| Chinese herbal preparations are not only to prevent and treat disease but also to provide wide resources for exploitation of new drugs. Therefore they have become increasingly popular. In order to using Chinese herbal preparations rationally and safely, it is necessary to build sensitive and accurate analytical methods for deepening the study of pharmacokinetic, pharmacology, toxicology and qualitative standard. Chinese herbal preparations are not as simple and standard as chemical drugs, they have complex components which affect and act each other, in which the analytical result of a compound is probable affected by change of parent herb or prescription. It is necessary to build different analytical methods for a compound in Chinese herbal preparations according to analyzed sample, purpose and requirement. The HPLC is convenient to operate with high effectiveness, sensitivity and accuracy, so it is very suitable for the analysis of Chinese herbal preparations that have complex component and low content.On the basis of the previous literatures, the following major innovative works were carried out in this dissertation:(1) The novel HPLC methods were developed for the determination of chlorogenic acid and baicalin in both Yin-Huang preparations and rat plasma.(2) The pharmacokinetic study of Yin-Huang preparations in rat was carried outby HPLC, and pharmacokinetic parameters were calculated and discussed.(3) It was first to research the influence of administration dose and way to trendchange of chlorogenic acid and baicalin in rat dosed with Yin-Huang preparations.(4) A novel HPLC method was developed for simultaneous determination ofchlorogenic acid, caffeic acid, ferulic acid, protocatechuic acid andprotocatechuic aldehyde The developed method can be used for analysis and study of Fufang-Pugongying-Mixture and its compositional herbs.(5) The influence of preparation craft to the component content of Fufang-Pugongying-Mixture was studied by the developed HPLC method for the first time.(6) The study of the in vitro permeability of usnic acid, an effective component of Chinese herbal preparation was carried out through rabbit skin.This dissertation consists of six chapters.Chapter 1: In this chapter, the current status of HPLC technique is given by complete description at development of HPLC, method of qualitative analysis, use of union technique and the application of HPLC in study of Chinese herbal preparations.Chapter 2: A novel RP-HPLC-UV method for the quantification of CGA (chlorogenic acid) in rat plasma was developed. A good linear relationship between CGA concentration in plasma and corresponding peak-area ratio (CGA / internal standard) was obtained in the concentration range of 0.4—40 |ag/ml. The recovery of CGA was in the range of 85.29—89.12%, and LOQ (the limit of quantification) was 0.4 (xg/ml. The analytical method has high precision (RSD < 10.78%) and accuracy (-2.20% < RE < 5.00%). The method was successfully applied to the pharmacokinetic study of CGA in rat following administration of Yin- Huang granule, and the influence of administration dose and way to trend change of CGA in rat blood was researched.Chapter 3: A novel RP-HPLC-UV method for the quantification of BCL (baicalin) in rat plasma was developed. A good linear relationship between BCL concentration in plasma and corresponding peak-area ratio (BCL / internal standard) was obtained in the concentration range of 0.3—30 ng/ml. The recovery of BCL was within 96.31— 107.22%, and the LOQ was 0.3 ug/ml. The analytical method has high precision (RSD<8.08%) and accuracy (-3.00% < RE < 6.67%). The method was successfully applied to the pharmacokinetic study of BCL in rat following administration of Yin-Huang granule or Yin-Huang oral liquid, and the influence of administration way totrend change of BCL in rat was researched.Chapter 4: A novel RP-HPLC-UV method was developed for simultaneous determination of chlorogenic acid, caffeic acid, ferulic acid, protocatechuic acid and protocatechuic aldehyde. The good linear relationships between concentrations and corresponding peak-areas were obtained in the concentration range of 2—200 ng/ml for the 5 compounds mentioned above. The recoveries of(the 5 compounds were in range of 96.72—104.07%. Their LOD (the limit of determination) values were in range of 0.05—0.78 {ig/ml, and the CV (the coefficient of variation) values reflecting analytical precision in range of 0.89—4.5%. The method can be used as qualitative method of Fufang-Pugongying-Mixture (FFPM), and used for the study of preparation craft of FFPM and analysis of Taraxacum, Lonicera and Angelica.Chapter 5: Through evaluation and comparison of analytical method, the RP-HPLC-UV methods for determining respectively CGA and BCL were selected and developed. The good linear relationships between concentrations and corresponding peak-areas were obtained in the concentration range of 1—50 fxg/ml for CGA and BCL. The LOQ of CGA was 0.2 (j.g/ml and BCL was 0.3 ug/ml, and their recoveries were between 97% and 104%. The analytical methods have high precision and accuracy: RSD<5%, -3% |
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