Quality Standard Revise And Finger Print Study Of Pipazhike Granule

Pipazhike Granule is an OTC drug used for cough-relieving, which is consisted of folium eriobotryae, sessile stemona root tuber, opium poppy peel, cynanchum stauntoni, cortex mori radicis, balloonflower root and menthol. Pipazhike granule had the activity of clearing heat, sputum-removing and cough-relieving, it is a natural herbal product used to treating common cold and cough, which is always used to cure cough and Bronchitis clinically. Pipazhike Granule is enrolled into ministry authorized standard (WS3-B-1145-92), but there are only contain the quantitation of Codeine Phosphate by Thin Slice Scan and the qualitation of alkaloidal in opium poppy peel, and all other herbs lack of identification method, so the level of quality standard is low.In order to warrant the effectiveness, safety and quality control of Pipazhike granule, we renew the identification and content determination methods of Pipazhike granule. In our study, we firstly built and revised the TLC identification method of folium eriobotryae, opium poppy peel, menthol, cortex mori radicis, sessile stemona root tuber, balloonflower root and cynanchum stauntoni, and then revised the HPLC quantification method of codeine phosphate. On the base of above study, we futher the finger print study of Pipazhike granule, and built the research method of HPLC/UV finger print of Pipazhike granule. Our study gave a solid base for the quality contol and safety use of Pipazhike granule.There are two parts in our present study:1. The quality standard revisement of Pipazhike granule In this study, we tried to revise the quality standard of Pipazhike granule. We identified folium eriobotryae, opium poppy peel, menthol, cortex mori radicis, sessile stemona root tuber, balloonflower root and cynanchum stauntoni in Pibazhike granule by TLC, the method we built could identify sessile stemona root tuber, cynanchum stauntoni and Codeine Phosphate, Morphine Hydrochloride and Papaverine Hydrochloride in opium poppy peel. These methods were of high specificity without negative control interferences, were of high convenience and reproductivity, and could be used for the quality control of Pibazhike Granule.We also built the HPLC method to determinate codeine phosphate in Pipazhike granule, we used shiseido C18 colume (250 mm×4.6 mm,5μm),the mobile phase was 0.1mol/L sodium dihydrogen phosphate-methol(85:15),the detect wave length was 238 nm,the colume temperature is 30.0℃. The linear range of codeine phosphate was 0.0307～3.0672μg,the average coefficient of recovery was 99.43%,the RSD was 0.89 (n=6). This method was convient, with high sensitivity, high reproductivity, which should be acted as index of quality control. 2. The finger print study of Pipazhike granuleIn view of Pipazhike Granule was a compound preparation, and many herbs lacked of identificaion methods and the assaying method of acitve compounds. In our study, we tried to build the HPLC/UV finger print of Pipazhike Granule, elavated the quality control of Pipazhike Granule. We used octadecylsilane chemically bonded silica as loading agent (length of column: 25cm; inner diameter: 4.6mm, particle diameter: 5μm), the mobile phase is methanol-0.03mol/L sodium acetate (set the pH=3.5 by glacial acetic acid, 35:65), measurement wave length: 238nm, column tempreture: 30℃, flow rate:1.0ml/min. theoretical plate number was not low than 10000 calculated used the peak of Codeine Phosphate. We took the solutions Pipazhike Granule 12g, dissolved in caustic soda solution (0.5mol/L), standing for 30 min, transferred into separatory funnel. The bottle was washed several times by caustic soda solution (0.5mol/L) and the washings were also transferred into separatory funnel. The solutions were extracted by chloroform for 4 times, 40 ml each time, merged all the extractions, evaporated to dryness, the residue were dissolved into mobile phase, transferred into volumetric flask and added mobile phase to 25ml, then agitated, filtrated and the filtrate was used as reference solutions. We dropped test solutions and reference solutions 10μl each, injected into HPLC, recorded the chromatogram, and determined 8 peaks as the owned peaks of finger print. We choosed 5 peaks as characteristic peaks, and used them to calculate mached data, semblance and the relative retention time and relative peak aera of samples, all of these were used as technical parameter to evaluate finger print. The finger print we built was of high characteristic, and we identified the HPLC/UV peak ascription of Pipazhike granule, our study gave a solid base for further study on the spectrum-efficacy study of Pipazhke granule.In conclusion, we firstly identified sessile stemona root tuber, cynanchum stauntoni and Codeine Phosphate, Morphine Hydrochloride and Papaverine Hydrochloride in opium poppy peel by TLC and determinate codeine phosphate by HPLC method. These methods were of high convenience and reproductivity, high specificity without negative control interferences, and could be used for the quality control of Pipazhike Granule. Secondly, we built the finger print of Pipazhike Granule by HPLC/UV, and got the Reference Characteristic Chromatogram. There were five characteristic peaks in Reference Characteristic Chromatogram, which might exhibit the characteristic of principal active compounds, might be useful in the quality control of Pipazhike Granule.