Studies On Material Foundation And Pharmacokinetics Of Ganning Granula

Ganning Granules'prescription is from the homemade Ganning oral liquid in a infectious diseases hospital. It is made from the Rhubarb and Chuanxiong. Ganning oral liquid is hospital self-make preparation, the larger dose, poor stability of quality, and no systemic quality standards. These is not suitable to large-scale production. To ensure efficacy, to reduce side effects and dosage, and to facilitate the production, storage, transportation and application, we developed the Ganning Granules on the basis of the prescription of the Ganning oral liquid and clinical application. The author believe, the Ganning Granules will provide a bright road for the rehabilitation of the majority of hepatitis patients when it is successfully developed.It's action are such as: promoting blood circulation to remove blood stasis, cooling the blood and relieving toxin, choleretic effect and reducing jaundice. It applies to acute hepatitis B and chronic liver diseases and hepatic cirrhosis, especially has obviously curative effect on hyper-bilirubinemia. It's aside action is little. According to the clinical application practice of the infectious disease hospital in many years, the prescription is proved to have better and very stable therapeutic effect on the acute and chronic liver disease and cirrhosis appetite. Combined anthraquinone with purgation from Rhubarb ferulic acid with anti-platelet aggregation, anti-inflammatory, antibacterial, antivirus effects from Chuanxiong is the main source treating acute hepatitis. Therefore, this paper studied the pharmacy and stability of the Ganning Granules with five combined anthraquinone and ferulic acid as the target. By the orthogonal test, we determine it's prescription and preparation technology, and establish the methods of studying stability. It offers the foundation for quality control in the preparation, storage, transport of the production. On the side, this paper also researched the pharmacokinetics of rhubarb anthraquinone derivatives in the Ganning Granules, and established the method to examine simultaneously the pharmacokinetics of the five anthraquinone derivatives. This method can directly reflect the concentration of the components in the blood samples, and gain variety of rhubarb anthraquinone derivatives'pharmacokinetic parameters. It provided the more adequate, strong scientific basis for clinical pharmacokinetics, pharmacology, toxicology and clinical application in reason. Ⅰ. Study on active part extraction and preparing of Ganning GanulaObjective: To study the technique of extracting active parts in each drug and preparing Ganning Granules. Method: Identifying the mainly direction of technology by reading the literatures, then orthogonal design test were done to find out the extraction process of each drug's effective ingredients, and select appropriate excipients to develop preparation. Result: Radix et Rhizoma Rhei was extracted with water and Rhizoma Chuanxiong was extracted with 80% alcohol to gain active ingredients. After the effective ingredients'extraction is concentrated and dried, it is made into Ganning Granules in the appropriate technology. Meanwhile we discovered instability of cis-isomers transformation of FA from Chuanxiong during the process of preparation for the first time. Conclusion: The preparation has a simple process, low cost, obvious efficacy, stable quality and is convenient to use. It is easily accepted by patients, and is suitable for industrilized production.Ⅱ. Study on stability of Ganning GranulaObjective: To study the law of the preparation changes with time's vary under influence of temperature and humidity and light ray. Method: To make use of different chromatographic conditions to quest the method on the concentration determination. According to the relevant guidelines of the Chinese Pharmacopoeia, the stress condition testing and accelerated tests and long-term trial was conducted to investigate the changes in characteristics, identification, moisture, granularity, content, and microbial limit. Results: We set up the TLC analysis methods of each herbal medicines and the HPLC of five anthraquinone combo(aloe-emodin, rhein, emodin, chrysophanol, physcion) and FA, and improved the HPLC of rhubarb and the TLC method of rhubarb and Chuanxiong in the Chinese Pharmacopoeia (2005 edition). In the stability test, except that the high humidity and the light ray slightly affect granularity, color, moisture and content of every ingredients in the stress condition testing, each experimental data and chromatograms in the accelerated tests and long-term trial are no significant change comparing to 0 month. Conclusion: Under present packing conditions, it's stability is better, and the prescription and the preparation technique are feasible essentially. The paper offers the scientific basis to production, packing, storage and transportation.Ⅲ. Study on pharmacokinetics of rhubarb anthraquinone in Ganning GranulaObjective: To establish a high performance liquid chromatography (HPLC) method on the Simultaneous determination of five rhubarb anthraquinones in rat plasma samples after Ganning Granules was administered to rats orally and to study on multicomponent pharmacokinetics in Ganning Granules by the use of plasma concentration measure. Method: After rat taking Ganning Granules suspension, the blood sampling was done according to a certain time point. The plasma was hydrolyzed by hydrochloric acid and extracted by ethyl acetate, and the plasma concentration of five rhubarb anthraquinone were mensurated by RP-HPLC. The conditions of chromatography was C18 column(4.6mm×150mm,μm), the mobile phase consisted of methanol-water-acetic acid (73:26.1:0.9), the flow rate of 1.0 ml/min, room temperature, the detection wave-length of 430nm. The pharmacokinetic parameters was calculated by the 3p97. Results: Aloe-emodin plasma concentration had good linear relationship within the concentration range of 0.0295μg/ml~3.78μg/ml, the correlation coefficient r=0.9997(n=7), the detection limit: 0.01μg/ml; Rhein plasma concentration had good linear relationship within the concentration range of 0.1188μg/ml~15.2μg/ml, the correlation coefficient r=0.9997(n=7), the detection limit: 0.05μg/ml; Emodin plasma concentration had good linear relationship in the concentration range of 0.0762μg/ml~3.25μg/ml, the correlation coefficient r=0.9994(n=5), the detection limit: 0.03μg/ml; Chrysophanol plasma concentration had good linear relationship in the concentration range of 0.1055μg/ml~13.5μg/ml, the correlation coefficient r=0.9994 (n=6), the detection limit: 0.05μg/ml; Physcion plasma concentration had good linear relationship in the concentration range of 0.75μg/ml~6.0μg/ml, the correlation coefficient r=0.9954(n=5), the limit of detection: 0.300μg/ml. From the pharmaco- kinetic parameters, the four tested indexes in the experiment belong all to the second compartment, and the speed of imbibition into the blood preliminarily, absorption and distribution is faster, but ejectable speed is slow relatively. In addition, a bimodal phenomenon of emodin was found, and the other three components were single peak. It provides a basis as an indication of the interaction between components in the compound preparation. Conclusion: This method is simple, sensitive, stable, accurate, and applies to determine plasma concentration of five rhubarb anthraquinone after Ganning Granules is administered to rats orally. On the side, the results lay the foundation of further clinical pharmacokinetics and pharmacokinetic-pharmaco- dynamic relevance.