Component Analysis And Multi-component Pharmacokinetics Of Fructus Aurantii

Objective:To identify the main chemical constituents in the various extracts of Fructus Aurantii(FA).To determine the blood compositions of FA by serum pharmacochemical study.Pharmacokinetic parameters of the main active components of FA were compared under different medicinal forms.It provides an analytical basis for the pharmacodynamic material basis and quality control of FA.Methods:1.UPLC-Q-TOF/MS method was used to characterize and identify the total components in FA,including methanol extract,water extract,and volatile oil,GC-MS was used to analyze the volatile oil,the characteristic components were analyzed by mass spectrometry.2.The theory of serum medicinal chemistry was guided and UPLC-Q-TOF/MS method was used to compare the differences FA extract,blank plasma,and drug-containing plasma spectra,then using Peakview and Metabolite Pilot data processing software,according to the retention time provided by mass spectrometry,accurate relative molecular mass,and cleavage fragments of MS/MS as indexes,combined with the results of previous studies to identify migrating components in rat plasma after oral administration.3.In this study,a rapid and efficient UPLC-MS/MS method was established for the simultaneous detection of the five active components(Meranzin hydrate,naringin,neohesperidin,meranzin,and nobiletin)in rat plasma,and the quercetin was used as an internal standard,Electrospray ionization(ESI)was used to switch between positive and negative ions,multiple reaction monitoring mode was used for quantitative determination,and further applied to the pharmacokinetic studies after rat oral administration of monomer,drugs in compatibility and FA extract.After hydrolysis with β-glucuronidase and sulfatase,the concentration of naringin and neohesperidin in rat plasma were expressed respectively by the total concentration of naringenin and hesperitin which was determined by UPLC-MS/MS,then we can analyze with DAS 3.1 software to calculate the pharmacokinetic parametersResults:1.In this paper,133 constituents were from systematically identified and inferred from the extract of Fructus Aurantii,which can be divided into 7 types of structural components,including flavanones,flavones,polymethoxyflavonoids,alkaloids,amino acids and peptides,limonoids and coumarins.94 volatile components were identified from the volatile oil of Fructus Aurantii2.After oral administration of the extract of the Fructus Aurantii,a total of 74 blood components were detected from the plasma,including 49 prototype components and 25 metabolites.Combined with the existing literature reports,it can speculated and identified that most of which were prototypes and metabolites of flavonoids,polymethoxyflavonoids,and small amounts of alkaloids3.UPLC-MS/MS method for simultaneous determination of Meranzin hydrate,naringin,neohesperidin,meranzin,and nobiletin in Fructus Aurantii.The five compounds are well separated and the response is high,the method has good linearity in the range of 0.2112～3880 ng/mL,the intra-day accuracy is between-8.66%to 9.68%,the intra-day accuracy is in the range of-9.34%to 9.94%,and the recovery rate is above 80%.There is no significant matrix effect,and the stability RE values are all-13.50%to 14.81%.The pharmacokinetic results showed that the mean plasma concentration-time curve of naringin and neohesperidin appeared double peaks,and were detected in rat plasma within 5 minutes after administration,and the peak Tmax was less than 1 h,Meranzin hydrate and meranzin were quickly absorbed(Tmax,about 1 h),but eliminated slowly(t1/2z over 6.5 h).According to Tmax and AUC(0-t),Nobiletin was also rapidly absorbed,with a Tmax value of 1.17 ± 0.26 h and an AUC(0-t)value of 472.64 ±118.70 μg/L*h.DAS 3.1 software was used to process the plasma concentration-time data of five components in rats.The optimal metabolic process was in accordance with the two-compartment pharmacokinetic model.Then further application of this method to study pharmacokinetics of monomers,compatible drugs and extracts of Fructus Aurantii after oral administration.The results showed that there were significant differences in pharmacokinetic parameters for naringenin and hesperetin between the compatibility,FA extract group vs monomer group:①remarkable increases in the values of AUC(0-∞),AUC(0-t)and Cmax;②obvious decrease of CLz/F;and③longer tmax and t1/2z.Conclusions:The main active components of FA include flavonoids,alkaloids and coumarins Among them,alkaloids,polymethoxylflavones and coumarin compounds are mainly introduced into the blood and play a role in the prototype,while the index components such as naringin and neohesperidin play an effect mainly through hydrolysis into aglycones.Naringin and neo-hesperidin can promote mutual absorption and influence metabolic behavior,and other components in FA extracts can synergistically promote the mutual absorption.To further analyze the pharmacodynamic material basis of FA,we need to integrate multidisciplinary methods and techniques,expand the research scope of the active components of FA,and systematically study the action process in vivo.