| Objective:The previous research of the research group showed that Shengxian Decoction has a good anti-heart failure effect and can reduce the cardiotoxicity of adriamycin.However,the pathology situation of Shengxian Decoction is not yet clear.Therefore,this subject studies the pharmacokinetic characteristics of the main components of Shengxian Decoction under pathological conditions,and compares the pharmacokinetics of normal rats to find metabolic differences,in order to provide a theoretical basis for clinical rational drug use.Method:1.Based on UPLC-MS/MS technology,this study established the determination method of 18 main active ingredients in Shengxian Decoction samples,which solved the quality control problem of Shengxian Decoction.Pairs include caffeic acid,vermilion isoflavones,vermilion isoflavone glycosides,ferulic acid,isovulic acid,formononetin,formononetin,mangiferin,neomangiferin,campanin saponin D,rutin,saikosaponin A,The effective components of Shengcai decoction including saikosaponin D,timosaponin A?,timosaponin B?,astragaloside IV,colicin and quercetin were measured.2.Using UPLC-Q-TOF-MS liquid-mass spectrometry technology,rats were given gavage soup extract(608 mg/kg,equivalent to 70.6g/kg crude drug)to identify the migration components in the blood.Analysis,rapid screening of drug substances.3.The use of doxorubicin to chronically accumulate heart damage,the use of multiple intraperitoneal injections,cumulative dose of 15mg/kg,to establish a rat heart failure model.4.A reliable and sensitive UPLC-MS/MS method has been established,which can determine 18 main active ingredients plasma drug concentration.5.After single administration of Shengxian Decoction to normal and model rats,collect plasma samples,determine the concentration of their main active components in plasma,understand the absorption characteristics of these components in normal and model rats,and perform pharmacokinetics Comparison of parameters.Results:1.It can be known that caffeic acid,verbascum isoflavones,verbascum isoflavone glycosides,ferulic acid,isoferuvic acid,formononetin,formononetin,mangiferin,neomangiferin,platycodin D,rutin,Bupleurum The contents of saponin A,saikosaponin D,timosaponin A?,timosaponin B?,astragaloside,colicin,and quercetin in the extract of Shengxiantang were 0.42,0.55,9.28,1.49,and 0.34,respectively,0.19,3.33,35.37,59.34,1.58,0.09,31.73,29.76,1.18,420.47,0.31,0.31,0.45 ?g/mg.From the test results,it can be seen that the higher content in the extracts is the timosa saponin B?,neomangiferin and mangiferin derived from chrysanthemum,and the saikosaponin A and saikosaponin D derived from Bupleurum chinense.2.The 18 prototype components in Shengxian Decoction extract can be identified one by one in blood components.3.Compared with the normal group of rats,the model group has poor mental performance,eating gradually decreases,and the body weight is significantly reduced;the heart's systolic blood pressure and diastolic function are significantly impaired;myocardial cells appear edema,a large number of vacuolar degeneration,accompanied by a large number Macrophage,lymphocyte infiltration and other phenomena;most mitochondrial edema,crest rupture and dissolution disappeared,mitochondrial membrane phospholipid localization showed a loss of change.4.Methodological verification results show that the standard curve shows good linearity in the concentration range to be investigated;the extraction recovery and matrix effect obtained by protein precipitation method are within the specified range;The precision and accuracy meet the analysis requirements of biological samples;the stability of the samples meets the analysis requirements of biological samples.5.The results of this experiment showed that the AUC(0-t)of the isoflavones derived from astragalus decreased and the MRT,T1/2 and Cmax of the heart failure rats after oral Shengxian Decoction were reduced compared with normal rats.AUC(0-t)of astragaloside increased,T1/2 and Cmax increased,AUC(0-t),MRT,T1/2,Cmax of formononetin all decreased,AUC of formononetin(0-t)increases,T1/2 decreases;the AUC(0-t)and AUC(0-?)of Amygdalin A?,the component derived from Anemarrhena,increases,T1/2 increases,and Cmax decreases,Amygdalin B? AUC(0-?)increased,T1/2 increased,and Cmax decreased;AUC(0-t),AUC(0-?)of rutin derived from Bupleurum chinense decreased,and MRT,T1/2,Cmax were all decreased.Decrease;AUC(0-?)of cimicifolin,a component derived from cohosh,decreases,T1/2 decreases,Cmax increases,AUC(0-?)of ferulic acid increases,T1/2 increases,Cmax decreases,The above results are statistically significant(P<0.05).Conclusion:1.Using the combination of ultra-high pressure liquid phase and high-resolution mass spectrometry,the determination of the main components in the extract of Shengxian Decoction was completed,which provided a basis for the quality control of Shengxian Decoction and ensured the quality of the prescription of Shengxian Decoction Stable and controllable.2.After oral administration of Shengxian Decoction,only the components absorbed into the blood can produce a pharmacological theory of therapeutic effect.First,the serum mass of Shengxian Decoction is studied by correlation mass spectrometry analysis of normal rat drug-containing serum and mixed standard plus serum Chemistry.3.Multiple intraperitoneal injections of doxorubicin can establish a rat heart failure model,and the rat heart function can be judged on multiple levels such as general rat behavior indicators,echocardiography,myocardial tissue pathology,and myocardial tissue ultrastructure electron microscopy.It shows that the stability of the model of adriamycin intraperitoneal injection is good and the success rate is high.4.The internal standard used in this method is amlodipine and warfarin as internal standards.The protein precipitation method is used to process plasma samples.A set of chromatographic conditions is established.One detection is performed using Agilent 1260 ultra-high performance liquid chromatography-G6460A.Triple quadrupole tandem mass spectrometer.The mass spectrometry conditions use multi-reaction detection.The simple and fast method for simultaneous detection of positive and negative ions provides a reference for later experimental detection.The 18 main components of Shengxian Decoction extract in plasma have a good linear relationship(R2>0.98),and the accuracy,precision,extraction recovery rate,matrix effect,stability,etc.are within the acceptable range,and are in line with 2015 The requirements of the "Chinese Pharmacopoeia" version 9012 biological sample analysis.Therefore,this method is suitable for the detection of plasma drug concentration of the main component of Shengxian Decoction,and can provide a reference for subsequent pharmacokinetic studies.5.The pathological state of heart failure significantly affects the pharmacokinetic behavior of Shengxian Decoction in the body.It is suggested that if Shengxian Decoction is used in reference to the normal body,the treatment will be performed according to the normal body pharmacokinetic parameters In the best case,the dosage can be adjusted according to the pharmacokinetic parameters of the body under the pathological state to ensure the therapeutic effect. |
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