Thin layer chromatography quantitative analysis on gallic acid in Galla Chinensis based on image analysis technology
10.3760/cma.j.cn115398-20230417-00216
- VernacularTitle:基于图像分析的五倍子没食子酸薄层色谱定量研究
- Author:
Hongliang ZHOU
1
;
Yawei HUANG
;
Mingxin TIAN
;
Dengshan ZHANG
;
Yue HAN
Author Information
1. 南京中医药大学附属医院药学部，南京 210029
- Keywords:
Galla Chinensis;
Gallic acid;
Image J;
Image analysis;
Assay (TCD)
- From:
International Journal of Traditional Chinese Medicine
2024;46(3):363-368
- CountryChina
- Language:Chinese
-
Abstract:
Objectives:To discuss the feasibility of Image J in quantitative analysis on thin layer chromatography (TLC) using gallic acid in Galla Chinensis as research object.Methods:Silica gel GF 254 thin-layer plate was used with chloroform-ethyl formate-formic acid (5:5:1) as the developing solvent and the images were taken under ultraviolet light (254 nm). Polyamide film was used with 75% ethanol-glacial acetic acid (50:1) as the developing solvent and 1% ferric chloride ethanol solution as the chromogenic reagent, and the images were taken under sunlight. Images obtained from the above conditions were imported into Image J to analyze and calculate the content of gallic acid in Galla Chinensis by using external standard two-point method. High-performance liquid chromatography (HPLC) was used with a mobile phase of methanol-0.1% phosphoric acid solution (15:85) at a wavelength of 273 nm to determine the gallic acid content in Galla Chinensis. Results:The quantitation limit of gallic acid on silica gel GF 254 thin-layer plate was 0.401 6 μg, the linear range was 2.855 - 9.515 μg ( r2 = 0.996 0), and the average recovery was 105.12% ( RSD=3.48%); the quantitation limit of gallic acid on polyamide film was 0.363 4 μg, the linear range was 1.427 - 4.758 μg ( r2 = 0.991 5), and the average recovery was 103.75% ( RSD=4.60%). The HPLC method had a quantitative limit of 4.42 ng, a linear range of 0.122-0.977 μg ( r2 = 0.999 9), and a recovery rate of 98.30% ( RSD = 1.40%). The accuracy, repeatability and stability of RSD were all <5%. The gallic acid content measured using Image J showed a maximum relative error of 9.30% and a minimum of 1.62% compared to the HPLC results. Conclusions:Image J is feasible for quantitative analysis of TLC and can be used as a complementary method for quality control of Chinese materia medica.
