摘要
目的:建立柱前衍生化反相高效液相色谱法(RP-HPLC)同时测定不同产区锁阳中17种游离氨基酸含量的方法并进行多元统计分析。方法:采用柱前衍生化RP-HPLC检测,以异硫氰酸苯酯(PITC)为衍生化试剂,采用Wonda Sil C18-WR色谱柱(4.6 mm×150 mm,5μm),流动相A为0.05 mol·L^-1乙酸钠缓冲液(pH 6.5),流动相B为乙腈-甲醇-水(3∶1∶1),梯度洗脱,流速0.8 mL·min^-1,检测波长254 nm,柱温35℃。建立主成分分析(PCA)及系统聚类分析(HCA)模型进行多元统计分析并进行质量评价。结果:从锁阳中共分离鉴定了17种氨基酸,含7种人体必需氨基酸。17种氨基酸在各自质量浓度范围内呈良好线性关系,r在0.9990~0.9999;平均回收率在98.03%~103.89%,RSD均<3.5%。PCA和HCA结果基本一致,均可将12个市级产区明显区分为6个区域;PCA可根据不同市级或省级产区对锁阳进行聚类,而HCA可根据省级产区对锁阳进行归类,说明不同市级、省级产区锁阳的氨基酸含量具有产区差异性,且含量分布表现出明显的地域聚集性特点;PCA表明甘肃、内蒙古所产锁阳氨基酸含量较高,质量较佳。结论:所建方法可用于不同产区锁阳中17种游离氨基酸的含量测定,为其综合质量评价提供参考。
Objective:To establish a pre-column derivatization reverse-phase high-performance liquid chromatography(RP-HPLC)method for the simultaneous determination of 17 amino acids in Cynomorii Herba from different producing areas and conduct a multivariate statistical analysis.Method:RP-HPLC with pre-column derivatization was employed,with phenyl isothiocyanate(PITC)as derivatization reagent.Separation was performed on a WondaSil C18-WR column(4.6 mm×150 mm,5μm),with 0.05 mol·L^-1 sodium acetate solution(pH 6.5)as mobile phase A,and acetonitrile-methanol-water(3∶1∶1)as mobile phase B for gradient elution at a flow rate of 0.8 mL·min^-1.The detective wave length was set at 254 nm,and the column temperature was maintained at 35℃.Principal component analysis(PCA)and systematic cluster analysis(HCA)models were established for multivariate statistical analysis and quality evaluation.Result:17 Kinds of amino acid were detected in Cynomorii Herba,7 of which were essential amino acids.The 17 amino acids showed good linearity in respective concentration range,r=0.9990-0.9999.The average recoveries were between 98.03%-103.89%with RSD<3.5%.The results of PCA and HCA were basically the same,and both methods can be used to clearly distinguish Cynomorii Herba from 12 municipal producing areas into 6 regions.PCA can be used to classify Cynomorii Herba according to different municipal or provincial production areas,and HCA can be used to classify it according to provincial production areas.It showed that the amino acid contents in Cynomorii Herba from different municipal and provincial producing areas had differences,and the content distribution showed obvious geographical clustering characteristics.PCA showed that Cynomorii Herba from Gansu province and Inner Mongolia had higher amino acid contents and better quality as compared with other producing areas.Conclusion:The established method can be used for content determination of 17 amino acids in Cynomorii Herba from different producing areas,and provide a reference for its comprehensive quality evaluation.
作者
顾志荣
马转霞
马天翔
萨日娜
刘洁丽
吴国泰
祁梅
葛斌
GU Zhi-rong;MA Zhuan-xia;MA Tian-xiang;SA Ri-na;LIU Jie-li;WU Guo-tai;QI Mei;GE Bin(Gansu Provincial Hospital,Lanzhou 730000,China;College of Pharmacy,Gansu University of Chinese Medicine,Lanzhou 730000,China;Gansu Province Key Laboratory of Pharmacology and Toxicology of Traditional Chinese Medicine,Lanzhou 730000,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2020年第10期148-155,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
甘肃省中药药理与毒理学重点实验室开放基金项目(ZDSYS-KJ-2018-010)
甘肃省中药质量与标准研究重点实验室基金项目(ZYZL18-004)
甘肃省人民医院研发攻关项目(18GSSY2-3)。
作者简介
第一作者:顾志荣,硕士,主管中药师,从事中药分析与质量控制研究,E-mail:guzr8817@126.com;通信作者:祁梅,副主任药师,从事中药分析与质量控制研究,Tel:0931-8281933,E-mail:gssyqm@163.com;通信作者:葛斌,主任药师,从事中药分析与质量控制研究,Tel:0931-8281345,E-mail:gjy0630@163.com。
