Quantification And Pharmacokinetics Study Of Bioactive Constituents From Zanthoxylum Armatum DC Using UFLC-MS/MS

Objective:Alkaloids and lignans,the main bioactive constituents,are widely distributed in the species of Zanthoxylum?Rutaceae?.It is reported the alkaloids,lignans,coumarins and volatile oils,etc.are the main bioactive ingredients of Zanthoxylum armatum DC.?Rutaceae?,and possesses many kinds of bioactivities,such as antinociceptive and anti-inflammatory,inhibit platelet aggregation,hepatoprotective,anthelmintic.In this study,a UFLC-MS/MS method was established to quantificate the alkaloids and lignans in the root,stem and branch of Z.armatum.And this is the first study of pharmacokinetics and tissue distribution study of bioactive constituents from Z.armatum in rat plasma and tissues.Methods:1.UFLC-MS/MS method was established to detected six alkaloids and four lignans?isodecaline?1?,6–acetonyldihydrochelerythrine?2?,N-Methylanhydrote-trahydroberberrubine A?3?,allocryptopine?4?,dictamnine?5?,escholidine perchlorate?6?,planispine A?7?,pinoresinol monomethyl ether?8?,de–4'–o–methylyangambin?9?and eudesmin?10??in Z.armatum.A Phenomenex Kinetex2.6u XB–C18 100A column?2.1×50 mm?was employed with the column temperature of 40?.The mobile phase consisted of water containing 0.1‰formic acid?A?and acetonitrile containing 0.1‰formic acid?B?with a gradient programmed as:0.00–0.50 min,2%B;0.51–5.00 min,40–75%B;5.01–6.00min,2%B.A 5?l supernatant was injected with a flow rate of 0.45 mL/min.2.A UFLC-MS/MS method was employed to determine the contents of three bioactive constituents from Z.armatum in rat plasma.This method was validated to detecting the contents of N-Methylanhydrotetrahydroberber-rubine A,dictamnine and eudesmin in rat plasma after orally administered with Z.armatum?1.4g/kg?extract.3.The contents of N-Methylanhydrotetr-ahydroberberrubine A,dictamnine and eudesmin in heart,liver,spleen,stomach,kidney,brain and caecum tissues of rat after orally administered with Z.armatum?1.4g/kg?extract were detected by UFLC-MS/MS.Results:1.The UFLC-MS/MS method was validated with precision,accuracy and stability.Extraction conditions were optimized by orthogonal design.Finally,the optimal ultrasonic extraction condition was 100:1 of methanol for 10min.The whole part of Z.armatum,from root to branch,was divided into 24 parts,then ultrasonic extracted.The results show that the contents of alkaloids and lignans were high in the root of Z.armatum,but lower in the middle and branch parts of the plant.2.The pharmacokinetic parameters of compounds show that the alkaloids,N-Methylanhydrotetrahydroberberrubine A and dictamnine,could quickly absorb into body,with T_max of 0.29h,and the T_max of eudesmin,a lignin,was longer than alkaloids'(T_max=2.5h).3.The results of tissue distribution show that the N-Methylanhydrotetr-ahydroberberrubine A,dictamnine and eudesmin were obviously enriched in stomach.Conclusion:1.The quantitative analysis result showed that the alkaloids and lignans were rich in the root of Z.armatum,but lack in the middle and branch parts of the plant.This result will be benifit to the quality control,further drug development and clinical application of Z.armatum.2.The pharmacokinetic studies of alkaloids and lignans showed that the alkaloids N-Methylanhydrotetr-ahydroberberrubine A and dictamnine reached C_maxax within 0.29 h,indicating that these two alkaloids can be rapidly absorbed into the body after oral administration.T_max of eudesmin was 2.5 h.And there was a double peak in the mean plasma concentration-time curve of N-Methylan-hydrotetrahydroberberrubine A,indicated a close relationship with enterohepati-ccirculation.3.The result of tissue distribution showed that N-Methylanhydrotetr-ahydroberberrubine A,dictamnine and eudesmin were enriched in the stomach.The content in stomach is significantly higher than other selected tissues,and this could relate to the traditional use of Z.armatum for the relief of stomach pain.And this could provide for clinical application.