摘要
目的 建立紫背万年青水提和醇提物给药后非小细胞肺癌A549细胞中的乳酸含量的超高效液相色谱-质谱法测定方法,评价上述提取物的治疗活性。方法 超高效液相色谱-质谱分析使用ESI离子源,正离子扫描,MRM模式,色谱柱为Waters ACQUITY UPLC BEH C_(18)(100 mm×2.1 mm,1.7μm),流动相为含0.1%甲酸的水溶液和含0.1%甲酸的乙腈溶液;细胞实验使用非小细胞肺癌A549细胞,分别加入不同浓度(20~80μg·mL^(-1))的紫背万年青水提和醇提物,培养24 h后进UPLC-MS测定乳酸浓度。结果 乳酸的UPLC-MS检测方法线性良好,能够对2~1 000μg·mL^(-1)浓度的乳酸进行准确测定,最低定量限为1 ng·mL^(-1),回收率为92.79%~108.57%,RSD为4.48%~10.05%,实测样本结果显示,与对照组比较,醇提物对乳酸含量影响较水提物更大。结论 本研究阐明了不同溶剂制备的紫背万年青提取物对非小细胞肺癌A549细胞中的乳酸含量的影响,并建立了快速、灵敏、大通量的UPLC-MS测定方法。
OBJECTIVE To establish an ultra-high performance liquid chromatography-mass spectrometry(UPLC-MS) method for the determination of lactic acid in non-small cell lung cancer A549 cells after administration of aqueous extract and alcohol extract ofdieffenbachia purpura,and to evaluate the therapeutic activity of the extracts.METHODS Waters ACQUITY UPLC BEH C_(18)(100 mm×2.1mm,1.7μm) column was used for ultra-performance liquid chromatography-mass spectrometry.The electrospray ionization was used in positive ion mode for MRM mode.The mobile phase consisted of 0.1% formic acid aqueous solution and 0.1%formic acid acetonitrile solution.Non-small cell lung cancer A549 cells were cultured with aqueous extract and alcohol extract of dieffenbachia purpura of different concentrations(20-80 μg·mL~(-1)).After 24 h culture,lactic acid concentration was determined by UPLC-MS.RESULTS The UPLC-MS method for lactic acid detection had good linearity,and could accurately determine lactic acid at the concentration of 2-1 000 μg·mL~(-1).The lowest limit of quantification was 1 ng·mL~(-1),and the recovery rates were 92.79%-108.57%,RSD was 4.48%-10.05%.The results of the measured samples showed that the effect of alcohol extract on lactic acid content was greater than that of water extract.CONCLUSION This study elucidates the effects of different solvents on the content of lactic acid in non-small cell lung cancer A549 cells,and establishes a rapid,sensitive and high-throughput UPLC-MS method for determination.
作者
洪笃云
程良凯
HONG Duyun;CHENG Liangkai(Guangdong Second Provincial Hospital of T.C.M.(Guangdong Province Engineering Technology Research Institute of T.C.M.),Guangzhou,Guangdong 510095,China)
出处
《今日药学》
CAS
2023年第12期911-913,927,共4页
Pharmacy Today
基金
广东省中医药局科研项目(20202016)。
