Analysis Of Multi-index Components Of Chinese Patent Medicine And Its Application In Quality Control

Chinese patent medicine (CPM) is widely used in clinic, which was directed by the theory of traditional Chinese medicine (TCM). It is composed of many medical herbs and made according to a rated prescription and technology. Nowadays, the model of quality control of CPM usually imitated chemical drugs. It is overwhelmed for almost half a century except the alternation of the analysis method and the index component analyzed. The therapeutic effect of TCM, especially CPM, is not a simple merging of each herb. In fact, it is the synergistic effect of each herb so it is more and more deficient to apply the chemical drugs'model of quality control to evaluate CPM. According to TCM theory, each herb of the prescription can affect the action of CPM, the quantity and proportion of the herbs can also affect its effect. It is a good method to control the quality of CPM by the analysis of multi-index components. Moreover, the method not only reflects the level of the production technics of CPM and the quality of each herb, but also reflects the condtion of its compatibility. But due to the particularity of CPM, such as complex components, larger difference of dissolubility, lower content of some index components, so that it is difficult to select appropriate extraction method and establish specific method for the simultaneous determination of multi-index components.A system of multi-index components analysis has been established, which included choosing the pretreat methods, optimizing the analytical method, evaluating dissolution and studying pharmacokinetics. The contents of multi-index components of some CPM were detected by the system, such as such as Sanhuang tablets of different manufactories, Gonglao Quhuo tablets and Qingwei Huanglian tablets. At the same time, the dissolution degree and pharmacokinetics of Gonglao Quhuo tablets were investigated. From the study, it can be concluded: (1) The method established for determine the contents of multi-index components of CPM are efficient, sensitive and specific (2) The obvious differences was observed by determining the index components of different companies'Sanhuang tablets. Therefore, it is more rational to control the quality of CPM by supervising of multi-index components. (3) The dissolution degree of mult-index of Gonglao Quhuo tablets was studied, to provide basis for appraisal and control the quality of CPM. (4) The compatibility of complex prescription can affect some important pharmacokinetics parameters in vivo. A good correlation was achieved by detecting Gonglao Quhuo tablets, which was used to evaluate internal predictability of correlation in vitro and in vivo. PART 1 Study of the Extraction Method and Extractive Solvent of Multi-index Components in CPM(1) Study of the influence of the Extraction Rate of 5 Rheum anthraquinone in Sanhuang tablets by Different Extraction TechniquesObjective: The effect of surfactant extraction on the extraction rate of 5 kinds of anthraquinones in Sanhuang tablets.Methods: Extration techniques using methanol circum- fluence, ultrasonic extraction and ultrasonic extraction by different concentration of SDS extraction were compared for the extration rate of 5 kinds of anthraquinones. The extrations were analyzed by HPLC.Results: The anthraquinone was extracted by ultrasonics, using 0.5% SDS as solvate; it could greatly improve the extraction rate.Conclusion: The solution of SDS was more effective than that of the conventional methods.(2) Studies on the extraction method and extractive solvent of Qingwei Huanglian tabletsObjective: To develop the best extraction method and extractive solvent of Qingwei Huanglian tablets, which provided basis for appraisal and control the quality of CPM. Methods: A extraction tset system was established by studying the extraction condition in different extraction method and extractive solvent of Qingwei Huanglian tablets.Results: The quantity extraction of 4 index components of Qingwei Huanglian tablets were siginficantly improved by ultrasonic extracting method with methanol.Conclusion: The extraction system was suitable for extracting the multi-index components of Qingwei Huanglian tablets.PART 2 The Study of the Method for Determination the Contents of the Multi-index Components of CPM(1) Determination the Contents of 4 index Components of Sanhuang Tablets from Different CompaniesObjective: To establish an RP-HPLC method for simulta- neous determination the contents of 4 index components (rhein, emodin, baicalina and berberine hydrochloride) of sanhuang tablets, and to evaluate the quality of the tablets from different companies.Methods: A good separation was achieved on a Kromasil C18 (4.6 mm×250 mm, 5μm) column at room temperature. The mobile phase was consisted of methanol, acetontrile and 0.1% triethylamine (pH 3.0) with gradient elution at the flow rate of 1.0 mL·min^-1. The UV detection wavelength was set at 254 nm.Results: The good linearity of Rhein, Emodin, Baicalina and Berberine Hydrochloride (r≥0.999 8) was achieved, the average recoveries of the method were 99.8%, 100.2%, 100.3% and 98.1%, respectively.Conclusion: The method is rapid, accurate and reliable. It was suitable for quality control of sanhuang tablets. Quality differences were found in sanhuang tablets from different companies.(2) Simultaneous Determination of the contents of 4 components of Qingwei Huanglian TabletsObjective: To establish a method for simultaneous determination of the contents of genpioside, baicalin berberine hydrochloride and ammonium glyeyrrhizinate of Qingwei Huanglian tablets.Methods: A good separation was achieved on a Hypersil C18 (250×4.6 mm, 5.0μm) column at room temperature. The mobile phases were consisted of acetontrile and 0.1% triethylamine (pH 3.0) with gradient elution at the flow rate 1.0 mL·min^-1. The detection wavelength was set at 230 nm.Results: The calibration curves were linear in the ranges of 4.82⁷7.2μg·mL^-1 for genpioside, 5.80⁹2.8μg·mL^-1 for baicalin, 1.63²6.1μg·mL^-1 for berberine hydrochloride and 6.40¹02μg·mL^-1 (r≥0.999 7) for ammonium glyeyrrhizinate, respectively. The average recoveries were not less than 98.0%.Conclusion: The method was convenient, rapid, and accurate. It was suitable for quality control of the production of Qingwei Huanglian tablets.(3) Determination of Genpioside, Baicalin and Berberine Hydrochloride in Gonglao Quhuo tablets simultaneouslyObjective: To establish a method for determination of genpioside, baicalin and berberine hydrochloride in Gonglao Quhuo tablets.Methods: The mobile phase composed of acetonitrile (A), acetonitrile-0.5% triethylamine (pH3.1) (10:90) (B). The flow rate was 1.0 mL·min^-1 with the wavelength at 254 nm.Results: The calibration curves were linear in the ranges of 3.22²06μg·mL^-1(r=0.999 9) for genpioside, 2.95¹86μg·mL^-1 (r=0.999 9) for baicalin and 1.64¹04μg·mL^-1(r=0.999 9) for berberine hydrochloride, respectively. The average recoveries were not less than 98.4%.Conclusion: The HPLC method, which was convenient, rapid, and accurate, could be used for quality control of production of Gonglao Quhuo tablets.PART 3 The Study of the In Vivo-In Vitro Correlation of Gonglao Quhuo Tablets in Rat(1) Study on Dissolution of Gonglao Quhuo tabletsObjective: The dissolution rate of Gonglao Quhuo tablets was determined, to provide basis for appraisal and control of Chinese medicine quality.Methods: According to appendix XCâ…¢of Chinese Pharmacopoeia 2005 edition Volâ…¡, dissolution medium was 500mL 0.5% SDS and rotating speed was 100 r·min^-1. The dissolution solution of tablet was taken and analyzed by HPLC method. A column of Kromasil C18 (4.6 mm×250 mm,5μm) was used. The mobile phase composed of A was acetonitrile, B were 0.5% triethylamine (pH 3.1) and acetonitrile (90:10), gradient eluation. The flow rate was 1.0 mL·min^-1 with the wavelength at 254 nm.Results: The average dissolution of 3 batches of this drug exceeded the amount by 75% in 3 h.Conclusion: An acute and qualitative method with good reproducibility to determine dissolution of Gonglao Quhuo tablets is established. (2) The Parmacokinetic Study of Bacalin and Berberine Hydrochloride in Gonglao Quhuo Tablets in RatsObjective: To establish an RP-HPLC method for deter- mination the content of baicalin and berberine hydrochloride in plasma and study pharmacokinetic of baicalin and berberine hydrochloride in rats in vivo.Methods: Plasma pretreated with methanol priciptation was performed chromatographic seperation. The concentration of baicalin and berberine hydrochloride in rat plasma was assayed after intragastic administration of Gonglao Quhuo tablets. A good separation was achieved on a Hypersil C18 (250×4.6 mm, 5.0μm) column at room temperature. The mobile phases were consisted of acetontrile and 0.1% triethylamine (pH 3.1) (30:70) at the flow rate 1.0 mL·min^-1. The detection wavelength was set at 265 nm.Results: The liner ranges of baicalin and berberine hydr- ochloride were 0.061³.91μg·mL^-1(r=0.999 7) and 0.017¹.06μg·mL^-1(r=0.999 9), respectivly. The lower limit of quantitation (LLOQ) of baicalin and berberine hydrochloride were 0.061μg·mL^-1 and 0.017μg·mL^-1, with accuracy and precision conformed to the requirement of biological specimen analysis. The distributions of bacailin and berberine hydrochloride in rats were conformed to be a two-compartment model and one-compartment model, respectively. The main pharmaco- kinetics parameters of baicalin were K₂₁= (0.740±0.30) 1/h, K₁₀= (0.143±0.035)1/h, K₁₂= (1.26±0.8) 1/h, t_1/2β= (13.5±1.0) h, t_1/2α= (0.399±0.18) h, T_max= (0.352±0.086) h, C_max= (0.940±0.18)μg·mL^-1, respectively. The important pharmacokinetics parameters of berberine hydrochloride were t_1/2(Ke) = (3.65±0.69) h, t_1/2(Ka) = (0.073±0.019) h, t1/2(Km) = (0.076±0.022) h, Tmax = (0.605±0.12) h, Cmax = (0.254±059)μg·mL^-1, respectively.Conclusion: The compatibility of complex prescription can affect some important pharmaco-kinetics parameters. The compatibility of complex prescription can affect some important pharmacokinetics parameters. A good correlation was achieved by evaluating the internal predictability of in vitro in vivo correlation.