| Objective1.To explore the best preparation method of the test sample of Wusheng decoction;Explore The influence of the content of aconitine that Packet Fried?centrifugal process on During the process of the decoction;Survey the methodology the three kinds of type monoester aconitum alkaloids in Wusheng decoction;Observing the change of the content of total aconitum alkaloids in the process of decoction;Observing the stability of Wusheng decoction under the condition of low temperature and Corrosion proction?2.Observing acute toxicity recation of the test sample of Wusheng decoction,understanding the acute toxic effect dose?Through subacute toxicity test in 30 days,observing the toxic reaction of rats and its dose response relationship;Whther creating the drug discontinuation delay toxic effects after stopping the drug?Methods1.Make content determination of test sample of alcohol sedimentation technique?alcohol sedimentation technique and 0.01%HCL?evaporation of dryness?extraction of ether by HPLC,Observing the peaks and trailing factor;Use the technology of cloth Fried and centrifugal,Observing the peak of seven test samples at the time points of 30min?60min?90min?120min?150min;Determining the content of three kinds of monoester aconitum alkaloids by HPLC,establish a methodology to investigate;Observing the content changes of aconitum alkaloid at the point of 0h?0.5h?1h?1.5h;Determining the content of six monoester aconite alkaloid within six months?2.In acute toxicity test,Using the method of maximum dosage,oral Wusheng decoction singlly,according to the 30mL/kg body weight lavage three times in a row in 24 hours,interval at least 4 hour,observing the appearance?behavior?mental?secretion and viscera anatomic pathological changes of SD rats during the dosing period and after 14days?In subacute toxicity test,128 SD rats were devided into blank?low?medium and high does of four groups,each group has half group of male and half group of female,each group of sex has 4 cage?Repated oral doses of Wusheng decoction at3.4?low??6.8?middle??13.5?high?g.kg-1 in SD rats were adminstered for 30 days,withdrawal recover 28 days,the end of 29 days and convalescence,for an aphthalmologist and urine tests of SD rats,the end of 30 days and recovery period ends,for part of hematology,blood biochemistry and pathological examination in SD rats.Results1.Compared other three kinds of reference substance,the test sample using the method of ether extraction,hold a more stable peak area at the point of 20min?24min?26min;Package fried?centrifugal process had no effect on the alkaloid constituents of Wusheng decoction;Benzoyl new aconitine?Benzoyl monkshood alkali?Benzoyl monkshood alkali can be selected as the indicators of content determination of Wusheng decoction;The content of three kinds of monoester aconitum alkaloids experienced down slightly first and then rise gradually?It cant be detected completely at the point of 1 hour;The content of three kinds of monoester aconite alkaloids of Wusheng decoction was stable in the past six month?2.In acute toxicity test,there was no death in rats,the rats accpetted the test sample was not seen obvious abnormity,including the appearance?behavior?spirit?response to simuli?secretion.The largest crude drug dose of Wusheng decoction was 40 5g/kg?In the subacute toxicity text,each dose group of SD rats' s general form was good,its weight steadily rising,the food intake,ophthalmology was not apparently unusual;Compared with the solvent control group,SG values in each dose group during 29 days,high dose group of male mice was statistically significant?P<0.05?;End of recovery period,the difference between solvent control group of high dose group of females was statistically significant?P<0.05?;In 29 days,compared with the solvent control group,the urinary pH value of low?medium group of female was statistically significant?P<0.05?;In 30 days,The HGB and HCT of low dose groups of female mice was lower,compared with the solvent control group,its difference was statistically significant?P<0.05?;End of recovery period,compared with the solvent control group,The difference of the MCHC of high dose group was statistically significant?P<0.05?;At the end of recovery period,The PT' s difference between the solvent control group and the middle dose groups of female was statistically significant?P<0.05?;In 30 days,the ALT of high dose groups of female mice was higher,compared with solvent control group,Its difference was statistically significant?P<0.05?;At he end of recovery,the GLU of low dose groups of female was lower,compared with solvent control group,its difference was statistically significant?P<0.05?;In 30 days,the Urea of low dose groups of female mice was higher,compared with solvent control group,its difference was statistically significant?P<0.05?;At the end of recover period,the TG of high dose group of male mice was lower,compared with solvent control group,its difference was statistically significant?P<0.05?;At the end of recover period,the K+ of high dose group of male mice was higher,compared with solvent control group,its difference was statistically significant?P<0.05?;The spleen coefficient of high dose groups of female mice was higher,compared with solvent control group,its difference was statistically significant?P<0.05?;The rest dection of blood and urine also the major viscera pathological anatomy wasn't obvious abnormal?Conclusion1.Preliminarily choose the method of diethyl ether in low temperature for many times to make the test sample;Determined the method of sheet and fried;The method of HPLC was reliable and had a good accuracy;Preliminary survey the stability of Wusheng decoction was well under the condition of low temperature and corrosion protection.2.Under this experimental condition,give Wusheng decoction(40.5g.crude drugs.kg-1,for clinical decodes measuring 96.4 time)three times within 24 hours,didn't seen related toxicity reaction?All individual data were on the scope of the background and its dependence were on dose,The test Wusheng decoction lavage on SD rats for 30 days didn't see obvious toxic effects?The no observed adverse effect level?NOAEL?of Wusheng decoction was 13.5 crude drugs·kg-1.d-1. |
PDF Full Text Request