HPLC法测定大鼠肝微粒体中6-羟基氯唑沙宗的含量
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摘要
目的:建立高效液相色谱法测定大鼠肝微粒体中6-羟基氟唑沙宗的含量。方法:采用Diamonsil C_(18)柱(200 mm×4.6 mm,5μm),以甲醇-水-磷酸(45:55:0.1)为流动相,流速1.0 ml·min~(-1),检测波长为296 nm。结果:6-羟基氯唑沙宗浓度在0.5~25μmol·L~(-1)范围内呈现良好的线性关系(r=0.999 1),方法回收率为92.57%~98.72%,提取回收率为76.38%~82.25%,日内、日间精密度RSD<9%。结论:该方法可靠、准确、重复性好,可用于大鼠肝微粒体中6-羟基氯唑沙宗的含量测定及体外代谢研究。
Objective:To establish an HPLC method for the determination of 6-hydroxy chlorzoxazone in rat liver microsomes.Methods:The samples were separated on a RP-C_(18) column(Diamonsil,200 mm×4.6 mm,5 μm) with methanol-water-phosphoric acid(45:55:0.1) as the mobile phase at a flow rate of 1.0 ml·min~(-1) and the detection wavelength was at 296 nm.Results:The calibration curve was linear over the range of 0.5-25 μmol·L~(-1) for 6-hydroxy chlorzoxazone(r=0.999 1).The relative recovery of the samples ranged from 92.57%-98.72%.The absolute recovery of the samples ranged from 76.38%-82.25%.The intra and interday precision RSD were below 9%.Conclusion:The method is reliable,precise and reproducible in the content determination of 6-hydroxy chlorzoxazone in rat liver microsomes and metabolism ia vitro.
Objective:To establish an HPLC method for the determination of 6-hydroxy chlorzoxazone in rat liver microsomes.Methods:The samples were separated on a RP-C_(18) column(Diamonsil,200 mm×4.6 mm,5 μm) with methanol-water-phosphoric acid(45:55:0.1) as the mobile phase at a flow rate of 1.0 ml·min~(-1) and the detection wavelength was at 296 nm.Results:The calibration curve was linear over the range of 0.5-25 μmol·L~(-1) for 6-hydroxy chlorzoxazone(r=0.999 1).The relative recovery of the samples ranged from 92.57%-98.72%.The absolute recovery of the samples ranged from 76.38%-82.25%.The intra and interday precision RSD were below 9%.Conclusion:The method is reliable,precise and reproducible in the content determination of 6-hydroxy chlorzoxazone in rat liver microsomes and metabolism ia vitro.
引文
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2伍忠奎,谢红光,周宏灏.细胞色素P4502EI的研究进展[J].中国临床药理学杂志,1997,13(1):57-62
3秦梦楠,刘蕊,刘高峰,等.灯盏花素注射液对大鼠体外肝微粒体细胞色素P450酶活性的影响[J].中国药师,2012,15(2):147-150
4 Bradford MM.A rapid anti sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J].Analytical Biocbemistry,1976,72:248-254
5闵鹏,闻晶,赵庆春,等.胡柚提取物抑制硝苯地平肝微粒体酶代谢的研究[J].中因药师.2012,15(12):1693-1695
6 Lu Y,Cederbaum AI.CYP2EI and oxidative liver injury by alcohol[J].Free radical biology&medicine,2008,44(5):723-738
7夏军,樊宝才,谢文明,等.雷公藤多苷片对大鼠药物代谢酶CYP2E1和CYP3A4活性的影响[J].中国药师,2012,15(11):1529-1532
8孙鲁宁,丁黎,严拯字,等.LC-MS/MS法研究苯环喹溴铵对大鼠肝微粒体CYP450酶的抑制作用[J].沈阳药科大学学报,2013,44(2):134-140
2伍忠奎,谢红光,周宏灏.细胞色素P4502EI的研究进展[J].中国临床药理学杂志,1997,13(1):57-62
3秦梦楠,刘蕊,刘高峰,等.灯盏花素注射液对大鼠体外肝微粒体细胞色素P450酶活性的影响[J].中国药师,2012,15(2):147-150
4 Bradford MM.A rapid anti sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J].Analytical Biocbemistry,1976,72:248-254
5闵鹏,闻晶,赵庆春,等.胡柚提取物抑制硝苯地平肝微粒体酶代谢的研究[J].中因药师.2012,15(12):1693-1695
6 Lu Y,Cederbaum AI.CYP2EI and oxidative liver injury by alcohol[J].Free radical biology&medicine,2008,44(5):723-738
7夏军,樊宝才,谢文明,等.雷公藤多苷片对大鼠药物代谢酶CYP2E1和CYP3A4活性的影响[J].中国药师,2012,15(11):1529-1532
8孙鲁宁,丁黎,严拯字,等.LC-MS/MS法研究苯环喹溴铵对大鼠肝微粒体CYP450酶的抑制作用[J].沈阳药科大学学报,2013,44(2):134-140