Study On Quality Standard Of GanKang Capsule

Object:To establish a series of TLC methods to identify traditional Chinese medicinal herb in GanKang Capsule in order to provide one guideline for product quality control. To establish a series of high performance liquid chromatography methods for determination of the key chemical components in GanKang Capsule, for qulity control of the product.Methods:1、Studies on the methods to identify the GanKang Capsule by TLC:(1) Choose items for identification.(2) Choose the control articles for identification.(3) Optimization of the extraction method:different extraction methods, extraction solvents of different solvent composition and extration time were tested, for optimization of the effective extraction of test components from GanKang Capsule, further more avoidance interference.(4) Optimization of the thin layer chromatographic conditions:the thin layer chromatographic conditions such as static phase, composition and proportion of developing agent were tested for optimization of the chromatographic conditions.(5) Select appropriate colouration or examine conditions in order to make out the speckle clearly.(6) The test of selectivity:negative control samples were analyzed to evaluate the selectivity of the developed method.2、Studies on the methods to determination of Yi Gankang Capsule by HPLC:(1) Choose items for determination.(2) Select the effective extraction of test components from GanKang Capsule.(2) Choose appropriate chromatographic column, and adjust the formulation, proportion flow rate in order to separate the peaks of test components well.(3) Validated the methodology.(4) The established methods were applied for determination of Astragaloside IVglucoside, berberine hydrochloride, psoralen, emodin, chrysophanol and icariin in GanKang Capsule of different production batches. Results:1、Studies on the methods to identify the GanKang Capsule by TLC:(1) List the items of identification of Polyrhachis dives Smith, Astragalus monghbolicus(Bge.)Hsiao, Mahonia bealei(Fort)Carr., Ficus simplicissima Lour., Rumex japonicus Houtt., Epimedium brevicornum Maxim. into the quality standard of GanKang Capsule. Choose reference substance such as black ants control medicinal herb, Astragaloside IVglucoside, berberine hydrochloride, psoralen, emodin, chrysophanol and icariin as the control articles for identification. We didn’t list the items of identification of Ziziphus jujuba into the quality standard of GanKang Capsule, because there was interference in negative control in thin layer chromatogram during attempt.(2)The established method was applied for identification of Polyrhachis dives Smith, Astragalus monghbolicus(Bge.) Hsiao, Mahonia bealei(Fort) Carr., Ficus simplicissima Lour., Rumex japonicus Houtt., and Epimedium brevicornum Maxim. in GanKang Capsule. The result showed that the spots of the test solution were in the same position as those of the reference solution, without the interference of other chemical components originated from the coexisting herbs.2、Studies on the methods to determination of Yi Gankang Capsule by HPLC:(1) List the items of determination of Astragaloside IV, berberine hydrochloride, psoralen, emodin, chrysophanol and icariin into the quality standard of GanKang Capsule, which is grounded upon the prescription and quality standard principles of TCM.(2) Optimization of the extraction method. For determination Astragaloside Ⅳ, psoralen and icariin, GanKang Capsule was extracted with methanol by a sonifier for20min. For determination berberine hydrochloride, the preparation was extracted with acetonitrile-water(3:7) for30min and then ultrasound treatment for45min. For determination emodin and chrysophanol, the preparation was extracted with methanol by vibration wave for10min.(3)The chromatographic condition:The HPLC system was performed on a C18analytical column. The mobile phases for determination of Astragaloside Ⅳ, berberine hydrochloride, psoralen, emodin, chrysophanol and icariin were acetonitrile-water(30:70), acetonitrile-0.05mol/L potassium dihydrogen phosphate(30:70), methanol-water (45:55), methanol-0.05%phosphoric acid(85:15) and acetonitrile-water(30:70) separately. The wavelengthes were set at203nm,265nm,246nm,254nm and270nm separately.(4) Validated the methodology:The calibration curves of Astragaloside IV, berberine hydrochloride, psoralen, emodin, chrysophanol and icariin showed good linearity at the ranges of10.2-204μg/mL,22.5-225μg/mL,8.4-84μ/mL,25.6-256μg/mL,24.8-248μg/mL,7.05-141μg/mL. The reproducibilities of thses methods were good with RSD of1.61,1.54,1.76,2.88,1.06and1.24%, and the average recoveries were100.34,101.53,98.60,98.57,101.84,96.94%respectively. The contents of the six constituents should be more than7.6496μg,85.264fig,11.632μg,31.192μg,238.608μg,0.28mg respectively. The content of psoralen in Ficus simplicissima Lour. should be more than0.332%. The content of emodin and chrysophanol in Rumex japonicus Houtt. should be more than0.185%and0.92%.Conclusion:The developed TLC methods were proved to be accurate, sensitive and convenient, and can be used for the identification of GanKang Capsule, which provided some guideline for qulity control. The developed HPLC methods were accurate and selective and can be applied for determination of the key chemical components in GanKang Capsule for efficient quality control.