4-Nitrobenzaldehyde is the synthetic raw material and an important photodegradation product of chloramphenicol. With a structural "alert" of human genotoxic potential and reported mutagenicity, this compound should be controlled in drug substances as a potential genotoxic impurity. However, current analysis methods require complex pre-treatment processes and/or lack sufficient specificity and sensitivity. Nitrophenylhydrazine is a common carbonyl derivatization reagent used to determine the residual aromatic aldehydes in drug samples. In the present study, we report an unexpected advantage of 3-nitrophenylhydrazine hydrochloride as a derivatization reagent in the derivatization high-performance liquid chromatography-ultraviolet detection method to determine 4-nitrobenzaldehyde in chloramphenicol samples. Compared with other nitro-substituted phenylhydrazines, 3-nitrophenylhydrazine hydrochloride can minimize drug matrix and derivatization reagent interferences, since the maximum absorption wavelength of its derivative is significantly red-shifted to 397 nm. The derivatization conditions have been optimized in terms of reaction efficiency, including reaction temperature, time, and diluting solvent, through a design of experiments. As a result, after reaction with 500 μg mL-1 of 3-nitrophenylhydrazine hydrochloride in acetonitrile-water (70:30, v/v) at 60 °C for 30 min, the developed HPLC method could be used to determine 4-nitrobenzaldehyde with a limit of detection of 0.009 μg mL-1. The method was then validated and applied for the determination of residual 4-nitrobenzaldehyde in chloramphenicol and its eye-drop samples.

中文翻译：

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3-硝基苯肼衍生化后，通过HPLC / UV / Vis检测，通过HPLC测定氯霉素及其药物制剂中残留的4-硝基苯甲醛。

4-硝基苯甲醛是氯霉素的合成原料和重要的光降解产物。具有人类遗传毒性潜力的结构“警报”和已报道的诱变性，应在药物中控制该化合物作为潜在的遗传毒性杂质。然而，当前的分析方法需要复杂的预处理过程和/或缺乏足够的特异性和敏感性。硝基苯肼是一种常见的羰基衍生试剂，用于确定药物样品中的残留芳香醛。在本研究中，我们报告了在衍生化高效液相色谱-紫外检测方法中测定氯霉素样品中的4-硝基苯甲醛的方法，将3-硝基苯肼盐酸盐作为衍生试剂具有出乎意料的优势。与其他硝基取代的苯肼相比，3-硝基苯肼的盐酸盐可将药物基质和衍生化试剂的干扰降到最低，因为其衍生物的最大吸收波长显着红移至397 nm。通过实验设计，已在反应效率方面优化了衍生化条件，包括反应温度，时间和稀释溶剂。结果，在60°C下与500μgmL-1的3-硝基苯肼盐酸盐在乙腈-水（70:30，v / v）中反应30分钟后，可以使用发达的HPLC方法测定4-硝基苯甲醛检出限为0.009μgmL-1。然后对该方法进行了验证，并将其用于测定氯霉素及其滴眼液样品中的残留4-硝基苯甲醛。3-硝基苯肼盐酸盐可以最大程度地减少药物基质和衍生试剂的干扰，因为其衍生物的最大吸收波长显着红移至397 nm。通过实验设计，已在反应效率方面优化了衍生化条件，包括反应温度，时间和稀释溶剂。结果，在60°C下与500μgmL-1的3-硝基苯肼盐酸盐在乙腈-水（70:30，v / v）中反应30分钟后，可以使用发达的HPLC方法测定4-硝基苯甲醛检出限为0.009μgmL-1。然后对该方法进行了验证，并将其用于测定氯霉素及其滴眼液样品中的残留4-硝基苯甲醛。3-硝基苯肼盐酸盐可以最大程度地减少药物基质和衍生试剂的干扰，因为其衍生物的最大吸收波长显着红移至397 nm。通过实验设计，已在反应效率方面优化了衍生化条件，包括反应温度，时间和稀释溶剂。结果，在60°C下与500μgmL-1的3-硝基苯肼盐酸盐在乙腈-水（70:30，v / v）中反应30分钟后，可以使用发达的HPLC方法测定4-硝基苯甲醛检出限为0.009μgmL-1。然后对该方法进行了验证，并将其用于测定氯霉素及其滴眼液样品中的残留4-硝基苯甲醛。