Studies On The Chemical Figerprinting Of Qingkailing Injection

It is well known that the quality control (QC) of traditional Chinese medicine (TCM) is one of the key tasks of TCM modernization. Chemical fingerprinting of TCM, which is already internationally accepted as an efficient QC technique, is an integral expression of the consitutents in TCM and thus can properly express their properties of complexity and interdependence. In this study, focused on the problem of QC, systemic studies on the fingerprinting and its application of Qingkailing injection (QKL) were carried out. Novel techniques of multi-marker components quantitative fingerprinting and two-dimensional fingerprinting were further developed, which could be applied as new techniques for the QC of traditional Chinese medicinal injection.The main contents of the dissertation are as follows:(1) Fingerprinting of raw medicinal materials of QKL were studied. QC of medicinal materials is considered as the foundation of that of QKL. Fingerprints of three most important materials of Radix Isatidis, Flos Lonicerae, Fructus Gardenise were studied, from which, quality evaluation were then successfully conducted on various medicinal materials from various QKL manusfacturers. In addition, focused on the volatile constituents of Fructus Gardenise, a new sampling method of directly sample introduction (DSI) was employed to establish the DSI/GC/MS fingerprint, which was a rapid and cheap solution for the fast quality evaluation of medicinal materials contained volatile constituents.(2) Chemical fingerprinting of QKL was studyed. The fingerprint was established to express the chemical information mainly on the chemicals derived from materials of Radix Isatidis, Flos Lonicerae, Fructus Gardenise, hydrolysate soluation (hydrolysate of Cornu Bubal and Concha Margaritifera). Fringerprints of QKL and its medicinal materials were compared and the origination study of the main chromatographic peaks was performed. Furthermore, since steroids having poor UV absorptions, detection technique of evaporated light scatter detection (ELSD) was employed to establish HPLC/ELSD fingerprint of QKL, which was applied to the quality evaluation of 19 QKL samples from different manufacturers and preparation procedures by constellation clustering method.(3) In this study, it was proposed that QC should be conducted base on the effectual segments of TCM, and multi-marker components quantitative fingerprinting was further developed. There are five major "effectual segments" in QKL, say nucleoside, iridoid glycoside, organic acid, chlorogenic acid, flavone glycoside and steroid. In according to the special chemical and spectral characteristics of these components, ELSD and the combined detection techniques of DAD/ELSD were employed, from which, two multi-marker components quantitative fingerprints of HPLC/DAD/ELSD and HPLC/ELSD were established to simultaneously determine five structural types of compounds. The proposed multi-marker components quantitative fingerprints were applied to evaluate the quality of QKL samples of different manufacturers and satisfactory results were obtained.(4) In order to overcome the shortcomings of conventional HPLC/UV fingerprinting, a novelapproach of two-dimensional (2D) fingerprinting was proposed for the first time. 2D fingerprint was generated by principal component analysis of DAD chromatographic data, which was, in comparison with conventional HPLCAJV fingerprint, more informative since two-dimensional information of spectra and chromatogram were combined. 2D fingerprint was applied to the quality evaluation of QKL samples from different manufacturers and preparation procedures, and it was found to be a more powerful tool to characterize the quality of TCM products.(5) Chemical fingerprinting was applied to control the procedure conditions in the production of QKL. Samples were collected from production spots of QKL at various procedures. HPLC/UV fingerprint of each sample was obtained according to the developed method, from which, the changes of constituents could be investigated, and then the diagnosis and optimization of procedure conditions were performed. The proposed fingerprint-based procedure controlling technique was employed by several manufacturers, and as a result, the quality of the products of each manufacturer was improved obviously.A comprehensive quality control approach of QKL based on chemical fingerprinting was developed according to the above works. Techniques of multi-marker components quantitative fingerprint and 2D fingerprint were further proposed, which enriched and promoted the present fingerprint-based analysis theory, and provided new solutions for the quality control and procedure control of TCM.