Pharmacokinetics Of Two Sesquiterpene Lactones In Kudiezi Injection

In this research, Ixerin Z, 11,13α-dihydroixerin Z in Kudiezi injection were chosen as the analyzed objects and establish a method for the determination of Ixerin Z, 11,13α-dihydroixerin Z by UPLC-ESI-MS/MS. Using this method, we tested 6 batches of the 2 components in Kudiezi injection produced by 2 companies; A quantitative method was first developed for the simultaneous determination of Ixerin Z, 11,13α-dihydroixerin Z in Kudiezi injection in rat plasma, and was successfully applied to a pharmacokinetic study for comparative research of the 2 components in Kudiezi injection and monomers. The main contents and results are as follows:1. Simultaneous Determination of Two Sesquiterpene Lactones in Kudiezi injection by UPLC-ESI-MS/MSIn this chapter, simultaneous determination of Ixerin Z, 11,13α-dihydroixerin Z in Kudiezi injection by UPLC-ESI-MS/MS and the separation was performed on a Waters ACQUITY BEH C18(2.1 mm*50 mm,1.7 μm)with a gradient of water and acetonitrile in 2.0 min, multi-reaction monitoring (MRM) scanning was employed for quantification. The established analysis method was confirmed, and the results satisfied with the requirement of related regulations. The results showed that the range of Ixerin Z,11,13α-dihydroixerin Z in different batches are 4.82μg·mL-1～12.33μg·mL-1,3.07μg·mL-1～-8.82μg·mL-1, respectively.2. A analyze method was developed for the Ixerin Z, 11,13α-dihydroixerin Z in Kudiezi injection in rat plasma.In this part, a analyze method was developed for the Ixerin Z, 11,13α-dihydroixerin Z in Kudiezi injection in rat plasma by UPLC-ESI-MS/MS, internal standard is Harpagoside, and plasma samples were extracted by ethyl acetate. The separation was performed on a Waters BEH C18 column (2.1 mm* 50 mm,1.7 μm) with a gradient of water and acetonitrile in 3.0min, multi-reaction monitoring (MRM) scanning was employed for quantification in positive mode. And we make a systematic study on methodology on the analysis method, the results showed that the method is selective and sensitive and can be used to research pharmacokinetic study of Ixerin Z,11,13a-dihydroixerin Z in rats.3. Pharmacokinetic effect of Kudiezi injection on two sesquiterpene Lactones in normal rats.Rats were injected with low dose and high dose of Ixerin Z, 11,13α-dihydroixerin Z monomers, plasma samples were pretreated with ethyl acetate, study on its pharmacokinetics, and illustrate its working rules of different doses in body. The results showed that the exposure level of high dose group was higher than low dose group, which showed significant difference, the Cmax and AUC (area under the curve) value of high dose group were greater than low dose group.There was no statistical difference in t1/2 between the two groups. There were significant difference of body clearance (CLZ) and apparent volume (Vz) of distribution between them (P<0.01), and mean retention time showed no significant difference (P> 0.05).4. Comparative pharmacokinetic study on two sesquiterpene Lactones and monomers of Kudiezi injection in normal rats.Eighteen rats were randomly divided into three groups:rats in group A received Kudiezi injection, B and C received a same dose of Ixerin Z, 11,13α-dihydroixerin Z, respectively. And studying on its pharmacokinetics, the statistical analysis was performed by SAS 9.1. Explaining the effects of other ingredients during the intracorporal process. The results showed that the exposure level of Ixerin Z in group A was significant lower than monomer group, maximum blood concentration (Cmax, P<0.01) and area under the curve (AUC, P<0.001). Clearance (CLZ, P<0.001) showed significant difference, but other pharmacokinetic parameters showed no difference. Compare to monomer group, the exposure level of component in group A was increased (Cmax, P<0.05), clearance decreased (CL2, P<0.001), mean residence time decreased (MRT, P<0.05).5. Determination of bovine serum albumin binding rate on two sesquiterpene LactonesThe equilibrium dialysis combined with HPLC to determine the bovine serum protein binding rate of Ixerin Z,11,13α-dihydroixerin Z, the result showed that mass concentration of Ixerin Z, 11,13α-dihydroixerin Z is 0.032,0.128,0.256 mg·mL-1,0.038,0.152,0.303 mg·mL-1, respectively, their protein-binding rate is 51.21%,54.72%,52.14% and 50.63%, 52.82%,50.28%, respectively. The two sesquiterpene Lactones have moderate intensity protein-binding rate.