| Objective:1. To establish an HPLC method for the determination of propotol in human plasma and to provide basis for population pharmacokinetics study in the second chapter.2. To study population pharmacokinetics of propofol in Xinjiang uygur autonomous region and to estimate the pharmacokinetic parameters, analysis factors such as height, weight, ange, ethnic influence on the pharmacokinetic parameters.3. Explore the feasibility of using mocrodialysis technology for propofol sampling, explore the influence of the infusion speed, the composition of perfusion fluid, the medium of microdialysis on the probe extract efficiency, to provide a scientific basis for the using of microdialysis for lipopohilic compounds. Methods:1. HPLC experiment:with carbamazepine as the internal standard, the separation was performed with a Hanbon Science&. Technology C18 (4.6 mmx250 mm,5μm) column using a binary gradient elution of a mixed solution of acetonitrile and methanol (acetonitrile:methanol=80:20, v/v)-1% trifluoroacetic acid water (ph=4.0),0～7 min (50:50, v/v),7～9 min (50:50～70:30, v/v),9～22min (70:30, v/v), flow velocity was 1.0 m 1/min and the wavelength of detector was set at 274 nm, injected sample volume was 20μL, and the column temperature was set at 40 ℃.2. Population pharmacokinetic experiment:totally 95 patients were collected from anesthesiology, third affiliated hospital of Xinjiang medical university (Xinjiang Uygur autonomous region cancer hospital). The patients were divided into three groups according to their nations. The Han group had 54 patients, the Uygur group had 32 patients, the Kazak group had 9 patients. After blood samples were pretreated, propofol concentrations of plasma were determined by high performance liquid chromatography with ultraviolet detection. Using Excel and SPSS 20.0 for data mining and data statistics, adopting Monolix 4.3.3 for population pharmacokinetics analysis. On the basis of selection of three compartment model, some parameters, such as age, gender, weight, national fixed effects corresponding to the central chamber clearance (CL), the central chamber apparent volume of distribution (V1), light surrounding ventricular clearance (Q2), light surrounding apparent volume of distribution (V2), deep surrounding ventricuar clearance (Q3) and deep surrounding apparent volume of distribution (V3) these six pharmacokinetic parameter mapped analysis respectively. If there was a certain correlation, then introduce them into the model one by one. The forward inclusion and the backward elimination were adopted to establish the regression model. The stochastic approximation expectation-maximization (SAEM) algorithm was used to set up the final model. The intuitive prediction test (VPC) was used to internal validate final model.3. Microdialysis experiment:recovery experiment and delivery experiment were uesd to evaluate probe’s extract efficiency, high performance liquid chromatography (HPLC) method was used to detect analytes concentration. Investigating the influence of perfusion fluid in different infusion speed (0.5,1.0,1.5,1.0,1.5μl/min) for propofol emulsion (deprivan) probe extract efficiency and the feasibility of microdialysis sampling of propofol; examining under the same experimental conditions, the effect of different microdialysis medium (deprivan and propofol standard solutions) on proble extract efficiency; discussing the impact of perfusion liquid which contained different concentrations of hydroxypropyl-beta-cyclodextrin on deprivan probe extract efficiency and the impact of of perfusion liquid which contained different concentrations of hydroxypropyl-beta-cyclodextrin (1%,10%,20%, w/v) on propofol standard solutions probe extract efficiency. Results:1. HPLC experiment:The linear range of cefoselis was 0.10～25 (r=0.9998) with the lower limit of quantization of 0.10μg/mL.The methodological recovery of three levels of concentration of high (12.50μg/mL), medium (1.56μg/mL) and low (0.19μg/mL) were between 85% and 115%, the average recovery rate RSD were less than 10%.2. Population pharmacokinetic experiment:the six population pharmacokinetic parameters were as follows:CL= 7.78-0.069* AGEi, unit:L/min; V1=13.4 L; Q2=1.47+0.749*log (BWi/63), unit: L/min; V2=54.9 L; Q3=0.1 L/min; V3=171 L. Weight and Q2 has obviously positive correlation, and V1, V2 has certain correlation; age and CL has obvious negative correlation, and Q2, V1 has certain relevance.3. Microdialysis experiment: ①the influence of the infusion speed:deprivan under various flow velocity, the recovery values of the recovery experiment and delivery experiment have significant difference (P< 0.01). At the speed of 1.0 μl/min, the mean values of extract efficiency of recovery experiment and delivery experiment were higher than other flow velocity, the mean value of recovery experiment was 0.33%, the mean value of delivery experiment was 45.68%, and the correlation of the two experiments was the strongest (R2=0.96).② when perfusion fluid had no hvdroxvnronvl-beta-cvciodextrin. comnared the extract efficiencv values of denrivan with propofol standard solutions:the values of delivery experiment were higher than the values of recovery experiment of deprivan; the values of delivery experiment were higher than the values of recovery experiment of propofol standard solutions; the values of deprivan were higher than the values of propofol standard solutions of recovery experiment; the values of propofol standard solutions were higher than the values of deprivan of delivery experiment.③ when perfusion fluid had hydroxypropyl-beta cyclodextrin, compared the extract efficiency values of deprivan with propofol standard solutions:for deprivan, the values of delivery experiment were higher than the values of recovery experiment in the six concentrations; for propofol standard solutions, the values of delivery experiment were higher than the values of recovery experiment when concenrtation was 1%, (w/v), the values of recovery experiment were roughly higher than the values of delivery experiment when concenrtation was 10%, (w/v), the values of delivery experiment were roughly higher than the values of recovery experiment when concenrtation was 20%, (w/v).④ when the concentration of hydroxypropyl-beta cyclodextrin were same (1%,10%,20%, w/v) for deprivan and propofol standard solutions, the comparison of deprivan and propofol standard solutions extract efficiency: for recovery expriment, the values of propofol standard solutions were roughly higher than the values of deprivan; for delivery expriment, the values of propofol standard solutions were roughly higher than the values of deprivan when concenrtation was 1%, the values of deprivan were higher than the values of propofol standard solutions when concenrtation was 10%, when concenrtation was 20%, the values of deprivan were higher than the values of propofol standard solutions during 0.5-3.5 h and the values were similar during 4.0～11.0 h.⑤ the effect of adding hydroxypropyl-beta-cyclodextrin on deprivan and propofol standard solutions:for recovery experiment:the values of extract efficiency increased with the increase of hydroxypropyl-beta-cyclodextrin’s concentration in 0.5%～10%(w/v), but decreased with the increase of hydroxypropyl-beta-cyclodextrin’s concentration in 10%～20%(w/v) for deprivan; propofol standard solutions had the same trend. For delivery experiment:the influence of hydroxypropyl-beta-cyclodextrin’s concentration to extract efficiency was little in 0.5%～10%(w/v), but the values of extract efficiency reduced slightly when the concentration were 15% and 20%(w/v) for deprivan. The influence of hydroxypropyl-beta-cyclodextrin’s concentration to extract efficiency was little in 1%(w/v), but the values of extract efficiency reduced significantly when the concentration were 10% and 20%(w/v) for propofol standard solutions. Conclusion: 1.HPLC experiment:the method provides a sensitive, accurate, precise and reliable analytical procedure for the therapeutic drug monitoring of propofol in clinic and population phamacokinetic studies.2. Population pharmacokinetic experiment:by investigating the influence of demographic characteristics to xinjiang tumor patients’ population pharmacokinetic parameters of propofol, and using Monolix software established population pharmacokinetic model of propofol in conformity with the characteristics of the xinjiang region tumor patient. The population pharmacokinetic parameters which estimated by the this research could well reflect characteristics of xinjiang tumor patients.3. Microdialysis experiment:microdialysis extract efficiency of lipotropy substances propofol is lower than the average recoveries of hydrophily substances. The results show that the values of recovery experiment and delivery experiment exist significant differences and correlations. So the delivery experiment should be used in place of the recovery experiment for probe calibration. Add hydroxypropyl-beta-cyclodextrin in perfusion fluid can improve the extract efficiency of recovery experiment. |
PDF Full Text Request