Study On The Chemical Fingerprints Of Rhizoma Corydalis And Bulbus Fritillariae Thunbergii From Pan'an Province

Rhizoma Coridalis and Bulbus Fritillariae Thunbergii are two famous traditional Chinese medicines from Pan' an county of Zhejiang province, meanwhile they are included "eight sapor of Zhejiang". Because of the environment and climate there in Zhejiang province is special, the quality of these two traditional Chinese medicine is much better and much more steady than any other regions'. At present, some production bases in certain scale have formed. Though these medicine materials are partly from these production bases, most of them are purchased from farmers. Due to the nonstandard production, the quality is quite different, and different species may mix together. So it is necessary to identify the genuine medicine materials and build chemical fingerprints for them in order to guide good agriculture practice (GAP). This article builds High Performance Liquid Chromatography (HPLC) fingerprints and Fourier Transform Infrared Radiation (FTIR) fingerprints for Rhizome Coridalis and Bulbus Fritillaria thunbergii. At the same time, it analyzes the differences between genuine medicine materials grown in Pan' an and materials from other places, In the end, we build integrated chemical fingerprints of the two medicine materials from Zhejiang province.1. Fingerprints of genuine Corydalis yanhusuo W. T. WangThe traditional Chinese material Rhizoma Coridalis is the dry bulb of plant Corydalis yanhusuo W. T. Wang et Z. Y. Su et C.Y. Wu which belongs to family of Papaveraceae. It mainly grows in Zhejiang province and has been used to promote blood circulation, reinforce vital energy, and alleviate pain. The modern scientific study has proved that its major active components are alkaloids, especially tetrahydropalmatine which can alleviate pain effectively. In this article, the experiment used tetrahydropalmatine as control, then we compared different extract methods and different mobile phase systems. For the purpose of optimize the mobile phase systems; we added macromolecule organic acid, such as acetic acid and diethylamide to improve the peak shape and resolution. Finally, we ascertained the mobile phase systems and elution grade. Through the process of preparation, development analysis, evaluation and verification, we established the HPLC fingerprint of C. yanhusuo and compared the ones yielded from different places and different disposed methods. The consequence indicates that the fingerprint can reflect the resemblance of the medical material and provide reliable evidence to control the quality of C. yanhusuo. At the same time, we compared the distinction of the fingerprint of C. yanhusuo which yielded from different places and found that there is no significant difference in Zhejiang samples, but quite different between Zhejiang province and other province. The characteristic areas in the fingerprints of Pan'an areas are: 9min, 45min when there is a small peak in Pan'an samples but other samples don't have, and 70min, two samples of other provinces have an obvious peak, but Pan'an samples don't have. At the same time, we compare different disposed methods, the result showed: the fingerprint of the sample dealt with sulfur had less peaks than other samples, may be some componets lost or changed. The fresh materials and boiled ones have obvious differences which can identify from fingerprints and cluster analysis as well as PCA method also. Besides, we constructed FTIR spectrum fingerprints of C. yanhusuo yielded from difference places, and analyzed their similarity. The result indicated the similarity of the spectrum fingerprints is above 0.90, which is hard to be distinguished.2. Fingerprints of genuine Fritillaria thunbergii Miq.The traditional Chinese material Bulbus Fritillariae Thunbergii is the dry bulb of plant Fritillaria thunbergii Miq. which belongs to the family of Liliaceae. It mainly grows in Zhejiang province, Jiangsu province and An'hui province. Its active components are alkaloids, which are varied in different species. This article used the method from Pharmacopoeia of People's Republic of China (2005 Edition I) and made a little change which was using ultrasonic extraction method instead of distilling method recroded in Pharmacopoeia to extract their alkaloid components. We ascertained the mobile phase systems and elution grade through repeated process. Because most alkaloids have no ultraviolet absorption, we used Evaporative Light Scattering Detector (ELSD) to detect their components. We constructed five Fritillaria thunbergii Miq. fingerprints from Pan'an , one from Ningbo, one from Jiangsu and other three species. Through comparation, we ascertain the characteristic peaks of WanBei, they are: peak6, peak12, peak13, peakl5; the characteristic peaks of F. thunbergii Miq. var. chekiangensis Hsiao et K.O. Hsia are peak2, peak7, peak9, peak18; there are two samples from Sichuan province, they are belong to the same specie, just different in growth years, but the fingerprints of them are different, this might caused by the environment. Furthermore, we established the FTIR fingerprints of different Fritillaria species and chose the representational community associating with stechiometry, we can distinguish different species of Fritillaria by cluster analysis.