Faculty of Pharmacy, Department of Analytical Chemistry, Bezmialem Vakif University, Fatih, Istanbul, Turkey.
Luminescence. 2014 Feb;29(1):87-91. doi: 10.1002/bio.2507. Epub 2013 Mar 21.
A simple, sensitive and selective spectrofluorimetric method has been developed for the determination of tobramycin (TOB) in human serum and pharmaceutical preparations. The method is based on the reaction between the primary amino group of TOB and fluorescamine in borate buffer, pH 8.5, to give a highly fluorescent derivative which is measured at 469 nm after excitation at 388 nm. The fluorescence intensity was directly proportional to the concentration over the range 300-1500 ng/mL, with a limit of detection of 65 ng/mL and limit of quantitation of 215 ng/mL. All variables were investigated to optimize the reaction conditions. The method was validated according to International Conference on Harmonization guidelines in terms of specificity, linearity, limit of detection, limit of quantification, accuracy, precision and robustness. Good recoveries were obtained ranging from 97.4 to 100.64%, indicating that no interference was observed from concomitants usually present in pharmaceutical dosage forms. The method was successfully, applied for the analysis of the drug substance in its pharmaceutical preparations and spiked serum samples.
一种简单、灵敏和选择性的荧光分光光度法已被开发用于测定人血清和药物制剂中的妥布霉素（TOB）。该方法基于 TOB 的伯氨基与硼酸缓冲液（pH 8.5）中的荧光胺之间的反应，生成一种高度荧光的衍生物，在 388nm 激发下在 469nm 处测量其荧光强度。荧光强度与浓度在 300-1500ng/mL 范围内呈正比，检测限为 65ng/mL，定量限为 215ng/mL。对所有变量进行了研究，以优化反应条件。该方法已根据国际协调会议指南进行了特异性、线性、检测限、定量限、准确度、精密度和稳定性的验证。从 97.4%到 100.64%的回收率良好，表明通常存在于药物制剂中的伴随物没有观察到干扰。该方法成功地应用于药物制剂中药物物质和加标血清样品的分析。
