柱前衍生化反相高效液相色谱法测定双硫仑角膜渗透率

A RP-HPLC Method with Pre-column Derivatization to Determine Disulfiram Cornea Transpenetration

目的 :建立一种柱前衍生化高效液相色谱新方法 ,通过测定人工房水中二乙基二硫代氨基甲酸的浓度 ,计算出双硫仑角膜渗透率。方法 :由于样品中二乙基二硫代氨基甲酸不稳定 ,且难以直接测定 ,故以对溴苯甲酰甲基溴进行衍生化并检测其衍生化产物 ,选用反相高效液相色谱 -紫外检测法定量。结果 :该方法的线性范围为 1 0～ 40 0 μg·mL-1,加样回收率为 97 44 %～ 98 75 % ,日内精密度为 0 10 %～ 0 81%。结论 :方法准确 ,操作简单 ,重现性好 。

Objective:A new RP-HPLC method with pre-column derivatization has been developed to determine diethyldithiocarbamic acid(DDC) in artificial aqueous humor for calculating disulfiram cornea transpenetration.Method:Because DDC in the sample was unstable and difficult to determine directly, p -bromophenacyl bromide( p -BPB)was used to react with DDC and the derivatization product was determined by RP-HPLC with ultraviolet detector.Results:The linear range of the assay was from 1 0 μg·mL -1 to 40 0 μg·mL -1 .Recovery of method was 97\^44%～98\^75% and the intra-day precision of method was 0\^10%～0\^81%.Conclusion:The method was proved to the accurate,convenient and reproducible.It is suitable for determination of biological samples.