(SIM)模式。正离子ESI用于形成五味子醇甲的m/z 455、五味子酯甲的m/z 559、脱氧五味子醇甲的m/z 439、γ-五味子醇甲的m/z 423以及内标双环醇的m/z 413的钠加合物分子离子。四种测试化合物的线性检测响应范围为0.010至2.0微克/毫升,四种木脂素的定量下限(LLOQs)为0.010微克/毫升。所有分析物的日内和日间精密度(R.S.D.%)均在12.5%以内,测定准确度偏差在±13.0%以内。分析物的平均回收率大于80.0%。所有分析物在所有样品储存、制备和分析过程中均被证明是稳定的。该方法成功应用于大鼠口服五味子提取物后四种木脂素的药代动力学研究。