A Study On The Intraocular Toxicity And Side Effects And Pharmacokinetics Of Eicosapentaenoic Acid After Intravitreal Injection In Rabbits

Part one The effect of visual evoked potential and electroretinogram of eicosapentaenoic acid after intravitreal injection in rabbitsObjective:To study the effect of visual evoked potential and electroretinogram on rabbits’ eyes received intravitreal injection of different doses of EPA.Methods:Twenty-four rabbits were randomly averagely divided into 4groups.The experimental eyes received intravitreal injections of EPA at different concentrations(0.01mg/0.1ml,0.1 mg/0.1ml,1.0mg/0.1ml)and 0.1%DMSO in 0.1ml.The control eyes received 0.1ml intravitreal injections of0.9% saline solution.After injection,P-VEP、ERG were performed on 1 d,3 d,7 d and 14 d.Result:In all experimental groups,the VEP ’s latencies of P100 and the ERG’s b-wave amplitudes didn’t have significant difference compared to the control groups.Part two The intraocular pharmacokinetics of eicosapentaenoic acid after intravitreal injection in rabbitsObjective:To study the pharmacokinetics of eicosapentaenoic acid on rabbits’ eyes after intravitreal injection.Methods:Thirty rabbits were randomly divided into standard and quality control group(3 rabbits)and experimental groups(27 rabbits).The experimental groups were randomly averagely divided into 9 groups.The standard and quality control groups were to make the vitreous samples of blank and quality control.0.1ml(0.01mg)EPA was injected into the vitrous of the experimental rabbits and didn’t administrated in the standard and quality control group.The both eyes from experimental rabbits were enucleated at 5、30、90、150、240、390、510、690、720min after injection of EPA respectively to make the vitreous samples.The concentration of EPA in the vitrous samples were detected by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS).The main parameters of pharmacokinetics were calculated by DAS pharmacokinetics software.Results:The peaks of EPA and gliquidone were well.Blank vitreous didn’t interfere with the determination of endogenous substances.The peak of EPA was appeared at 1.94 min.The peak of gliquidone was appeared at0.75 min.The concentration-time data of EPA in rabbit vitreous was subject to two-compartment model that had a lag time.The calibration curve of EPA in presented linear through 0.0200μg/g to 5.00 μg/g with a regression formation as follow:Y=0.405×C+0.00152(r=0.9972,n=8).The correlation index was between 0.9972 to 0.9981.The recover rate of the quality control samples of low,mean and high concentration respectively was(105.6±6.9)% 、(102.3±2.3)% and(94.6±2.1)%.The recover rate of the internal starderd(gliquidone)was(91.6±3.6)%.The relative standard deviation(RSD)of intraday of the lower limit of quantification(LLOQ)and the quality control samples of low,mean and high concentration respectively was 10.7%、7.0%、2.1% and 2.2%.The RSD of intre-day respectively was 14.0%、11.6%、5.2%and 4.1%.The half-time of EPA in vitreous was 4.72 h.Conclusion:1 It’s safe to inject EPA into the rabbit vitreous when the concentration is less than 1.0mg/0.1ml.2 The method of HPLC-MS/MS to detect the concentration of EPA had a better specificity.The concentration-time data of EPA in rabbit vitreous was subject to two-compartment model that had a lag time.The half-time of EPA in vitreous was 4.72 h.