Evaluation Of The Quality Of Suphora Flavercens Ait.in Guizhou

Objective: To study the relationship between ecological environment factors and determination of matrine in Suphora flavercens Ait. and analysis of heredity and variation by the research of pharmacognosy. These provide a basis for quality evaluation and artificial cultivation. Method: The research of pharmacognosy include the original plant identification, morphological identification, routine examination of medicinal materials, microscopical identification and physical and chemical identification and DNA molecular genetic maker. The microscopical identification used free-hand section and the process of chloral hydrate. Ultrasonic technology was used to extract matrine. these measures, which was used to quantity and quality analysis, ultrasound technology, uv-spectrophotometric and HPLC.modified CTAB method was used to extract the genomic DNA of Suphora flavercens Ait. The suitable ISSR-PCR reaction system was establish by orthogonal design L16(45). The optimal conditions in 25 μL ISSR-PCR system were 90 ng DNA template, 0.4 μmoL·L-1 Primer, 2.0 mmoL·L-1Mg2+, 0.5 U Taq DNA Polymerase, 0.5mmoL·L-1 dNTPs. The optimized reaction system by orthogonal design is suitable for ISSR-PCR analysis of Suphora flavercens Ait., which provide the base to analyze genetic diversity of Suphora flavercens Ait. Clustering analysis was carried out on the different origin of polymerase chain reaction bands. Results: The microscopic structure of the stem is same as the published report. Determination of matrine make a big difference. Altitude and average annual rainfall have a significant influence to determination of sophocarpidine. Average annual temperation and annual average sunshine hours have no significant influence to it. Matrine, which is 0.015 μg~0.12 μg,has a linear relationship with peak area( r=0.9995). Oxymatrine, which is 0.0585μg~0.8395 μg, has a linear relationship with peak area( r=0.9990). The average sample recovery rate of matrine was 96.67% and RSD=1.65%( n=9). The average sample recovery rate of oxymatrine was 98.46% and RSD=1.13%( n=9). DNA extraction kit is better than improved CTAB. The optimal conditions in 25μLISSR-PCR system were 90 ng DNA template, 0.4 μmoL·L-1 Primer, 2.0mmoL·L-1Mg2+, 0.5 U Taq DNA Polymerase, 0.5 mmoL·L-1 dNTPs. The matrine from 16 origin were divided into three classes and whose similarity coefficient was0.00 ~ 0.49. Conclusion: Altitude and average annual rainfall have a significant influence to determination of sophocarpidine. Average annual temperation and annual average sunshine hours have no significant influence to it. This provides experimental basis for artificial cultivation technology of Suphora flavercens Ait. These samples meet the standard of the 2010 edition of the Chinese Pharmacopoeia except Bijie County, Jinyang Century City, Liping County, Qiandongnan province. DNA extraction kit is better than improved CTAB. The optimized reaction system by orthogonal design is suitable for ISSR-PCR analysis of Suphora flavercens Ait.,which provide the base to analyze genetic diversity of Suphora flavercens Ait.