Dipartimento di Chimica e Tecnologie Chimiche, Università della Calabria, Via P. Bucci Cubo 12/C, I-87030, Arcavacata di Rende, CS, Italy; CNR-Institute of Atmospheric Pollution Research, Division of Rende, c/o UNICAL-Polifunzionale, I-87036, Arcavacata di Rende, CS, Italy.
Dipartimento di Chimica e Tecnologie Chimiche, Università della Calabria, Via P. Bucci Cubo 12/C, I-87030, Arcavacata di Rende, CS, Italy.
J Chromatogr A. 2018 May 11;1549:1-13. doi: 10.1016/j.chroma.2018.03.034. Epub 2018 Mar 19.
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., N-Acetylspermidine, N-Acetylspermidine, and N-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.
多胺是具有低分子量的脂族胺，被广泛认为是早期诊断和治疗的最重要的癌症生物标志物之一。本文工作的目标是开发一种快速简便的方法，用于定量测定人尿中的游离多胺（即腐胺、尸胺、亚精胺、精胺）和 N-单乙酰化多胺（即 N-乙酰基精胺、N-乙酰基亚精胺和 N-乙酰基精胺）。与丙酰氯甲酸酯的初步衍生化结合固相微萃取（SPME），可以实现一种简单且自动化的方案，涉及最小的样品处理，不消耗有机溶剂。以单变量模式评估了分析物对五种商业 SPME 涂层的亲和力，以二乙烯基苯/羧基/聚二甲基硅氧烷纤维的分析物提取效果最佳。通过实验设计的多元方法，特别是使用中心组合设计（CCD），优化了影响 SPME 分析性能的变量。从响应值的角度来看，最佳工作条件为：萃取温度 40°C，萃取时间 15min，不添加 NaCl。通过气相色谱-三重四极杆质谱（GC-QqQ-MS）在选择反应监测（SRM）采集模式下进行分析。该方法按照美国食品和药物管理局（FDA）发布的指南进行了验证。在线性、灵敏度（LOQs 在 0.01 至 0.1μg/mL 之间）、基质效应（68-121%）、准确性和精密度（日内值在-24%至+16%之间，范围在 3.3-28.4%之间）方面取得了令人满意的结果，使所提出的方案适合用于定量测定尿液样本中的这些重要生物标志物。
