| Recently, adverse reaction and safety issues frequently occur during the clinical applications of TCMI due to their low quality standard and the unclear chemical material basis. Shenfu injection (SFI), a well known TCMI, is composed of red ginseng and black prepared lateral root of aconite, having rescuing from collapse by restoring Yang (阳) and tonifying Qi (气) to consolidate depletion effect, mainly used for JueTuo disease triggered by syndrome of sudden Yang (阳) collapse (such as infectious, blood loss, loss of liquid shock, etc.). In order to clarify the chemical components and improve the quality standard of SFI (a well-known TCMI), the chemical components of SFI and its raw materials were qualitatively and quantitatively characterized. The findings provided meaningful information to the understanding of the transformation and degradation pathways of chemical composition from raw materials to SFI and the basis for process parameters optimization and process control.1. HPLC-PDA fingerprint of Aconiti Lateralis Radix was developed by employing 12 batches of unprocessed materials (similarities 0.902-0.994), and 9 common peaks were observed. A total of 99 alkaloids were identified in unprocessed Aconiti Lateralis Radix using LC-MS-IT-TOF and 43 alkaloids from heishunpian were detected and tentatively identified using HPLC-MRM. Simultaneous determination of 12 alkaloids in unprocessed and processed Chuanwu, Caowu, and Wutou was conducted using HPLC-MRM. Results showed that more diester-diterpenoid alkaloids were distributed in the unprocessed materials, yet less diester-diterpenoid alkaloids in the processed os. Direct stability characterization of aconite alkaloids in different media by autosampler-mediated incubation-online solid phase extraction-LC-MS/MS. Extensive hydrolysis to generate two degradation products (dissociating the ester bond to lose the acetyl group at the C-8 position and expelling an acetic acid moiety from C-8 and C-15 positions) was observed for AC, MA, or HA in phosphate buffer saline (PBS, pH 7.4), PBS fortified with fresh rat plasma and PBS spiked denatured rat plasma, whereas BMA, BAC, and BHA remained intact in PBS, PBS spiked rat plasma, methanol, acetonitrile (ACN), and 50% aqueous ACN.2. As many as 221 ginsenosides were qualitatively analyze by hyphenating step-wise MRM and scheduled MRM modes, and putative identities were assigned to 197 ones. With the assistance of PCA analysis, biomarkers being responsible for rapid discrimination among ginseng and its different processed products were characterized. Quantitative determination of 11 ginsenosides (Rg1, Re, Rf, Rg2, Rh1, Ro, Rb1, Rc, Rb2, Rd, and Rg3) from unprocessed, processed ginseng was carried out using HPLC-ELSD. Results showed that ginsenosides of red ginseng had significant differences with others.3. A total of 157 hydrophilic components were detected from SFI, and 154 ones were annotated. Hydrophilic components were enriched by off-line preparing the 0.5 min-effluent of SFI. A pMRM-IDA-EPI strategy was adopted to characterize polar ingredients in SFI. Thirteen ginsenosides and 10 aconite alkaloids were quantified by online SPE-polarity switching LC-MS/MS. Ten amino acids,8 nucleotides,9 aconite alkaloids and 11 ginsenosides were quantitatively analyzed by column switching HILIC-RP Cis-LC-MS/MS. Four sugars (glucose, fructose, sucrose and maltose),15 organic acids,16 amino acids and 14 inorganic metal elements were quantified in SFI by HPLC-ELSD, HPLC-Qtrap-MS, amino acid analyzer, ICP-MS and ICP-OES, respectively.Above all, the analytical data in this study showed that SFI contained 0.002-0.029% aconite alkaloids,3.78-6.16% of ginsenosides,0.74-1.19% of the free amino acids,2.24-3.95% of the hydrolyzed amino acids,0.12-0.32% nucleotides,0.06-0.35% of organic acids, and 3.55-6.05% of inorganic metal elements. Chemical compositions of SFI were clarified by qualitative and quantitative characterized from raw materials to SFI, provided a basis for quality standard ascension of SFI and better ensured the clinical safety.Qishenkeli (QSKL), a well-known effective clinical experience formula, is usually utilized for nourishing Qi (气) and warming Yang (阳), invigorating blood circulation. Corresponding to the main etiology and pathogenesis coronary of heart disease and cardiac insufficiency, QSKL was prescriped on the basis of compound compatibility theory of monarch, minister, assistant, and guide. This formula consists of Astragali Radix, Salviae Miltiorrhizae Radix, Lonicerae Japonica Flos, Aconiti Lateralis Radix Preparata, etc.Aiming to clarify the effective material basis of QSKL, chemical database and fragmentation pathways of the main constituents were summarized on the basis of literature data. Firstly, structural identification was conducted for the primary chemical types in QSKL, and the original sources of the components in QSKL were completely assigned via comparing the LCMS-IT-TOF chemical prifles between QSKL and its constituent herbs. A total of 41 compounds were plausibly annotated in positive and 167 in negative. Secondly, metabolites in plasma, bile, urine, and feces of rats were characterized following the oral administration of QSKL by the high resolution mass spectrometry (LCMS-IT-TOF). A total of 260 compounds were plausibly annotated in positive, including 43 prototypes and metabolites in plasma,71 prototypes and metabolites in bile,68 prototypes and metabolites in urine and 78 prototypes and metabolites in feces, and 411 compounds were plausibly annotated in negative, including 52 in plasma,134 in bile,93 in urine and 132 in feces, respectively, by LCMS-IT-TOF combined fragmentation rules of chemical components, diagnostic fragment ions-based filter or comparison with retention time of reference substances. Deglycosylation, oxidation, hydrogenation, sulfonation and glucuronidation were observed as the primary metabolic pathways for the chemical constituents of QSKL. Thirdly, pharmacokinetic profiles of 25 compounds were studied in rat after intragastric administration of QSKL, and concentration-time curves and pharmacokinetic parameters were obtained for 21 compounds. After comparing the model group with control group, glycyrrhizic acid, glycyrrhetinic acid and tanshinone ⅡA were presumably considered to contribute to its therapeutic effects on coronary heart disease and cardiac insufficiency.Preliminary data illustrated the pharmacokinetic characteristics of constituents migrating to blood, providing reference for QSKL efficacy material basis research, action mechanism and clinical use. |
PDF Full Text Request