Pre-column Derivatization RP-HPLC for Simultaneous Determination of 17 Amino Acids in Cynomorii Herba from Different Producing Areas and Its Multivariate Statistical Analysis
10.13422/j.cnki.syfjx.20200711
- VernacularTitle:柱前衍生化RP-HPLC同时测定不同产区锁阳中17种游离氨基酸含量及其多元统计分析
- Author:
Zhi-rong GU
1
;
Zhuan-xia MA
2
;
Tian-xiang MA
2
;
Ri-na SA
1
;
Jie-li LIU
2
;
Guo-tai WU
3
;
Mei QI
1
;
Bin GE
1
Author Information
1. Gansu Provincial Hospital, Lanzhou 730000, China
2. College of Pharmacy, Gansu University of Chinese Medicine, Lanzhou 730000, China
3. Gansu Province Key Laboratory of Pharmacology and Toxicology of Traditional Chinese Medicine, Lanzhou 730000, China
- Publication Type:Research Article
- Keywords:
Cynomorii Herba;
amino acid;
PITC;
pre-column derivatization;
RP-HPLC;
Principal component analysis;
systematic cluster analysis;
quality evaluation
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2020;26(10):148-155
- CountryChina
- Language:Chinese
-
Abstract:
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, 5 μm), 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 = 0.999 0-0.999 9.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.
