Polyamines are aliphatic amines with low molecular weight that are widely recognized as one of the most important cancer biomarkers for early diagnosis and treatment. The goal of the work herein presented is the development of a rapid and simple method for the quantification of free polyamines (i.e., putrescine, cadaverine, spermidine, spermine) and N-monoacetylated polyamines (i.e., N¹-Acetylspermidine, N⁸-Acetylspermidine, and N¹-Acetylspermine) in human urine. A preliminary derivatization with propyl chloroformate combined with the use of solid phase microextraction (SPME) allowed for an easy and automatable protocol involving minimal sample handling and no consumption of organic solvents. The affinity of the analytes toward five commercial SPME coatings was evaluated in univariate mode, and the best result in terms of analyte extraction was achieved using the divinylbenzene/carboxen/polydimethylsiloxane fiber. The variables affecting the performance of SPME analysis were optimized by the multivariate approach of experimental design and, in particular, using a central composite design (CCD). The optimal working conditions in terms of response values are the following: extraction temperature 40 °C, extraction time of 15 min and no addition of NaCl. Analyses were carried out by gas chromatography-triple quadrupole mass spectrometry (GC-QqQ-MS) in selected reaction monitoring (SRM) acquisition mode. The developed method was validated according to the guidelines issued by the Food and Drug Administration (FDA). The satisfactory performances reached in terms of linearity, sensitivity (LOQs between 0.01 and 0.1 μg/mL), matrix effect (68–121%), accuracy, and precision (inter-day values between −24% and +16% and in the range 3.3–28.4%, respectively) make the proposed protocol suitable to be adopted for quantification of these important biomarkers in urine samples.

多胺是低分子量的脂肪胺，被广泛认为是早期诊断和治疗的最重要的癌症生物标记之一。本文所呈现的工作的目的是快速和简单的方法为自由多胺（即，腐胺，尸胺，亚精胺，精胺）和量化的发展Ñ -monoacetylated多胺（即，Ñ ¹ -乙酰，Ñ ⁸ -乙酰和N ¹-乙酰精胺）在人类尿液中。用氯甲酸丙酯进行初步衍生化并结合使用固相微萃取（SPME），可实现简单且可自动化的操作流程，包括最少的样品处理且不消耗有机溶剂。以单变量模式评估了分析物对五种市售SPME涂层的亲和力，使用二乙烯基苯/羧酸/聚二甲基硅氧烷纤维获得了分析物提取方面的最佳结果。通过多变量实验设计方法，特别是使用中央复合设计（CCD），优化了影响SPME分析性能的变量。就响应值而言，最佳工作条件如下：萃取温度40°C，萃取时间15分钟，不添加NaCl。通过气相色谱-三重四极杆质谱（GC-QqQ-MS）在选定的反应监测（SRM）采集模式下进行分析。根据美国食品药品监督管理局（FDA）发行的指南对开发的方法进行了验证。在线性，灵敏度（LOQ在0.01和0.1μg/ mL之间），基质效应（68–121％），准确性和精密度（日间值在-24％和+ 16％之间，以及在范围分别为3.3–28.4％）使拟议的方案适合用于尿液样品中这些重要生物标志物的定量分析。根据美国食品药品监督管理局（FDA）发行的指南对开发的方法进行了验证。在线性，灵敏度（LOQ在0.01和0.1μg/ mL之间），基质效应（68–121％），准确性和精密度（日间值在-24％和+ 16％之间，以及在范围分别为3.3–28.4％）使拟议的方案适合用于尿液样品中这些重要生物标志物的定量分析。根据美国食品药品监督管理局（FDA）发行的指南对开发的方法进行了验证。在线性，灵敏度（LOQ在0.01和0.1μg/ mL之间），基质效应（68–121％），准确性和精密度（日间值在-24％和+ 16％之间，以及在范围分别为3.3–28.4％）使拟议的方案适合用于尿液样品中这些重要生物标志物的定量分析。