A new method for measuring gentamicin in liposomes fluorometrically is described. The assay is based on the reaction between the amino groups in the gentamicin molecule and o-phthaldialdehyde (OPA), under basic pH conditions; the product's fluorescence can be read directly on a simple fluorimeter. The effects of several factors (time of reaction, volume of the OPA reagent, and product stability) were investigated. The standard curve was linear in the concentration range of 0.5-4.0microg, showing an excellent determination coefficient of r(2)=0.99. Additionally, the influence of different liposomal lipids on gentamicin determination was tested. Liposomal lipids containing no free amino groups (PC, Chol, DOTAP) have no influence on the reaction when present in the reaction mixture. In contrast, amino groups containing lipid (SA) showed intense method interference. Therefore, a method of lipid extraction was adapted to remove undesired lipids. The described method was successfully utilised during 2 years of liposomal gentamicin experiments.

译文

描述了一种用荧光法测定脂质体中庆大霉素的新方法。该测定法基于在碱性pH条件下庆大霉素分子中的氨基与邻苯二甲醛 (OPA) 之间的反应; 可以在简单的荧光计上直接读取产品的荧光。研究了几个因素 (反应时间,OPA试剂的体积和产物稳定性) 的影响。标准曲线在0.5-4.0microg的浓度范围内呈线性,显示出极好的r(2)= 0.99的测定系数。此外,还测试了不同脂质体脂质对庆大霉素测定的影响。当存在于反应混合物中时,不含游离氨基的脂质体脂质 (PC,Chol,DOTAP) 对反应没有影响。相反,含脂质 (SA) 的氨基表现出强烈的方法干扰。因此,采用了一种脂质提取方法来去除不需要的脂质。所描述的方法已在2年的庆大霉素脂质体实验中成功使用。

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