LC-MS定量研究白桦脂酸的体外细胞摄取与转运  

LC-MS quantitative study on cellular uptake and transport of betulinic acid in vitro

作　　者：陈旺 王美慧 帅丽霞 詹雁[1] 谭镭[1] 阮佳[1] 徐超群[1] CHEN Wang;WANG Meihui;SHUAI Lixia;ZHAN Yan;TAN Lei;RUAN Jia;XU Chaoqun(Sichuan Provincial Academy of Chinese Medicine Science,Chengdu 610041,China;College of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China)

机构地区：[1]四川省中医药科学院,四川成都610041 [2]成都中医药大学药学院,四川成都611137

出　　处：《中国测试》2022年第3期59-65,共7页China Measurement & Test

基　　金：四川省重大科技专项(2017SZDZX005);国家重大新药创制项目(2018ZX09731013);四川省省级科研院所基本科研业务费项目(A-2020N-17)。

摘　　要：建立一种快速专一测定白桦脂酸(BA)含量的LC-MS方法,采用选择离子检测方式,测定Caco-2细胞中BA的含量,用于研究BA在Caco-2细胞中的摄取与转运情况。采用色谱柱Agilent SB-C18(2.1 mm×50 mm,1.8μm),流动相0.1%甲酸-甲醇(22∶78),流量0.3 mL/min。采用电喷雾离子源(ESI),负离子模式下定量分析BA和齐墩果酸(内标)碎片离子质荷比(m/z)455.4。BA在8.4~840 ng/mL范围内线性关系良好(r>0.996),定量限为8.4 ng/mL;日内、日间精密度RSD均小于10%,基质效应为107.9%~114.5%,提取回收率为97.2%~103.0%;样品室温、长期和冻融稳定性良好,RSD小于10%。该法快速、灵敏、简单、专属性强,可用于定量分析BA在Caco-2细胞的摄取与转运。To establish a rapid and specific LC-MS method to determine the concentration of betulinic acid in the Caco-2 cells and study the cellular uptake and transport of betulinic acid in vitro. The chromatographic analysis of Betulinic acid was performed on an Agilent SB-C18(2.1 mm×50 mm, 1.8 μm) with mobile phase0.1% formic acid water-methanol(22∶78) at a flow rate of 0.3 mL/min. The detection was performed by electrospray ion source(ESI) using the negative mode of selective ion monitoring(SIM) while the mass-tocharge ration(m/z) of betulinic acid and internal standard oleanolic acid were 455.4. The linear range of betulinic acid was 8.4-840 ng/mL with good linearity(r>0.996). The LLOD was 8.4 ng/mL. The intra-day and inter-day precisions RSD were below 10%, the matrix effect were 107.9% to 114.5%, and the recovery rate were 97.2% to 103.0%. The samples have good stability at room temperature, long-term and freeze-thaw, and its RSD was lower than 10%. The method was fast, sensitive, simple and specific, which could be used to study the cellular uptake and transport of betulinic acid.

关 键 词：白桦脂酸 LC-MS 细胞摄取 细胞转运 

分 类 号：R285.5[医药卫生—中药学]