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目的建立HPLC-TQ-MS/MS分析方法同时测定大鼠血浆中的小檗碱、大车前苷、柴胡皂苷a、延胡索乙素、芍药苷和苦杏仁苷,并用于研究大鼠口服加味败藤浸膏后的药动学。方法色谱柱为Agilent Poroshell 120EC-C18柱(3.0 mm×100 mm,2.7μm),流动相为0.05%甲酸-乙腈(含0.05%甲酸),按程序梯度洗脱。质谱检测方式为多反应离子监测(MRM),采用正、负离子模式切换及时间分段扫描方法。用于定量分析的离子反应分别为m/z 335.9→m/z 319.9(小檗碱,ESI+),m/z 639.1→m/z 160.9(大车前苷,ESI-),m/z 779.3→m/z 617.4(柴胡皂苷a,ESI-),m/z 356.0→m/z 192.0(延胡索乙素,ESI+),m/z 449.1→m/z 121.1(芍药苷,ESI-),m/z 456.1→m/z 323.1(苦杏仁苷,ESI-),m/z 323.9→m/z 126.9(格列齐特,内标,ESI+)和m/z 321.9→m/z 170.1(格列齐特,内标,ESI-)。大鼠单剂量口服加味败藤浸膏(3.1 g·kg-1)后眼底静脉丛取血,血浆经甲醇预处理后进入HPLC-TQ-MS/MS分析。用DAS统计软件计算加味败藤浸膏中活性成分的药动学参数。结果经方法学考证,血浆中小檗碱、大车前苷、柴胡皂苷a、延胡索乙素、芍药苷和苦杏仁苷的线性范围分别为0.59~292.50,0.68~168.75,6.05~1 512.50,0.68~337.50,6.70~1 675.00和5.60~1 400.00 ng·mL~(-1),定量限分别为0.59,0.68,6.05,0.68,6.70和5.60 ng·mL~(-1),精密度、回收率及稳定性均良好,符合生物样本的分析要求。结论本实验所建HPLC-TQ-MS/MS分析方法灵敏、简便、可靠,适用于加味败藤浸膏的药动学研究。
OBJECTIVE To establish a HPLC-TQ-MS / MS method for the simultaneous determination of berberine, sarcosine, saikosaponin a, tetrahydropalmatine, paeoniflorin and amygdalin in rat plasma and to study the effects of oral dosing Pharmacokinetics after the rotten vine extract. Methods The column was Agilent Poroshell 120EC-C18 column (3.0 mm × 100 mm, 2.7 μm). The mobile phase consisted of 0.05% formic acid-acetonitrile (containing 0.05% formic acid). MS detection method for the multi-reactive ion monitoring (MRM), using positive and negative ion mode switching and time segmentation scanning method. The ion reactions for quantitative analysis were m / z 335.9 → m / z 319.9 (berberine, ESI +), m / z 639.1 → m / z 160.9 m / z 617.4 (saikosaponin a, ESI-), m / z 356.0 → m / z 192.0 (tetrahydropalmatine, ESI +), m / z 449.1 → m / z 121.1 z 456.1 → m / z 323.1 (amygdalin, ESI-), m / z 323.9 → m / z 126.9 (Gliclazide, internal standard, ESI +) and m / z 321.9 → m / z 170.1 Special, internal standard, ESI-). After single oral dose of dexamethasone extract (3.1 g · kg -1), rats were sacrificed and blood was collected from the venous plexus of rats. The plasma was pretreated with methanol and then entered into HPLC-TQ-MS / MS. Calculate the pharmacokinetic parameters of the active ingredient in the flavored Chinese cabbage extract with DAS statistical software. Results The methodological study showed that the linear range of berberine, stilbene glycoside, saikosaponin a, tetrahydropalmatine, paeoniflorin and amygdalin were 0.59 ~ 292.50,0.68 ~ 168.75,6.05 ~ 1 512.50,0.68 ~ 337.50,6.70 ~ 1675.00 and 5.60 ~ 1 400.00 ng · mL ~ (-1), respectively. The limits of quantification were 0.59,0.68,6.05,0.68,6.70 and 5.60 ng · mL -1, respectively. The precision and recovery And stability are good, in line with the biological sample analysis requirements. Conclusion The HPLC-TQ-MS / MS analysis method established in this experiment is sensitive, simple and reliable and is suitable for the study of pharmacokinetics of Modified Cabernet Sauvignon.