| Objective Rebiqing tablets tablet is a proprietary Chinese medicine for treating rheumatoid arthritis,the prescription containing Strychnos.The main effective component for treating rheumatism is strychnine and bruciine,both are toxic components.Experimental study on establishment of blood concentration of Rebiqing tablets of brucine and strychnine analysis method,Compare Strychnos to Rebiqing as a contrast,Study on rats with adjuvant arthritis?AA?model of pharmacokinetics in normal rats multiple given medicine of pharmacokinetics process,to in venstigate the pharmacokinetics of law,lay the foundation for the Rebiqing in inical safety and reasonable application of in-depth study.Methods 1.Establishment of analytical method of blood concentration of brucine and strychnine of Rebiqing.Using high sensitive ultra performance liquid chromatography tandem mass spectrometry?QExactive Orbitrap HRMS?instrument,with Brucine,strychnine and peak response value as the evaluation index,the clenbuterol hydrochloride as the internal standard,investigation of determination conditions of chromatography and mass spectrometry,the choice of mobile phase acetonitrile and 0.15%formic acid ratio and injection time,and normal rat and rat plasma containing AA specificity experiment and related methodological study.2.Study on Rebiqing tablets in brucine and strychnine in vivo rat model of AA.?1?model.were randomly selected from 36 male SD rats,in his right foot after the injection of complete Freund's adjuvant 0.1ml model was made in the paw swelling volume of rats in each group were detected the 1,3,5,8,10,12,14,15,16 day after modeling,joint swelling degree and weight,calculated swelling rate,arthritis index,weight gain rate preliminary judgment,the success of modeling.And in the pharmacokinetic experiment of blood after the blood from the abdominal aorta,determination of serum IL-1 by enzyme linked immunosorbent assay,TNF-alpha content,and take the spleen,thymus,SPSS17.0 software is applied to analyze the immune organs of spleen,thymus index,once again verify the success of modeling?2?grouping.the successful model of the rats were randomly divided into,strychnos powder,Rebiqing model group,6 rats in each group.The other 12 male SD rats were randomly divided into Rebiqing group,CompareStrychnos to Rebiqing As a contrast.?3?The amount of Rebiqing given and collection blood each group according to the amount of 0.12mg/kg Brucine,At 5,10,20,30,45,60,90,120,180,300,420,540,720min after administration,collecting orbital venous blood.Determination of brucine and strychnos vivo plasma drug treated rats by HRMS method of strychnine concentration.?4?experimental data processing method using DAS3.0 software calculation of brucine and strychnine on the pharmacokinetic parameters of drugs dynamic difference parameter calculation software SPSS17.0,analyzed by t test.3.Study of in pharmacokinetics with given medicine in normal rats of Rebiqing tablets.SD male rats were randomly divided into 12,Rebiqing group,strychnos group,6 rats in each group.According to the 0.12mg/kg of brucine dose administered with corresponding drugs,administered three times daily,orally for 6 days.From the second day,a day to second times after intragastric administration of 180,300,420min,respectively.The rats of orbital vein blood.On the morning of the seven thday after intragastric 5,10,20,30,45,60,90,120,180,300,420,540,720min,orbital blood concentration using HRMS method for the determination of brucine and soil body in plasma of rats,DAS3.0 software to calculate the strychnine medicine calculated the pharmacokinetic parameters between the pharacokinetic parameters of.SPSS17.0 the software,analyzed by t test.Results 1.Blood concentration determination method.Mass spectrometry conditions:using electrospray ion source,using sheath gas flow ion,rate:10,spray voltage?KV?:3.5,spray:0.3 current?uA?,capillary temperature?c?:320,Aux gas heater temperature:50.enbuterol hydrochloride m/z 395.199,335.177,279.161.mobile phase chromatographic conditions:30%acetonitrile:70%formic acid solution 0.15%,the column temperature was 35 degrees,0.2mL/min flow rate,sample volume 5 L,injection time 5min.of brucine and strychnine were linear in the range of 0.1215ng/mL,0.2430.75ng/mL,the average recoveries were84.93%and 83.37%.RSD is 6.06%,the matrix effects were 84.92%,84.29%,RSD 3.01%.minimum detection limit:0.06ng/mL,0.12ng/mL,strychnine,both RSD were 4.78%,4.77%.2.Study on dynamic of brucine and strychnine in AA rats.The medicine Strychnos in normal rats and AA rats were of brucine and strychnine on the pharmacokinetics of the average number of parameters were:Tmax 1.50,1.58h and 0.32,0.28h Cmax;2.26,3.45ng/mL and3.45,21,47ng/mL;AUC 35.7,29.65 and 156.56,93.82 ng·min L-1,T1/2 6.58,6.92h and2.51,2.13h,CL/F:623339?65441 L·kg-1·min-1and 810118?823325L·kg-1·min-1;Rebiqing in normal rats and AA rats nahma of brucine and strychnine pharmacokinetic parameters of the average respectively.Tmax:1.50,1.50h and 0.64,0.73h,Cmax:2.68,7.16ng/mL and5.27,21.57ng/mL;AUC0-t,16.52,52.4 and 148.42175.02 ng·min L-1,T1/2/2 4.70,5.63h and3.57,2.50h CL/F,71250?81818 L·kg-1·min-1and91349?93217L·kg-1·min-1?The group of Brucine stive The pharmacokinetics of brucine and strychnine in normal rats.AA rats conformed to the one compartment model.AA rat model between Rebiqing tablet group and Strychnos group,strychnine pharmacokinetic parameters of AUC0-t?Tmax are different.3.Normal rats were repeatedly given the Rebiqing tablet medicine.Rebiqing tablets of brucine andstrychine at steady state,from the third day after administration.The plasma concentration of strychnine was maintained at 0.140.24ng/mL,0.912.11ng/mL.Rebiqing in two group rats nahma of brucine and strychnine pharmacokinetic parameters of the average respectively.at the time of giving the pharmacokinetic parameters were mainly medicine Strychnos and Rebiqing tablets of brucine and strychnine in the Tmax were1.50,1.08h and Cmaxax respectively,4.46,18.74 ng/ml,AUC0-t-t 47.21,29.13 ng·min·L-1respectively,CL/F:88991,51352L·kg-1·min-1respectively;of brucine and strychnine in nux vomica powder in the main medicine in rats in vivo pharmacokinetic parameters Tmax were1.33,0.71h and Cmaxax respectively,3.82,15.27ng/ml,CL/F:56182201408L·kg-1·min-1respectively,and the single dose pharmacokinetic parameters were similar.Powder and Rebiqing tablets of brucine and strychnine,pharmacokinetics of medium This Agreement.Conclusion 1.Determination of the tannin contents of brucine and strychnine method in rat plasma,strong specificity,high sensitivity,accurate detection result is stable,can be used as the content of the blood concentration of brucine and scholar in preparation for the determination.2.Rebiqing brucine and brucine pharmacokinetics were consistent with one compartment model and the kinetic parameters in the AA model compared with normal rats,increase the degree of brucine and tablets in Bistrychine biological absorption speed,speed up the elimination of.Other ingredients in the compound has an effect on the process in vivo.The influence mechanism in the process,what ingredients are also need further study.3.Normal rats were repeatedly given the heat of moving Rebiqing tablet medicine.Rebiqing tablets of brucine andstrychine at steady state,from the third day after administration.The plasma concentration of strychnine was maintained at 0.140.24ng/mL,0.912.11ng/mL.at the time of giving thepharmacokinetic parameters were mainly medicine Strychnos powder and heat Biqing thiller tablets of brucine and strychnine in the Tmax were 1.50,1.08h and Cmaxax were 4.46,18.74ng/ml,AUC:47.2129.13ng·min/Lrespectively,CL/F88991,51352L·kg-1·min-1respectively;of brucine and strychnine in nux vomica powder in the main medicine in rats in viv opharmacokinetic parameters Tmax were 1.33,0.71h and Cmaxax were3.82,15.27 ng/ml respectively,CL/F:56182,201408L·kg-1·min-11 respectively,and the single dose pharmacy-okinetic parameters were similar.Strychnos and Rebiqing tablets of brucine and strychnine,pharmacokinetics of medium This Agreement. |
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