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首页> 数字期刊群 >本期导读>LC-MS定量研究白桦脂酸的体外细胞摄取与转运

LC-MS定量研究白桦脂酸的体外细胞摄取与转运

77    2022-07-01

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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 Wang1,2, WANG Meihui1,2, SHUAI Lixia1,2, 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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