Plasma Pharmacochemistry And Pharmacokinetic Study Of Daming Capsule In Rats

Background and PurposeDaming capsule is a traditional Chinese medicine for hyperlipidemia treatment.However,the bioactive components of Daming capsule are still unknown.This work aims to establish a new strategy integrating plasma pharmacochemistry,pharmacokinetics,and pattern recognition analysis,based on ultra performance liquid chromatography-electrospray ionization-quadrupole-time-of-flight-tandem mass spectrometry(UPLC-ESI-Q-TOF-MS/MS),thus facilitating modernization,internationalization,research and development(R&D)of Daming capsule.MethodsThis work first developed a high-throughput,-resolution,and –sensitivity UPLC-ESIQ-TOF-MS/MS method for analyzing Daming capsule extract and dosed plasma.In plasma pharmacochemistry study,before oral administration and 1 h after oral administration of 4 g/kg Daming capsule,1 m L blood samples were collected from the retro-orbital plexus of ten rats.Compounds absorbed into the blood from Daming capsule were screened using UPLC-ESI-Q-TOF-MS/MS combined with Marker Lynx.And all absorbed compounds were identified according to their MS and MSMS spectrum.In pharmacokinetic study,serial blood samples were collected from the tail vein before oral administration and 0.5,1,1.5,2,2.5,3,4,6,8 and 12 h after oral administration of 4 g/kg Daming capsule.Phamacokinetics of Daming capsule was operated employing UPLC-ESI-Q-TOF-MS/MS combined with semi-quantification,and six pharmacokinetic parameters were then calculated.In pattern recognition analysis,principal component analysis(PCA)and hierarchical cluster analysis(HCA)were applied to organize the compounds with the most similar pharmacokinetic properties in one cluster.By comparing the pharmacokinetic parameters between clusters,one cluster of compounds with optimal pharmacokinetic behavior were identified as potentially bioactive components.ResultsThe present study first developed an UPLC-ESI-Q-TOF-MS/MS method for identifying the absorbed compounds and monitoring the pharmacokinetics of Daming capsule.Precision,stability,and repeatability of the method were all well.In plasma pharmacochemistry study,up to 53 compounds were found to be absorbed into blood and classified into five types based on chemical structures,including 24 anthraquinones,13 phenanthraquinones,7 flavonoids,6 phenols,and 3 tetracyclic triterpenoids.The chemical structures of all absorbed compounds were successfully identified on the basis of MS and MSMS spectrum.In pharmacokinetic study,semiquantification was applied for each absorbed compounds,six pharmacokinetic parameters were then calculated,including the time for maximal concentration(Tmax),peak concentration(Cmax),area under curve(AUC0-t),elimination rate constant(Ke),biological half-life(t1/2),and total body clearance(CL).In pattern recognition analysis,dimensions of six pharmacokinetic parameters were reduced to three principle components by PCA.Absorbed compounds were then divided into clusters A,B and C based on their pharmacokinetic behaviors by HCA.To evaluate the pharmacokinetic performance of compounds in the three clusters,the parameters between these clusters were analyzed for comparison.The elimination were similar among three clusters(P > 0.05),whereas compounds in cluster B exhibited faster(small Tmax)and greater absorption(large Cmax and AUC0–t).Therefore,six anthraquinones in cluster B,namely,emodin-O-glucoside,aurantio-obtusin,aloe-emodin,rhein,emodin,and chrysophanol,were screened as potentially bioactive components of Daming capsule.ConclusionsA high-throughput,-resolution,and-sensitivity UPLC-ESI-Q-TOF-MS/MS method was first developed to quantify and identify the absorbed compounds after oral administration of DMC.A new strategy integrating plasma pharmacochemistry,pharmacokinetics,and pattern recognition analysis was successfully established for research of Daming capsule in vivo.Up to 53 absorbed compounds were identified,including 31 in original form and 22 metabolites from DMC.Among them,6 anthraquinones in original form with the most excellent pharmacokinetic performance were screened as potentially bioactive components of DMC.The present study facilitated R&D of DMC.Notably,the high-throughput and reliable strategy provided novel insights into the screening and identification of potentially bioactive components of traditional Chinese medicine.