基于衍生化HPLC法测定不同煎煮时间大叶黄精中18种游离氨基酸含量  

Pre-column Derivatization HPLC Determination of 18 Free Amino Acids of Polygonatum kingianum var.grandifolium with Different Decoction Times

作　　者：马宽 唐华丽[1] 邱顺丽 黄凯 李祥 马庆东 易东阳[2] Ma Kuan;Tang Huali;Qiu Shunli;Huang Kai;Li Xiang;Ma Qingdong;Yi Dongyang(Chongqing Three Gorges University,Chongqing 404100,China;Chongqing Three Gorges Medical College,Chongqing 404100,China)

机构地区：[1]重庆三峡学院,重庆404100 [2]重庆三峡医药高等专科学校,重庆404100

出　　处：《广东化工》2025年第1期161-163,132,共4页Guangdong Chemical Industry

基　　金：重庆市自然科学基金面上项目(cstb2022nscq-MSX1661);重庆三峡学院高层次人员科研启动项目(14RC06);重庆三峡医药高等专科学校科研苗圃项目(XJ2022003202);三峡库区地道药材开发利用重庆市重点实验室(Sys20210001)。

摘　　要：目的:建立了柱前衍生-高效液相色谱法测定大叶黄精中18种游离氨基酸含量,并评价其鲜品在不同煎煮时间下游离氨基酸含量的变化。方法:以异硫氰酸苯酯为衍生剂,采用XBridge■C18色谱柱(250 mm×4.6 mm,5μm),流动相A为1.64 g/L无水乙酸钠缓冲液,流动相B为80%乙腈水溶液,进行梯度洗脱,检测波长254 nm,流速1.0 mL/min,柱温40℃,测定不同煎煮时间大叶黄精中游离氨基酸含量。结果:经方法学考察,精密度、稳定性和重复性良好,各氨基酸在质量浓度为0.12μg/mL~115.2μg/mL范围内与峰面积呈良好的线性关系,r均≥0.9990,平均回收率在80.03%~108.85%之间;鲜品经煎煮8小时后总游离氨酸含量下降50.76%,其中亮氨酸、甘氨酸在煎煮0.5 h后含量下降十分显著,提示加热时间对大叶黄精鲜品的游离氨基酸总含量有显著影响。结论:本方法具有可行性,能较好地评价大叶黄精加工质量。Objective:To establish a pre-column derivatization-high performance liquid chromatographic method for the determination of 18 free amino acid content of Polygonatum kingianum var.grandifolium,and to evaluate the changes in amino acid content of its fresh products at different decoction times.Methods:Phenyl isothiocyanate was the derivatizing agent,the free amino acid content of Polygonatum kingianum var.grandifolium with different decoction times was determined on an XBridge®C18 column(250 mm×4.6 mm,5μm),the mobile phase A is 1.64 g/L anhydrous sodium,mobile phasea B is the cetate-80%acetonitrile aqueous solution in a gradient elution mode,with a detection wavelength of 254 nm,a flow rate of 1.0 mL/min,and a column temperature of 40℃.Results:After methodological investigation,the precision,stability and reproducibility were good,and each amino acid showed a good linear relationship with the peak area in the range of 0.12μg/mL~115.2μg/mL,with r≥0.9990,and the average recoveries ranged from 80.03%to 108.85%.The results showed that the total free amino acid content of the fresh title herb samples by decocting for 8 hours was decreased at 50.76%.Among of them,that the content of leucine and glycine decreased extremely after decocting for 0.5 h,suggesting that the heating time has a significant effect on the total free amino acid content of Polygonatum kingianum var.grandifolium during processing.Conclusion:This method is feasible and can evaluate the processing quality of Polygonatum kingianum var.grandifolium.

关 键 词：大叶黄精 游离氨基酸 柱前衍生化 高效液相色谱法 

分 类 号：O629.7[理学—有机化学] R283[理学—化学] R284[医药卫生—中药学] R917[医药卫生—中医学]