目的 以五倍子中没食子酸为研究对象，探讨Image J软件用于薄层色谱定量的可行性。 方法 采用硅胶GF254薄层板，以三氯甲烷-甲酸乙酯-甲酸（5∶5∶1）为展开剂，紫外光灯（254 nm）下拍照；采用聚酰胺薄膜，以75%乙醇-冰醋酸（50∶1）为展开剂，1%三氯化铁乙醇溶液为显色剂，日光下拍照。将图像导入Image J软件进行分析，以外标两点法计算五倍子中没食子酸含量。采用HPLC法，以甲醇-0.1%磷酸溶液（15∶85）为流动相，在273 nm波长下测定五倍子中没食子酸含量。 结果 硅胶GF254薄层板没食子酸定量限为0.401 6 μg，线性范围2.855~9.515 μg（r 2=0.996 0），回收率为105.12%（RSD=3.48%）；聚酰胺薄膜没食子酸定量限为0.363 4 μg，线性范围1.427~4.758 μg（r 2=0.991 5），回收率为103.75%（RSD=4.60%）；HPLC法没食子酸的定量限为4.42 ng，线性范围0.122~0.977 μg（r 2=0.999 9），回收率为98.30%（RSD=1.40%），3种方法的精密度、重复性、稳定性RSD值均<5%。使用Image J测得的没食子酸含量与HPLC测定结果比较，最大相对误差9.30%，最小相对误差1.62%。 结论 Image J软件可用于薄层色谱定量分析，可作为中药质量控制方法的补充。 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 GF254 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 GF254 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.
Thin layer chromatography quantitative analysis on gallic acid in Galla Chinensis based on image analysis technology
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 GF254 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 GF254 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.
万方数据
