液质联用分析芦丁、野黄芩苷和半枝莲碱B在小鼠灌胃给药后体内的组织分布

目的：建立UPLC-MS同时测定小鼠组织中芦丁、野黄芩苷和半枝莲碱B的分析方法，并对该3种成分在小鼠体内的组织分布进行研究。方法：以2-氰基-3，12-二氧代齐墩果烷-1，9（11）-二烯-28-酸甲酯（CDDO-Me）为内标，通过甲醇沉淀组织样品蛋白，使用LC-MS技术进行定量分析，采用ACQUITY UPLC BEH-C₁₈色谱柱（100 mm×2.1 mm，1.7 μm），以0.1%甲酸水溶液-乙腈为流动相，梯度洗脱，正、负离子检测模式，检测离子质荷比（m/z）为芦丁609.2、野黄芩苷463.3、半枝莲碱B558.4和CDDO-Me 506.7。结果：在小鼠肝脏、心脏、脾脏、肺脏、肾脏等组织中，芦丁、野黄芩苷、半枝莲碱B分别在0.005 0~0.160 0、0.004 0~0.133 0、0.003 3~0.106 7 μg·mL^-1的线性范围内线性关系良好（r≥0.991 3）；方法的精密度和稳定性RSD均小于15%；回收率为90.0%~98.4%，RSD小于15%，符合化学药物非临床药代动力学研究技术指导原则。芦丁等3个成分在小鼠体内分布比较广泛，其中在肝脏和肾脏中分布较多，在肺脏中分布含量最低。结论：本方法经方法学验证，可对小鼠组织中的芦丁、野黄芩苷、半枝莲碱B含量进行测定；在用于小鼠体内化学成分的组织分布研究中，芦丁在肾脏中含量多、脾脏中最少，野黄芩苷在心脏中含量最多、肺脏中最少，半枝莲碱B在肝脏中含量最多、肾脏中最少。

Objective: To establish a UPLC-MS method for simultaneous determination of rutin, scutellarin and scutebarbatine B in mice tissues and apply the method to tissue distribution study of these three components. Methods: The tissue samples were precipitated with methanol and analyzed by LC-MS with 0.1% formic acidacetonitrile solution as the gradient elution mobile phase. The separation was performed on an ACQUTITY UPLC BEH-C₁₈ (100 mm×2.1 mm, 1.7 μm), and 2-cyano-3, 12-dioxo Oleanane-1, 9 (11)-28-diene acid methyl ester (CDDO-Me)was used as an internal standard. The mass spectrometer was operated in positive and negative electrospray ionization modes,and the transition m/z 609.2,m/z 463.3,m/z 558.4 and m/z 506.7 were used for rutin, scutellarin,scutebarbatine B and CDDO-Me,respectively. Results: Excellent linearity (r≥0.991 3) was observed in all three studied components within their linear ranges,with 0.005 0-0.160 0 μg·mL^-1 for rutin,0.004 0-0.133 0 μg·mL^-1 for scutellarin,and 0.003 3-0.106 7 μg·mL^-1 for scutebarbatine B. The RSD of precision and stability of the developed method was less than 15%. The recovery of the method was 90.0%-98.4% with RSD less than 15%, in line with the technical guidelines for non-clinical pharmacokinetic study of chemical drugs. Rutin, scutellarin and scutebarbatine B were widely distributed in mice tissues. The majority was detected in liver and kidney,and the lowest content was in lung. Conclusion: The method is proved by methodological validation that it can be used for content determination of rutin,scutellarin and scutebarbatine B in mice tissues. Rutin was detected mostly in kidney and the lowest was found in spleed,scutellarin was detected mostly in heart and the lowest was found in lung,and scutebarbatine B was detected mostly in liver and the lowest was found in kidney.