ABSTRACT
Objective: To establish the quality control method for Juhuang Oral Liquid (JOL) based on the qualitative and quantitative analysis methods. Methods: TLC was used to identify Lycii Fructus and Polygonati Rhizoma in JOL. Phenol-sulfuric acid method was used for the detemination of total polysaccharide, HPLC was used to simultaneously determine the contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, coffeic acid, galuteolin, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, and 4,5-dicaffeoylquinic acid. The Odyssil C18 (250 mm × 4.6 mm, 5 μm) column was used. The mobile phase was acetonitrile-0.05% phosphoric acid solution in gradient elution, at the flow rate of 1.0 mL/min, the detection wavelength was set at 325 nm. Results: The identified characteristics of Lycii Fructus and Polygonati Rhizoma by TLC were distinct and the spots were clear, without interference and easy to recognize. When phenol-sulfuric acid method was used to determinate the total polysaccharide content, the regression equation was A=0.057 8 C + 0.032 5 (r=0.999 1, n=8). The linear with its absorbance in the range of 26.15-196.13 mg/mL, the average recoveries was 99.82%. The average content of the total polysaccharide in JOL from six batches is 157.74-166.49 mg/mL. When RP-HPLC was used to determine the eight chemical components, the compounds showed a good linearity(r ≥ 0.999 8) in the determination range. The average recoveries were between 99.18%-101.06% with RSDs between 0.79%-1.91%. The average content from the six batches showed neochlorogenic acid 102.9-142.6 μg/mL, chlorogenic acid 488.6-563.8 μg/mL, cryptochlorogenic acid 58.9-71.6 μg/mL, coffeic acid 240.7-326.1 μg/mL, galuteolin 228.6-302.7 μg/mL, 3,4-dicaffeoylquinic acid 398.0-485.4 μg/mL, 3,5-dicaffeoylquinic acid 184.1-203.1 μg/mL, and 4,5-dicaffeoylquinic acid 476.2-561.8 μg/mL. Conclusion: TLC identification method is simple. The determination method of total polysaccharide and chemical components is simple, accurate, and with a good repeatability. It can be used for the quality control of JOL.
ABSTRACT
OBJECTIVE: To establish an HPLC method for simultaneous determination of the contents of Chlorogenic Acid and Coffeic Acid in the Fruit of Chaenomeles. METHODS: HPLC was applied to determine and compare the contents of 11 batches of Chaenomeles medicinal materials collected at different time, prepared with different process and stored for different length of time, using Coffeic Acid and Chlorogenic Acid standard substances. The chromatogram conditions were as follows: Kromasil C18( 250mm? 4. 6mm, 5? m) column; mobile phase of ( A) acetonitrile, ( B) 0. 05% phosphoric acid; gradient el-ution ( 0～ 10min, 0% A～ 15% A, 100% B～ 85% B; 10～ 40min ( 15% A～ 30% A , 85% B～ 70% B) ; flow rate at 0. 8 mL? min- 1; detection wavelength at 325nm. RESULTS: The content of Chlorogenic Acid in batches 1 to 9 were respectively 0. 020% , 0. 035% , 0. 043% , 0. 051% , 0. 260% , 0. 104% , 0. 081% , 0. 056% , 0. 034% ; that of Coffeic Acid were respectively 0. 005% , 0. 006% , 0. 008% , 0. 010% , 0. 051% , 0. 024% , 0. 020% , 0. 015% , 0. 007% . The contents of Chlorogenic Acid and Coffeic Acid in batch 8’ ( burn- prepared) were respectively 0. 045% and 0. 010% ; those of Chlorogenic Acid and Coffeic Acid in batch 9’ ( stored for 1a) were respectively 0. 023% and 0. 005% . CONCLUSIONS: The method is simple, reproducible and suitable for the quality control of Fruit of Chaenomeles. The best time for harvesting batch 5 is on July 12, and the best process for this batch is solarization. The storage time has certain influence on the content of Chlorogenic Acid.