Study On The Active Constituents In "Xue-Fu-Zhu-Yu Capsule" And Quantitative Analysis Of Steroid Saponins In Rhizoma Paridis

In this study, chemical constituents in extract and rat plasma were studied to searchfor the active constituents in XFZY capsule. Firstly, LC-MS technique was applied to identify the chemical constituents in XFZY capsule, Fructus Aurantii and Semen Persicae, respectively; then constituents absorbed into blood were detected to determine the active constituents. The detailed results were as follows:1. A rapid, sensitive and selective ultra-performance liquid chromatography-photodiode array detector - quadrupole time-of-flight mass spectrometry(UPLC-PDA-Q-TOF/MS) method was developed for analysis and identification of the bioactive components and their metabolites in rat plasma following oral administration the aqueous extract of F. aurantii. Twelve flavonoid O-glycosides were detected, and nine compounds were tentatively identified as polymethoxylated flavones. Six parent compounds were identified and four metabolites were observed in rat plasma, two of which were identified as naringenin glucuronide and hesperetin glucuronide, respectively. The results indicated that naringin and neohesperidin may be the active compounds in F. aurantii, which works as parent compound and its metabolite.2. An HPLC method for determinning naringin and neohesperidin in rat plasma was established, and the pharmacokinetic parameters can clarity the pharmacokineticsprocess of F. aurantii in rat.3. An UPLC-PDA-Q-TOF/MS method was developed to analyze the aqueous extract in S. persicae and the constituents absorbed into blood for the first time. Eleven compounds were identified, including two flavonoid O-glycosides, six cyanogenetic glycosides, threonine, chlorogenic acid and (+)-catechin. Six compounds were detected in rat plasma, including two parent compounds amygdalinand prunasin, two metabolites of amygdalin. The results indicated that cyanogenetic glycosides especially amygdalin are the active constituents in S. persicae.4. An LC-MS method was developed to analyze the active constituents in XFZY capsule for the first time. Twenty four major constituents from XFZY sample, including eight flavones and flavonoid O-glycosides, three terpenoid saponins, nine phthalides, one cyanogenetic glycoside, one chalcone, one organic acid and one nucleotide, were identified or tentatively characterized based on their retention times, DAD and MS data. Furthermore, compounds such as amygdalin, naringin, neohesperidin, naringenin glucuronide and hesperetin glucuronide were detected in rat plasma.Current quality control patterns are limited to industrial application, for most the natural chemical reference substances are expensive and unavailable. Here in, quantitative analysis of multi-components with single marker (QAMS) method, is established and validated to simultaneously determine five steroidal saponins (Paris-VII, Paris-H, Paris-II, Dioscin , Paris-I) in Rhizoma Paridis. Using Paris-I as the contrast, the relative correction factors (RCF) of the other four steroidal saponins are determined by HPLC-UV. The RCF has a good reproducibility in various instruments, chromatographic columns (RSD=0.88~4.52%). According to their RCF, five teroidal saponins are simultaneously determined in Rhizoma Paridis by one marker.