Establishment of HPLC Fingerprint of Chinese Gall Leaven and Content Determination of 5 Components
- VernacularTitle:百药煎HPLC指纹图谱的建立及其中5种成分的含量测定
- Author:
Jingjing CHEN
1
;
Zhenling ZHANG
1
;
Miaomiao CAO
1
;
Ying ZHANG
1
;
Jianing YANG
1
Author Information
1. College of Pharmacy，Henan University of TCM，Zhengzhou 450008，China
- Publication Type:Journal Article
- Keywords:
Chinese gall leaven;
Fingerprint;
HPLC
- From:
China Pharmacy
2020;31(2):173-178
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE：To establis h HPLC fingerprint of Chinese gall leaven ，simultaneous determination of 5 components and optimization of distillers ’grains. METHODS ：HPLC method was adopted. The determination was carried out on Waters Symmetry Shield TM RP18 column with mobile phase consisted of acetonitrile- 0.1％ trifluoroacetic acid aqueous （gradient elution ）at the flow rate of 0.8 mL/min. The column temperature was 30 ℃，and detection wavelength was set at 280 nm. The sample size was 5 µL. Using gallic acid as reference ，HPLC fingerprints of 14 batches of sample were drawn. The similarity evaluation was performed by using Similarity Evaluation System of Chromatogram Fingerprint of TCM （2012 edition）to determine common peaks；SPSS 20.0 software was used for cluster analysis. RESULTS ：There were 11 common peaks in 14 batches of samples ，and the similarity range of 14 batches of samples was 0.424-0.998，A1-A5 was less than 0.850；5 components were identified in B 1-B9 sample，i.e. gallic acid ，（－）-epigallocatechin，methyl gallate ，2，4，6-tri-O-galloyl-α-D-glucose and 2，4，6-tri-O-galloyl- β-D-glucose. The results of cluster analysis showed that 14 batches of samples could be grouped into 3 categories，i.e. A1-A5 into one category，B1 and B 6 into one category ，B2-B5 and B 7-B9 into one category. The linear ranges of above 5 components were 29.96-599.2 μg/mL（r＝0.999 6），0.832-416 μg/mL（r＝0.999 6），0.102-51 μg/mL（r＝0.999 8），0.286 4-143.2 μg/mL（r＝0.999 8）， 0.286 4- 143.2 μg/mL（r＝0.999 8），respectively. The limits of quantitation were 0.060，0.104，0.017，0.029，0.057 μg/mL；the limits of detection were 0.018，0.031，0.005，0.009，0.017 μg/mL，respectively. RSDs of precision ，stability，reproducibility and durability tests were all lower than 3％. The recoveries were 97.16％-101.88％（RSD＝1.60％，n＝6），96.98％-99.24％（RSD＝ 0.85％，n＝6），97.7％-101.64％（RSD＝1.54％，n＝6），97.77％-103.08％（RSD＝1.82％，n＝6），98.16％-101.88％（RSD＝1.24％， n＝6），respectively. CONCLUSIONS：The established HPLC fingerprint and cluster analysis can be used to evaluate the quality of Chinese gall leaven. The established method is simple to Δ 基金项目：国家重点研发计划项目（No.2018YFC1707200）；公 operate and can be used to determine the contents of 5 益性行业科研专项项目（No.201507004-03）；河南中医药大学研究生 components simultaneously ；rice husk distillers ’grains were 科研创新基金项目（No.YJS2018B14） ＊硕士研究生 。研究方向 ：中药饮片及新药研究 。E-mail： not suitable for fermented Chinese gall leaven . 401327039@qq.com
