Quality Control And Poly-pharmacokinetic Study Of Peiyuan Tongnao Capsule

Peiyuan Tongnao capsule?PTC?is composed of Polygoni Multiflori Radix Praeparata,Rehmanniae Radix Praeparata,Asparagi Radix,vinegar Testudinis Carapax et Plastrum,etc.It is widely used for the treatment of ischemic stroke,kidney deficiency,and blood stasis in China.The components of PTC are extremely complex,mainly including organic acids,flavonoids,stilbenes,phenylethanols,iridoids,monoterpenoids,nucleosides,and anthraquinones.However,current research on the overall quality control and poly-pharmacokinetic of PTC is very limited.Therefore,it is urgent to establish an efficient,sensitive and reliable analytical modern method to achieve the overall quality control and poly-pharmacokinetic of PTC.Objectives:1.An offline two-dimensional liquid chromatography/quadrupole-Orbitrap mass spectrometry?2D-LC/Q-Orbitrap-MS?approach was established for multicomponent characterization of PTC.2.Ten quality control indicators?citric acid,rehmannioside D,echinacoside,paeoniflorin,verbascoside,liquiritin,2,3,5,4'-tetrahydroxy stilbene-2-O-?-D-glucoside,cinnamic aldehyde,glycyrrhizic acid,and emodin?were choosed for the quality control of PTC;An LC-MS/MS quantitative analysis method was established based on the quality control indicators.3.The identification of blood components in plasma and the poly-pharmacokinetic study of ten components after administrated of PTC in rats were carried out.Methods:1.An offline 2D-LC system configuring an Xbridge Amide column and an HSS T₃column was constructed by optimization of stationary phase,mobile phase,column temperature,and elution gradient conditions.Eleven fractions of PTC,after the first-dimensional chromatography,were further analyzed by UHPLC/Q-Orbitrap-MS using a rapid negative/positive switching mode.The multicomponent characterization of PTC were realized by means of high resolution MS data quasi-element synthesis,standards,literatures and online databases,etc.2.The fingerprint analysis of seven constituted herbal medicines of PTN was used to construct ten quality control indicators of PTC.A UHPLC/PRM^admethod was established to quantify the ten compounds in fourteen batches of PTC.3.The detection mode of UHPLC/PRM was choosed for the determination of compounds.The biological samples were tested and validated in methodology.Rats were divided into three groups?n=6 for each group?.High does group?H?,middle dose group?M?,low does group?L?were orally administered with PTC?2.84g/kg?5.67 g/kg?11.34 g/kg?,respectively.Blood samples were collected from ophthalmic venous plexus after dosing.The plasma contents of the ten compounds were quantitatively analyzed by UHPLC/PRM.For the poly-PK studies,the drug concentration-time curves were plotted and the pharmacokinetic parameters were calculated.Results:1.The orthogonality of the offline 2D-LC system was 0.63,and the peak capacity was 6930.A total of 178 compounds were isolated and identified,of which96 compounds were identified.2.Based on the fingerprint analysis of the seven constituent herbs of PTC,ten quality control indicators were established.Then,a quantitative method for ten compounds was established and the methodology was verified.This method was applied to quantify ten quality control indicators in multiple batches of PTC samples.The results showed that the content of citric acid was the highest,but the content of different batches fluctuated widely,ranging from 47.899-89.022?g/10 mg.While the content of other compounds fluctuated within a relatively small range,the contents of rehmannioside D,echinacoside,paeoniflorin,verbascoside,liquiritin,2,3,5,4'-tetrahydroxy stilbene-2-O-?-D-glucoside,cinnamic aldehyde,glycyrrhizic acid,and emodin were in the range of 1.785-2.470?g/10 mg,29.497-34.824?g/10 mg,3.338-7.158?g/10 mg,4.435-7.376?g/10 mg,1.012-1.315?g/10 mg,19.083-26.195?g/10 mg,11.175-18.991?g/10 mg,1.544-2.247?g/10 mg,and 0.526-0.740?g/10mg.3.A UHPLC/PRM-MS method was established,and the methodology met the quantitative requirements of FDA for biological samples.Then,the method was used for the detection of plasma samples at different time points after oral administration of the contents of PTC in rats.The results showed that t_1/2of the low,middle,and high dose groups of the ten compounds after oral administration was independent of the dose,and the concentration peaks were observed at 0.092-1.017 h after administration.The content of each component in the body was quite different,and the AUC of aucubin and salidroside were significantly higher than other components.The MRT of the ten compounds was above 1.5 h,indicating that the elimination rate was slow.Conclusions:In order to achieve the overall quality control of Traditional Chinese Medicine preparations,PTC was used as an example to establish three methods for its qualitative,quantitative analysis,and poly-pharmacokinetic study.First,a reliable and effective analytical method based on offline 2D-LC/Q-Orbitrap-MS was developed for the rapid profiling and identification of compounds from PTC.Due to its good orthogonality and peak capacity,178 compounds were initially detected.Subsequently,by comparison the fingerprint analyses of PTC and its formula composition under the same condition,ten compounds were finally selected as the quality control indicators.Then,based on the PRM^admode in UHPLC/Q-Orbitrap-MS,10 quality control indicators in 14 batches of PTC samples were quantified simultaneously.Finally,20 compounds were absorbed through gastrointestine in rats after administration PTC,and poly-pharmacokinetic of ten components based on the LC-MS method were further developed.In summary,the comprehensive studies of PTC provided a methodological reference for its quality control and provides a certain scientific basis for further pharmacological and clinical PK research.