Quality Assessment Of D. Genkwa Its Processed Products

Purpose: To develop qualitative and quantitative evaluation methods, fully understand the chemical constituents and study theory basis of processing so as to normalize quality standard of Flos Genkwa. The preparation technics of flavonoid genins has also been studied. Above-mentioned tasks are expected to be propitious for quality control and further developping of the drug.Materials and Methods:Material: Flos Genkwa ( 23 batches ), Flos Chamaedaphne ( 1 batches ). Reference Substances: genkwain and yuanhuacine (Chinese medicine institute of China academy of TCM, luteolin (China pharmaceutical and biological products institute), apigenin (Kunming Kexiang Biological Technology Co., LTD.). Chromatography pure methanol (Tianjin Kemiou), chromatography pure acetonitrile (Tianjin Xiehe), Wahaha pure water. All other reagents are analysis pure.Methods: Do qualitative analysis by TLC, HPLC fingerprints and LC-MS. Do quantitative analysis by HPLC and UV-vis. Investigate the preparation technics of flavonoid genins through single factor text and orthogonal text.Results:1. Flos Genkwa and Flos Chamaedaphne were qualitatively identified by TLC. The method can distinguish the two varieties with good separation and moderate Rf.2. The fingerprint atlas of flavonoid ingredients were established by determining 13 batches of Flos Genkwa, and 16 peaks were selected as characteristic peaks. According to the results of cluster analysis the fingerprint was divided into two regions:Ⅰfinger region for identification,Ⅱmeasurement region for quality evaluation.3. Extract of Flos Genkwa was analyzed by LC-MS, and 15 ingredients were identified.4. Genkwain and yuanhuacine were determined simultaneously by HPLC with changing ultraviolet-visible wavelength. The average recoveries were 97.7% and 97.1% and RSD were 2.28% and 1.64% (n=5). The method appears to be convenient, reliable and repeatable. Five batches of crude drugs were determined, as well as ten batches of pilot pieces, and the latter verifies stability of the processing method.5.Three methods were used for analysis of total flavonoids in Flos Genkwa. Determined by hydrochloric acid - magnesium colouration can satisfy the analysis requirement, and the method is reliable and repeatable.6. Comparation research to four kinds of processing methods was done. The results showed that: processing with vinegar is indeed able to reduce the toxic components ----yuanhuacine.7. Optimized preparation technics of flavonoid genins was got through single factor text and orthogonal text. 100℃, 2% acid hydrolysis, 2 hours is the best choice.Conclusions:1. TLC identification method was optimized. HPLC method for fingerprint is simple, accurate, and seneitive. Flos Genkwa and its counterfeit can be distinguished intuitively from the fingerprints, also the atlas can provide information of genetic distances and quality. LC-MS analysis provides us a comprehensive understanding of the effective components and references for further research.2. The methods of simultaneous determination of genkwain and yuanhuacine and determination of total flavonoids are both simple, reliable and repeatability, and can be used to control the quality of the drug.3. Processing with vinegar can reduce the toxicity of the drug. Vinegar is the most important factor.4. Hydrolyzing changes glucoside to genin, which reduces the stimulation of the drug and makes active ingredients concentrated. The study verified the reasonability of boiling with vinegar from the side, namely reducing toxicity and improving effectiveness.