LC-HR-MS ~ N Of Kudiezi Chemical Constituents Of Injections And Xueshuantong Based

Kudiezi and Xueshuantong injections have been playing an important role in treatment of cardiovascular and cerebrovascular diseases, which are applied widely in clinic. However, it is not clear about their effective constituents, owing to the low isolation and sensitivity of traditional separation methods. However, LC-MS technology has combined the excellent separation ability, high sensitivity and selectivity for complex samples. Moreover, various hybrid mass spectrometers have been applicated widely. It can facilitate to obtain rich structures and fragmentation information of compounds, which are contribute to high efficiency and accuracy of identification. Moreover, it is propitious to the detection of macro-and micro-constituents simultaneously. Mass defect filter technology based on high-resolution mass spectrometry data could facilitate to compound screening. Therefore, focusing on the sensitivity of detection, trigger of multistage dissociation and data post-processing, compounds in Kudiezi and Xueshuantong injections were studied with HPLC-LTQ-Orbitrap technology and MDF methodology. The main research contents and results were summarized as follows:1. Identification of constituents in Kudiezi injectionDue to the chromatographic coelute and ion suppress during the course of LC-MS detection, the MS/MS fragmentation reaction of microconstituents were difficult to be triggered, which indicated that inadequate information could be obtained to elucidate the structures. Hence, full scan-dynamic exclision (FS-DE) and multiple ions monitoring (MIM) were applied to detect phenolic acids, flavoniods, and sesquiterpene lactones in Kudiezi injection utilizing HPLC-LTQ-Orbitrap mass spectrometry. For FS-DE, first, compounds within the setting range were detected coupled to dynamic exclusion and data dependent scan. Multi-stage mass information could be acquired automatically. FS-DE is adopted to enhance the detection efficiency and throughput. Target ions could be detected utilizing MIM. Moreover, MS/MS reactions of macro-and micro-consitituents could be triggered coupled with DDS. MIM is adopted to enhance the detection sensitivity, signal to noise ratio and specificity of mass information, eliminate interference, and facilitate identification of compounds in complex matrics. According to the high resolution information data, molecular formulae of compounds were predicted. Combined the characteristic of fragmentation pathway and multiple fragmentation data,25phenolic acids,28flavonoids, and50sesquiterpene lactones were screened and identified. This work was greatly contribute to the study of PK-PD. The two high-resolution detection methods with high sensitivity could be employed as rapid and accurate technique to screen and indentify compounds from TCMs.2. Screening and identification of polyphenols in Kudiezi injection and metabolites of flavonoids in rat plasma base on mass defect filter data-processing technologyAccording to characteristics consitituents in traditional Chinese medicine and bio-transformaton in vivo, the strategies have been developed for screening and identification of polyohenols in Kudiezi injection and metabolites of flavonoids in rat plasma, utilizing high resolution mass spectrometer and mass defect fiter technology. First, it is necessary to define parent nucleus and substituents for a certain kind of consitituents to determine the corresponding mass range and mass defect range. Second, multiple mass defect filters (MMDF and Stepwise MDF) were adopted to process the HR-MS data based on the subsitituents and interference strength of background matric ions, which could enhance the detection rate of compounds. Last, fragmentation regularities were used to rapidly identify the candidates screened and structure identification. As a result,30polyphenols in Kudiezi injection and18metabolites in rat plasma were exposed. The results indicated that the MDF could be employed as a rapid, effective technique to screen and identify compounds from TCMs.3. Identification of saponins in Xueshuantong injectionVarious of saponins in Xueshuantong injection were contained. Then the saponins in Xueshuantong injection were detected by FS-DE based on HPLC-LTQ-Orbitrap. According to the high resolution data and fragments, types and number of sugar moiety of saponins were deduced. Compared with literatures,78saponins were identified. The results indicated that FS-DE detection method could improve the throughput and be employed as an effective technique to screen and identify macro-and micro-constituents in TCMs.