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作 者:刘佳[1] 张朦[2] 万有[2] 吴红海[3] 侯艳宁[3] 郑乐民[1] 尹玉新[1]
机构地区:[1]北京大学系统生物医学研究所,北京100191 [2]北京大学神经科学研究所,北京100191 [3]中国人民解放军白求恩国际和平医院药学部,石家庄050082
出 处:《分析化学》2015年第8期1118-1124,共7页Chinese Journal of Analytical Chemistry
基 金:国家自然科学基金项目(No.21305005)资助~~
摘 要:建立了高效液相色谱-串联质谱法(HPLC-MS/MS)测定鼠脑组织中6种神经甾体类激素(别孕烯醇酮、孕烯醇酮、孕酮、脱氧皮质酮、四氢脱氧皮质酮、去氢表雄酮)的方法。采用Agilent XDB-C18色谱柱,以5 mmol/L乙酸铵-甲醇为流动相,梯度洗脱分离,在电喷雾正离子模式下,以多反应监测(MRM)扫描模式采集数据,基质匹配内标法定量。孕酮和脱氧皮质酮在0.10~10 ng/m L范围内呈良好的线性关系,别孕烯醇酮、孕烯醇酮、四氢脱氧皮质酮、去氢表雄酮在0.50~10 ng/m L范围内呈良好的线性关系,相关系数均大于0.99;在0.4,4.0和8.0 ng/m L加标水平下的平均回收率为91.2%~115.5%;相对标准偏差(RSD,n=6)为0.87%~8.8%;检出限为0.04~0.20 ng/m L,定量限为0.10~0.50 ng/m L。An analytical method based on high performance liquid chromatography tandem mass spectrometry was developed for the simultaneous determination of 6 neurosteroids in rat brain tissues. The neurosteroids were separated on an Agilent XDB-Cls column (50 mm×4.6 mm, 1.8 μm), using methanol-5 mmol/L ammonium acetate as the mobile phase. A gradient elution program with a cycle time of 10 min was used. The neurosteroids were detected by electrospray ionization mass spectrometry in positive mode with multiple reaction monitoring (MRM) scan mode. For ll-deoxycorticosterone (DOC) and progesterone ( PROG), the linear range of calibration curve was from 0. 10 ng/mL to 10 ng/mL, and for allopregnanolone (AP), pregnenolone ( PREG), dehydroepiandrosterone (DHEA) and allotetrahydrodoe (THDOC), the linear range calibration curve was from 0.50 ng/mL to 10 ng/mL, both with the correlation coefficient more than 0.99. The mean recoveries at spiked levels of 0.4, 4.0 and 8.0 ng/mL were in the range of 91.2%-115.5% , and the RSDs were 0.9% -8.8% (n=6). The limits of detection and quantification were 0.04-0.20 ng/mL and 0.10-0.50 ng/mL, respectively.
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