微透析联合LC-MS/MS法研究马钱子总碱囊泡凝胶透皮吸收

目的: 建立皮肤微透析采样技术联合LC-MS/MS同时测定马钱子碱和士的宁的分析方法,并研究大鼠经皮给予马钱子囊泡凝胶后的透皮吸收。 方法: 应用皮肤微透析技术对大鼠给药部位皮下进行采样;微透析样品以他克林为内标,不经预处理直接进样;色谱采用XDB-C₁₈柱,甲醇-乙腈-水(含0.05%甲酸和10 nmol·L^-1甲酸铵)梯度洗脱;质谱采用正离子扫描多反应监测(MRM)方式。,用于定量的离子对分别为m/z 335.2→m/z 184.2(士的宁),m/z 395.2→m/z 324.2(马钱子碱),m/z 199.1→m/ z 171.1(他克林)。 结果: 测定微透析样品中马钱子碱和士的宁2种成分的线性范围分别为0.195~50 ng·mL^-1和0.156~40 ng·mL^-1,日内和日间精密度(RSD)均小于15%,准确度、稳定性符合要求。 结论: 该方法操作简便、灵敏度高、专属性强,适用于皮肤微透析样品中马钱子碱和士的宁的测定。

Objective: To develop and validate a rapid,accurate,precise and highly sensitive LC-MS/MS method coupled with microdialysis sampling for quantitative analysis of brucine and strychnine after cutaneous application of Semen Strychni niosome gel in rat subjects. Method: Subcutaneous sampling was carried on microdialysis at a flow rate of 2 μL·min^-1.Samples were injected directly into LC-MS/MS with tacrine as internal standard(IS).Quantificative analysis was operated on XDB-C₁₈ column with gradient elution of acetonitrile-methanol-water(0.05% acetic acid and 10 nmol·L^-1 ammonium formate) and responses at the transitions m/z 335.2→m/z 184.2(strychnine),m/z 395.2→m/z 324.2(brucine) and m/z 199.1→m/z 171.1(tacrine),respectively,in positive electrospray ionization mode. Result: Calibration curves were linear over the range of 0.195-50 ng·mL^-1 for brucine and 0.156-40 ng·mL^-1 for strychnine.The intra-and inter-day precisions(RSDs) were lower than 15%.The accuracy and stability of the method met the requirement. Conclusion: The method is rapid,sensitive and specific for the determination of brucine and strychnine in microdialysis samples.

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