The Study On Chemical Fingerprint And Metabolic Profile Of Non-volatile Components Of Herb Pair Citri Reticulatae Pericarpium-Pogostemonis Herba

Herb pair refers to the clinical relatively fixed composition of two herbs is the most common, simplest and most basic form of traditional Chinese medicine(TCM) compatibility. Herb pair is two herbs combination which are commonly used in clinical medicine for the relative fixation. Citri Reticulatae Pericarpium, Chinese means Chenpi, is Citrus reticulata Blanco and its cultivars of dry peel. Pogostemonis Herba, Chinese means Guanghuoxiang, is the dry ground part of Pogostemon cablin(Blanco) Benth. They are two common Chinese herbal medicines, compatibility are widely used. The compatibility of them was first recorded in the Chenpi-Huoxiang Decoction which from the sixth volume of “Yixue Congzong Lu” for treating injury summer irritable, vomiting and diarrhea and cholera.According to ChP. 2015, there are 24 prescription preparations for this herb pair’s application. However, there are no reports of non-volatile components of Herb Pair Citri Reticulatae Pericarpium- Pogostemonis Herba in the existing literature data. Thus, in this paper, we apply UPLC-DAD-Q/TOF-MS technology, to study its non-volatile components of the herb pair. For the purpose of making full use of the medicinal herb, several parts from herb pair are studied.First, UPLC-DAD-Q/TOF-MS were applied to simultaneously characterize the non-volatile components of herb pair Citri Reticulatae Pericarpium- Pogostemonis Herba. Analysis was performed on an Endeavorsil C18, Rapid Resolution column(2.1 mm×50 mm, 1.8 μm, Dikma) with gradient elution of methanol(A)-aqueous solution containing 0.2% formic acid(B) at a flow rate of 0.3 mL/min and the column temperature was 25 ℃. By comparing MS fragmentation, accurate molecular weight, literature date and standard compounds information, a total of 18 compounds were successfully identified or speculated. The origins of these compounds were assigned to the corresponding Chinese medicine. UPLC-DAD-Q/TOF-MS is a simple and rapid tool for the identification of the non-volatile components of Herb Pair Citri Reticulatae Pericarpium- Pogostemonis Herba, and the results could provide evidence for the research on quality combined and effective constituents of Herb Pair Citri Reticulatae Pericarpium- Pogostemonis Herba.Second, using the technology of UPLC-DAD-Q/TOF-MS, we analyze the extract of the plasma metabolic which are collected from SD rats after oral non-volatile components of the extract from herb pair Citri Reticulatae Pericarpium-Pogostemonis Herba. Analysis was performed on an Endeavorsil C18, Rapid Resolution column(2.1 mm×50 mm, 1.8 μm, Dikma) with gradient elution of methanol(A)-aqueous solution containing 0.2% formic acid(B) at a flow rate of 0.3 mL/min and the column temperature was 25 ℃. The plasma metabolic fingerprint spectrum has carried on the preliminary analysis and detection of bleeding may exist in the plasma of six prototype compounds and 20 related metabolites. The mass spectrometry of the prototype and metabolic product of combined with related literatures, we found that the 20 metabolites were the biotransformation of the six prototype compound, and biotransformation progress mainly includes methyl reaction, glucuronidation and sulfation reaction. This study provides reference method and experimental basis for metabolism research and clinical application of Herb Pair Citri Reticulatae Pericarpium- Pogostemonis Herba.Last, Ultra High Performance Liquid Chromatographic(UPLC) method was developed for the simultaneous determination of Vicenin-2, Hesperidin, Nobiletin and Tangeretin in Chenpi-Huoxiang Decoction. The separation was performed on a Endeavorsil C18 column with methanol and 0.2% formic acid as the mobile phase of gradient elution at a flow rate of 0.3 mL/min. The detection wavelength was set at 300 nm. According to the result, the linear range of Vicenin-2 was 4.26~12.6 μg/mL(r=0.9992). The average recovery for Vicenin-2 was 97.4%,precision of the method was 2.38%(RSD,n=6). The linear range of Hesperidin was 9.52~78.4 μg/m L(r=0.9999). The average recovery for Hesperidin was 98.9%,precision of the method was 1.09%(RSD,n=6). The linear range of Nobiletin was 4.00~12.0 μg/mL(r=0.9997). The average recovery for Nobiletin was 100.3%,precision of the method was 1.71%(RSD,n=6). The linear range of Tangeretin was 1.76~3.96 μg/m L(r=0.9993). The average recovery for Tangeretin was 103.5%,precision of the method was 1.22%(RSD,n=6). The method is simple,accurate and reproducible. It is a good method for the quality control of Chenpi-Huoxiang Decoction.The chemical fingerprint in vitro and metabolic profile in vivo of non-volatile components of herb pair Citri Reticulatae Pericarpium- Pogostemonis Herba were established in our study. Meanwhile, we established a suitable method for determination of 4 component of Chenpi-Huoxiang Decoction. We have provided the useful expermental result for the material basis of the Herb Pair Citri Reticulatae Pericarpium- Pogostemonis Herba, and supplied experimental basis for the follow-up study of Citri Reticulatae Pericarpium- Pogostemonis Herba, and also promote containing the herb pair for further research in other preparation.