ABSTRACT
Objective To establish a method for the simultaneous determination of Forsythoside A, Phillyrin, (R,S)-Epigoitrin, Chlorgenic Acid and Isochlorogenic Acid A by HPLC, and test 16 batches of samples from 14 manufacturers. Methods The test was performed on Kinetex EVO C18 column (150 mm × 4.6 mm, 5 μm) with the column temperature at 35 ℃ . The gradient elution was adopted with the mobile phase of acetonitrile and 0.3% phosphoric acid aqucous at a flow rate of 1.0 ml/min. The detection wavelength of (R,S)-Epigoitrin and Phillyrin were set as 236 nm, the detection wavelength of Forsythoside A, Cholorogenic Acid and Isochlorogenic Acid A were set as 327 nm. Results The good linear relationships were displayed within the linear range of 0.050 45-2.018 00 μg for Forsythoside A (r=0.999 9), 0.018 21-0.728 40 μg for Phillyrin (r=0.999 9), 0.010 16-0.406 40 μg for (R,S)-Epigoitrin (r=0.999 9), 0.006 60-0.263 90 μg for Cholorogenic Acid (r=0.999 9) and 0.0040 44~0.161 76 μg for Isochlorogenic Acid A ( r=0.999 5). The RSDs of reproducibility and stability tests were lower than 2%; recoveries were 97.01%, 98.28%, 99.35% and 96.21%, RSD were 3.19%, 1.19%, 0.81%, 2.88% and 2.96%. The content ranges of Forsythoside A, Phillyrin, (R,S)-Epigoitrin, Chlorgenic Acid and Isochlorogenic Acid A from 16 batches of samples from 14 manufacturers were 0.057 43-1.508 71 mg/g, 0.017 72-0.350 15 mg/g, 0.005 68-0.177 13 mg/g, 0.007 53-0.226 33 mg/g and 0.00308-0.11908 mg/g. Conclusions The established method is simple and accurate, and has a good repeatability. It can be used for the quality analysis of Forsythoside A, Phillyrin, (R,S)-Epigoitrin, Chlorgenic Acid and Isochlorogenic Acid A. The content of the tested chemical components from 16 batches of samples from 14 manufacturers have significant differences which indicate that a reinforcement of the quality control is needed.
ABSTRACT
Objective To establish HPLC method for determination of (R,S)-epigoitrin in Banlangen granules and discuss the content limitation. Methods The samples were separated on SHIMADZU VP-ODS (150 mm×4. 6 mm, 5 μm) and the mobile phase consisted of methol-0. 02% phosphoric acid solution (7:93) at a flow rate of 1. 0 mL. min-1 . The detection wavelength was set at 245 nm and the injection volume (in automatic sampler) was 10 μL. The content limitation was assessed according to the transfer rate and statistical data of the results. Results (R,S)-epigoitrin showed a good linear relationship at concentration of 0. 058 7 – 150. 349 5 μg·mL-1(r = 0. 999 7, n = 7). The average recovery rate was 98. 72% , 98. 40% and 98. 60% , respectively; RSD was 1. 84% , 0. 50% and 1. 82% , respectively. The content limitation of (R,S)-epigoitrin was unreasonable according to the transfer rate and the statistical data of the results. Conclusion The method is easy and simple to operate, accurate and stable in results, and highly specific, thus it is applicable for the quality control of Banlangen granules. The content limitation should be determined on the basis of pharmacokinetic and pharmacodynamic data.