| This paper focused on the study of phytochrome in Qingkailing injection systemat ically according to using the combination of UV-VIS, HPLC-MS/MS". Through the scr eening of crude drug, Fructus Gardenia, Flos lonicerae and Radix Isatidis could lead to the injection of the colored medicinal-Fructus Gardenia, Flos lonicerae and Radix Isatidis carries on the analysis, Finally this experiment choose Fructus Gardenia and Flos lonicerae for research. Finally the research aimed at two chief chemical compone nts-crocin and flavonoid, and the made quantitative analysis for multiingredient. It co uld play an important part for adverse reaction resulting from Qingkailing injection an d prepare for solving the clear level problem of Chinese Medicine injection.This paper studies the specific content of include the following three parts:1. The identification of the pigment components in Fructus Gardenia The research of crocetin componentsHigh performance liquid chromatography (HPLC)-photodiode array detector (DAD)-mass spectrometry (MS) was used to build the fingerprint of crocin in Fructus Garde nia. In t-he positive and negative mode, the components of crocin were studied in det ail. Grasp-ing the relation of the structure of such compounds and cracking behavior i n mass sp-ectrometry, and summarizing fragmentation pathways about this kind of co mpound in electrospray ionization and negative ion model, we could test and verify it according t-o its ultraviolet spectrum characteristics. The separation was perfoemed on a Dikma Diamonsil C18column (4.6mm x250mm,5μm); Mobile phase for0.3%acetic acid-acetonitrile gradient elution; Flow:1mL·min-1, diversion ratio for4:1; colu mn temperature:30℃; Photodiode array detector:190～700nm uv-vis spectra, detecte d at440n-m; Mass spectrometry conditions:ESI ion source:negative mode; Dry gas11L·min-1;Spray chamber pressure:50psi; Atomization temperature:350℃; Capillary voltage:35-00eV; Scanning range m/z100～1300; Mobile phase after the detector shunt into d-etector for0.25mL·min-1. Finally we could get ultraviolet spectrum data, precise mol-ecular weight and multi-stage mass spectrum data analysis about34kinds of crocins.Based on the inference to the standard of multistage mass spectrometry an alysis of thelaw that cracking, and literature data control, We could identify structure of26kinds of crocetin.The determination of components of crocinThe fingerprint of components of crocin in Fructus Gardenia was established by using hig-h performance liquid chromatography (HPLC) and make fast identification. T he prima-ry components-crocetin I and II crocin were determined.Flavonoid ingredientsAccording to analysis of the existing flavonoids standard product by HPLC-DAD-MS/MS and mass spectrometry of the standard of the product with debris literature m-aterial, we summarized the class of compounds of mass spectrometry cracking charact-eristics. Finally structures of six compounds were identified by a detailed analysis of the research for the Fructus Gardenia flavonoids ingredients pigment in the positive a nd negate-ve mode.2. The identification of the flavonoids components in Flos loniceraeSummarizes the mass spectrometry cracking characteristics of the flavonoids by an-alyzing the fragments of the standards using HPLC-DAD-MS/MS and integrating with references. The focus of the structure analysis of these compounds is the identification of flavone aglycone and glycosyl, including the identification of connection structure o-f glycosyl and aglycone. Most of the sugar in flavonoid glycoside is known and the kinds of sugar are not much. Structure determination of the sugar chains is usually ba-sed on the identification of known saccharide grous. So the key point is to identify t-he position of glycosidation and the configuration of glycoside. It is an effective way to analyze the structure type of these compounds using UV spectra. This study could identify the structure of the compounds quickly and effectively by MS data, UV spec-trum, retention time, and so on. Meanwhile, analytical data of the standards and datas upplied by references are helpful. Sometimes, this method could find new compounds.In this study, we analyze the Flavonoid pigments in Flos lonicerae in detail by p ositive and negative mode. The separation was perfoemed on a Dikma Diamonsil C18column chromatography (4.6mm x250mm,5μm); Mobile phase for0.3%acid solu tion-acetonitrile gradient elution; Velocity1mL·min-1. diversion ratio for4:1; Thecolu mn temperature30℃; Photodiode array detector record190-400nm ultraviolet spe ctrum, detected wavelength set for355nm;Mass spectrometry conditions for:ESI ion source anion mode; Dry gas velocity and L·min-1; Spray chamber pressure50psi; At omization temperature350℃; Capillary voltage3500eV; Scanning range m/z100～1400; Mobile phase after the detector shunt into detector for0.25mL·min-1. Finall-y, we got18flavonoid data and identified4compounds’ structure. Also, we inferred13compounds, basic structure of Flos lonicerae.3. Analysis of the components of pigment in Qingkailing injectionAccording to the established testing method of the detection to the components of crocins and flavonoids and comprehensive analysis of little content of the two components in Qingkailing injection, three kinds of the components of crocin and the structure of two flavonoids were inferred. At the same time two kinds of the components of crocin and the structure of two flavonoids were confirmed. In addition, this could supply a comprehensive analysis for the study of pigments in Qingkailing injection.A simple and fast analysis and identification of the mass spectrometry pigment composition analysis methods were established through the research, and these methods were regarded as the judgement of pigments in Qingkailing injection. This method could not only analyze fingerprint of regular pigment of the original medicinal materials, but also make quantitive analysis to the original medicinal materials of the injections, and provides references to the envaluation of the traditional Chinese medicine injection. |
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