| Objective:(1)To investigate the anti-depression effects of ifenprodil tartrate on the forced swimming mice model.(2)To establish the quantitative analysis method of ifenprodil tartrate in biological sample(SD rat plasma).(3)To investigate the steady state plasma concentration of ifenprodil tartrate after intraperitoneal injection in five half-times and its Pharmacokinetic characteristics.Methods:Forced swimming test(FST)on mice was used to investigate the anti-depression effect of ifenprodil.(1)Test of the onset time and the duration time of ifenprodil tartrate in the antidepressant effect:one hundred and fifty mouse were randomly divided into fifteen groups(n= 10),including the experimental groups(1 mg·kg-1 ifenprodil tartrate),the positive control groups(2 mg·kg-1 ketamine)and the control groups(NS).The forced swimming test were taken after 30 minutes,60 minutes,120 minutes,240 minutes and 300 minutes respectively and the immobility time of mouse were recorded.(2)Test of the dose-effect relationship of ifenprodil tartrate in the antidepressant effect:One hundred mouse were divided randomly into ten groups(n=10),including the experimental groups(0.2 mg· kg-1,1 mg· kg-1 and 5 mg·kg-1 ifenprodil tartrate),the positive control groups(2 mg·kg-1 ketamine)and the control groups(NS).The forced swimming test was taken after 120 minutes and 240 minutes respectively.(3)Test of the antidepressant effect evaluation of ifenprodil tartrate on repeated dose:sixty mouse were randomly divided into six groups(n=10),including the experimental groups(5 mg-kg-1 ifenprodil tartrate)and the control groups(NS).The experimental group mouse were intraperitoneal injectioned ifenprodil tartrate for two days,four days and six days,respectively.The forced swimming tests were taken after 240 minutes at the last day of injection.LC-MS/MS method was used to detemine the ifenprodil tartrate in biological sample(SD rat plasma).The specificity,accuracy,precision,inearity range and freeze-thaw stability of LC-MS/MS method will be investigated.Biological sample(SD rat,SPF,half male and female,180-200g).The rats were intraperitoneal injectioned ifenprodil tartrate for five times with the half-time of 2.42h.After the fifth dose,take the plasma samples in lOmin,25min,45min,75min,90min,1 OOmin,120min,180min,240min,360min.The plasma concentration of ifenprodil in rat will be determined with the LC-MS/MS method.Analyse the absorption and disposition of ifenprodil in rat by the use of Drug and Statisties software 2.1.Result:(1)The onset time and the duration time of the effect of anti-depression of ifenprodil were respectively 120 minutes and 240 minutes.There was a positive correlation between the dose-dependent relationship and the antidepressant effect of ifenprodil tartrate in the range of 1 mg-kg-1?5 mg kg-1.The duration time on repeated dose still was 120 minutes.(2)LC-MS/MS conditions:C18 column(Waters,1.7?m,2.1×50 mm),mobile phase:A(methyl alcohol),B(5mmol ammonium acetate),gradient elute,flow rate:0.3 mL/min,injection volume:2 ?L,parsing time:6min.To use the ESI ion source,positive scan patterns,the best detection ions of ifenprodil and KCZ are 326.2?531.0.The retention times of ifenprodil and KCZ are 3.10min?3.70min.The method had a good linear relationship from 5 to 100 ng/mL.The lower limit of quantitation is 5ng/mL.The intra-and inter-day RSD are less than 15%and the biological sample is steady after three times freeze-thaw.Conclusion:(1)The effect of anti-depression on the forced swimming mice model for ifenprodil tartrate was accurate and the duration time was stable enough on repeated dose.(2)Analyse the plasma concentration of ifenprodil in rat will be faster and exacter with the use of LC-MS/MS.(3)The half-life of ifenprodil is 2.86 h. |
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