The Study On Pharmacokinetics Of Yao Tong Ning Capsule After Intragastric Administration In Rabbitts

Yao Tong Ning capsule has the significant efficacy on treating lumbar disc herniation, sciatica, lumbar muscle strain, lumbar muscle fiber inflammation, arthralgia by rheumatism, induced by cold-damp stasis blocking meridians. The nux vomica in this compound is the key rheumatic drug, and has strong pharmacological activities as well as great toxicity. The strychnine is the main toxic element of nux vomica. So we carried out the research on the pharmacokinetic of rabbits by gavage Yao Tong Ning capsule indexed by strychnine, in order to exploring the variation of strychnine in vivo and the influence with yellow wine. These can reveal the action characters of Yao Tong Ning capsule, and guide reasonable clinical prescription. The first part The study on analysis method of strychnine in the rabbit plasmaObjectiveTo establish the method to determine the strychnine in the rabbit plasma.MethodDetermining the chromatographic conditions of strychnine in the rabbit plasma with the High Performance Liquid Chromatography( HPLC) by investigating the detection wavelength, kinds and p H of the mobile phase, and differents of columns; then determining the treatments of plasma sample by investigating the alkali liquor, extractant, time of vortex and ultrasonic, and breaking film indexed by extraction efficiency of strychnine; moreover carriying out the method validation.ResultThe strychnine in the rabbit plasma was achieved by HPLC with the column of Agilent 5 TC- C18( 250 mm ′ 4.6 mm, 5 μm) and Agilent TC-C18( 12.5 mm ′ 4.6 mm, 5 μm) guard column at 25℃; Mobile phase consists of methanol- 0.01 mol/L sodium heptane sulfonate and 0.02 mol/L potassium dihydrogen phosphate( adjusted p H to 3.0 with 10% phosphoric acid)- acetonitrile( 0.5 : 79.5 : 20.0); detection wavelength was 254 nm, and the flow rate was 1.0 m L/min, the injection volume was 70 μL.The treatments of the plasma sample : Thaw the plasma sample at 37℃ water bath. And absorb the sample 1.5 m L precisely to 50 m L centrifuge tube, then add acetonitrile and 2 mol/L sodium hydroxide solution 0.5 m L separately, vortex 30 s, and ultrasonic 15 min in cold water, then add dichlormethane 5 m L, vortex 1 min, centrifuge 10 min in speed 5000 r/min, and collect the extract liquid; add dichlormethane 4 m L to the raffinate again for extraction repeatly; combine the extract liquid and blow dry it at 37℃ water bath by nitrogen, and re-dissolve it by 100 μL mobile phase, vortex 1 min, then centrifuge 15 min in speed 12000 r/min, finally take the supernatant.Strychnine reached its peat at 20 min, and the plasma endogenous materials had noninterference with determination; moreover there was no residual effect when injecting continuously. The strychnine was in good linearity within 0.60~200.41 μg/L, and all of the r > 0.99. The accuracy and precision of within run and between run were well. The standard reserving solution had good stability at room temperature for 24 h and-20℃ for 30 d; the standard solution also had good stability at 4℃ for 30 d; and the quality-control samples had excellent stability at room temperature for 24 h, freeze-thaw circle as well as-20℃ for 10 d; the solution of quality-control samples had good stability at room temperature for 24 h.ConclusionThe detection method and treatments of strychnine in plasma are simple, the results of method validation meet the requirement in quantitative analysis of biological; and they can be applied in detection for strychnine in rabbit plasma. The second part The study on pharmacokinetics of Yao Tong Ning capsule after intragastric administration in rabbittsObjectiveTo detect the pharmacokinetic parameters on strychnine of semen strychni powder( modulation) capsule, Yao Tong Ning capsule, Yao Tong Ning capsule with yellow wine in rabbitts; and compare the group diference.MethodProcessing data by DAS 3.2.7 based on the blood analysis, and detecting the parameters of compartment model and non-compartment model, then making t test.ResultThe compartment model parameters of strychnine after gavage Yao Tong Ning capsule : AUC(0-t) was 319.47±59.01 μg/L·h, AUC(0-∞) was 345.89±86.69 μg/L·h, Cmax was 42.73±13.63 μg/L, Tmax was 2.64±1.23 h,Ka was 1.61±1.56 per hour, t1/2γ was 3.34±0.90 h, V1 was 893.82±742.91 L/kg. The statistical moment parameters of strychnine after gavage Yao Tong Ning capsule : AUC(0-t) was 294.62±73.79 μg/L·h, AUC(0-∞) was 319.90±98.90 μg/L·h, Cmax was 49.12±11.16 μg/L, Tmax was 2.70±0.91 h, MRT(0-∞) was 8.41±3.82 h, t1/2z was 5.60±3.70 h, Vz/F was 9562.05±4188.78 L/kg, CLz/F was 1426.15±526.28 L/h/kg.The analysis results of compartment model parameters : The pharmacokinetic behaviors of strychnine in three groups were all three-compartment open model after gavage semen strychni powder( modulation) capsule, Yao Tong Ning capsule and Yao Tong Ning capsule with yellow wine; there was no significant difference in absorption rate constant and elimination half-life between three groups. Distribution of strychnine in Yao Tong Ning capsule group was close to Yao Tong Ning capsule with yellow wine group while there was big difference to semen strychni powder( modulation) capsule group according to the apparent volume of distribution. The AUC as well as Cmax of Yao Tong Ning capsule group decreased compared with semen strychni powder( modulation) capsule group, and there was statistical difference between them.The analysis results of statistical moment parameters : The AUC as well as Cmax of Yao Tong Ning capsule group decreased compared with semen strychni powder( modulation) capsule group, and there was statistical difference between them, which was same to the analysis results of compartment model. The apparent distribution volume as well as total clearance of strychnine in Yao Tong Ning capsule group and Yao Tong Ning capsule with yellow wine group increased significantly compared with semen strychni powder( modulation) capsule group. And there was no significant difference between three groups on Tmax as well as t1/2z.ConclusionThe pharmacokinetic behaviors of strychnine was three-compartment open model after gavage Yao Tong Ning capsule, and the Tmax was about 2.7 h. The other drug flavors in Yao Tong Ning capsule may have significant effects on absorption and distribution of strychnine after compatibling with semen strychni. And The distribution of stychnine changed further after compatibling with yellow wine.