作者：陈旺^1,2, 王美慧^1,2, 帅丽霞^1,2, 詹雁¹, 谭镭¹, 阮佳¹, 徐超群¹

作者单位：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

关键词：白桦脂酸;LC-MS;细胞摄取;细胞转运

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，1.8 μm），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。

LC-MS quantitative study on cellular uptake and transport of betulinic acid in vitro
CHEN Wang^1,2, WANG Meihui^1,2, SHUAI Lixia^1,2, ZHAN Yan¹, TAN Lei¹, RUAN Jia¹, XU Chaoqun¹
1. Sichuan Provincial Academy of Chinese Medicine Science, Chengdu 610041, China;
2. College of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China
Abstract: 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 phase 0.1% 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-to-charge 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.
Keywords: betulinic acid;LC-MS;cellular uptake;cellular transport
2022, 48(3):59-65  收稿日期: 2021-01-27;收到修改稿日期: 2021-03-16
基金项目: 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）
作者简介: 陈旺（1996-），女，四川大竹县人，硕士研究生，专业方向为中药新制剂、新剂型、新技术应用研究
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