The Content Analysis And Pharmacokinetic Study Of Six Aconitum Alkaloids Of Fuzi Based On UPLC-MS/MS

Objective:The research is trying to develop an analyzes methods of six aconitum alkaloids in plasma to study the toxic Chinese medicine Fuzi and discuss the pharmacokinetic process of aconitum alkaloids after taken by beagle dogs. We aim to figure out the metabolic rule of Fuzi.Methods:(1) UPLC-MS/MS method was used to establish the method of determine six aconite decoction in Fuzi, water decoction of Fuzi and its compound prescription to analyse content of Fuzi and its water decoction.(2) Sample preparation was performed with solid-phase extraction(SPE) on a3mL HLB cartridge before the analysis and Hydrobromide was used as the internal standard. UPLC-MS/MS method was used to establish the method of determine six aconite decoction in plasma.(3) An ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for simultaneous quantitation at different time of six Aconitum alkaloids in beagle dog plasma after oral administration of Fuzi decoction. The pharmacokinetic parameters were calculated by the results of the analysis through the DAS2.1.1software (Drug and Statistics for Windows).Results:(1)The UPLC-MS/MS method for simultaneous determination of AC, MA, HA, BAC, BMA, BHA, had a good separation degree whit high response. The linear regression for alkaloid was obtained in the range of0.5-200ng/mL. The LLOQ of the method was0. lng/mL and the LOD is0.05ng/mL. The recovery rate was over97%, and the analysis time was only taken7minutes. The alkaloid content of Fuzi provided by four hospitals was meted a criterion of pharmacopoeia. The aconite alkaloids content of Fuzi and its compound prescription could be accurate quantitative.(2) The UPLC-MS/MS method for simultaneous determination of AC, MA, HA, BAC, BMA, BHA, had a good separation degree with high response. The linear regression for Aconitum alkaloids was obtained in the range of0.1-300ng/mL. The LLOQ of the method was0. lng/mL and the LOD is0.05ng/mL. The recovery rate and matrix effect were over between85.00%-115.00%, and the analysis time was only taken7minutes. The instrument accuracy was high and the precision was good. The plasma sample was stable in24h.(3) The results showed that the fitting model for the six Aconitum alkaloids was the one-compartment model pharmacokinetics in the beagles.Conclusions:(1) The UPLC-MS/MS method we establish is fast, sensitive and accurate enough to determine the content of Fuzi, water decoction of Fuzi. It can provide a surveillance method for the Fuzi quality to improve its quality criterion.(2) The UPLC-MS/MS method to analyze Aconitum alkaloids in plasma we establish is fast, sensitive and accurate enough, and has a good linear relation, the average recovery, precision and stability is satisfactory.(3) Aconitum alkaloids are the same ingredient of Fuzi’s drug effect and toxicity. The article discusses the pharmacokinetic process of aconitum alkaloids after taken by beagle dogs and establishes UPLC-MS/MS method to analyze Aconitum alkaloids in plasma to set up Monitoring tools of Aconitum alkaloids for the use of Fuzi in clinic. The Safety guidelines of Fuzi in clinical application are illuminated by monitoring the metabolic rule of Aconitum alkaloids in vivo to figure out its mechanism of reducing toxicity and enhancing efficacy.