Study On The Preparation Of Amorolfine Hydrochloride Liniment And Its Distribution In Nail Plate

Objective:The purpose of this project is to develop amorolfine hydrochloride liniment for the treatment of onychomycosis.Take commercially available amorolfine hydrochloride liniment?Product name:Loceryl^??as a control,the analytical methods,formulaiton development,transungual delivery and quality standard were studied.The results help to provide the experimental basis for the development of amorolfine hydrochloride liniment generic drugs.Methods:?1?Establishing a systematic HPLC method for determining the retention of amorolfine hydrochloride in the bovine hoof plate.The methods includes chromatographic conditions,specificity,LOD and LOQ,the calibration curves and the instrument precision.?2?Studying on the retention of amorolfine hydrochloride in bovine hoof plate when the Loceryl was applied.Choosing bovine hoof plate which was cut into slice at a thickness of1mm as a model of nail plate.A modified Franz diffusion cell was used to the transungual penetration test.After finishing the transungual penetration test,the residual on the surface of bovine hoof was washed out,and then cut the bovine hoof into pieces with scissors.After freezing with liquid nitrogen,it was smashed into powder by a Highthroughput Tissue Mill.Finally,amorolfine hydrochloride in the powder was extracted with 90%acetonitrile and determined by HPLC.The extraction recovery was determined.?3?The bioadhesive force between the liniment and the bovine hoof surface was used as an inspection index,and the dosage ranges of film-forming material?RL100?in the liniment were determined by single factor test;the drying time of the liniment was used as an inspection index,and the dosage ranges of 95%ethanol in the liniment were determined by single factor test.?4?On the basis of single factor test,performing the techological process was optimizated by taking the amount of the three main excipients in amorolfine hydrochloride liniment including RL100?A?,95%ethanol?B?and ethyl acetate?C?as the investigation factors,every factor sets 3 levels,and selecting L9?3⁴?orthogonal table to arrange the experiment.The optimum prescriptions of liniment can be determined by orthogonal design-direct analysis and ANOVA.Finally,three batches amorolfine hydrochloride liniment were prepared to verify the experiment.?5?Preliminary quality study for self-made amorolfine hydrochloride:preliminary inspection includes the content of amorolfine hydrochloride in self-made liniment,the retention of amorolfine hydrochloride in bovine hoof plate and film-forming effect.?6?The irritations of the self-made liniment on human nails and rat skin were investigated.Results:?1?Chromatographic conditions were follows:mobile phases consisted of methanol-ammonium dihydrogen phosphate solution?adjusted pH to 3.0 with phosphoric acid??70:30?.The flow rate was 1.0 ml·min^-1;The detection wavelength was 214 nm.Column temperature was 30?.Injection volume was 10?l.The LOQ and the LOD were13.0 ng·ml^-11 and 6.5 ng·ml^-1 respectively.Calibration curve for amorolfine hydrochloride was linear in the range of 5.20¹8.20?g·ml^-1?R²=0.9990?.The instrument precision is good?RSD=0.50%?.Amorolfine hydrochloride showed good specificity.?2?The average of extraction recovery was 75.25±2.46%.The average retention of amorolfine hydrochloride in the bovine hoof plate for Loceryl was 3.04±0.22%.?3?Through single factor test,the key factors and its levels were adopted:the dosage range of RL100?1.2 g¹.8 g?and the dosage range of 95%ethanol?6.5 ml⁷.5 ml?.?4?On the basis of single factor test,the optimum prescription of self-made liniment was obtained through orthogonal experiment:the film-forming material for RL100 was 1.2 g,the mixed solvent composition was 95%ethanol 6.5 ml and ethyl acetate 2.0 ml,the solubilizers for butyl acetate was 0.9 ml and the plasticizer for glyceryl triacetate was 0.1 ml.?5?The results of the preliminary quality inspection for self-made amorolfine hydrochloride showed:the average content of amorolfine hydrochloride in self-made liniment was 99.50±1.95%;the retention of amorolfine hydrochloride in bovine hoof plate was 3.07±0.08%;The film-forming effect on the bovine hoof surface between the reference liniment and the self-made liniment was completely plain,smooth and without bubble-like structure.?6?The safety of the self-made liniment was good,and have no irritation on human nails and rat skin.