Correlation Research And Evaluation Of Methods Of Drug And Safety Of Azithromycin In Mice

part one: the establishment and evaluation for the detection method of serum concentration of of azithromycin by hplcObjectiveIt establish a set of HPLC-UV analytical methods for serum in mice.Method1 The HPLC consisted of Hypersil NH2 analytical column(250×4.6mm, 5μm) as solid phase, Acetonitrile -0.01mol/L NH4Cl(Use ammonia regulation make PH=10)(90:10) as mobile phase, flow rate of 1.0ml/min,, Detected wavelength:210nm, Chromatographic column temperature:30℃;2 Basic methods of biological samples treatment: taken 100μl samples, add 50μl roxithromycin, rotated 30s,add 900μl methanol, rotated 3 min,14000r/min centrifuge 20min, Precision absorb 800μl, drying by distillation of 40℃water bath, solid residues dissolved with 100μl mobile phase, 20μl for injection.ResultsThere was a wide linear range in various biological samples (serum for 0.1 ~6mg), regression equation:Y=4.1206X-0.376(γ=0.9998), the limit of detection was 0.1μg/mL. The precision(RSD) of intra-day in three different concentrations (high, medium, low) were all≤1.45%; The precision(RSD) of inter-day in three different concentrations (high, medium, low) were all≤2.61%; The precision(RSD) of repeatability for the analysis were1.73%,1.68%,1.80%; The absolute recovery were 98.82%,100.76%,99.59%,The Plus sample recovery were 100.41%,98.42%,98.54%。ConclusionsThis experiment established serum azithromycin content determination of mice by HPLC method, the method of high sensitivity, strong specificity, sample pretreatment method is simple,rapid,it provides testing technology for biological samples and pharmacokinetic study.Part two: Correlation research of the methods of drug and the drug concentration and toxicity reactionObjectiveThe experimental study of correlation among drug dosage,Injection velocity,toxicity reaction and drug concentration, Explore the Correlation between methods of drug and the drug concentrationMethod1 Kunming mice 100, According to gender and weight are divided into 10 groups at random, Among them the nine groups ,according to the different injected dosage(442mg/kg,645mg/kg,847mg/kg), different injection velocity(10s,20s,30s), conduct final intravenous medication; The rest of the group, injected saline with the same way.2 Observe ecg and heart rate,observe the time of toxicity incubation period,the time of breathing stopped,the time of death. After the mice death, open-chest,and take blood from the heart,centrifuged and take serum. Detection the Serum concentration of of azithromycin by HPLC. Research the relationship between drug concentration and methods of drug, research the Correlation with acute toxicity reaction time.Results1 the toxic reaction after intravenous azithromycin:①The time of toxicity incubation period,the time of breathing stopped,the time of death: With the increasing dose, the reaction times is shortened, negative correlation; Along with the injection velocity down, The reaction time extended, positive correlation;②After intravenous, the survival rate of obvious change , a period of observation: With the increasing dose, Survival rate reduce;along with the injection velocity down, survival rate increase;③It occur abnormal ecg after intravenous azithromycin: Q - T interphase gradually extended,then heart attack, cardiac arrest, then mice death.2 With the increasing doses,the Blood drug concentration increases, present positive correlation; Along with the injection velocity down, Blood drug concentration lower, Present negative correlation.the comparison between each group ,(p﹤0.01).3 Blood drug concentration and the acute toxicity time is correlation, Along with the increase of blood drug concentration, the reaction time gradually shortened, Present negative correlation.Conclusions1 Intravenous at different speeds and different doses of azithromycin can be caused by abnormal ecg changes and death in mice that .It manifest that the azithromycin intravenous can cause cardiac toxicity.2 Azithromycin intravenous dose and intravenous speed all can affect blood medicine density and mice toxicity. In the meantime, give the same injection velocity, it is along with the increased doses ,the blood drug concentration rise and the various toxic reaction intensified ;Administration dosage phase at the same time, it is along with the increased injection speed, elevated blood drug concentration, various toxic reaction intensified.