Leicester Institute for Structural and Chemical Biology and School of Chemistry, University of Leicester, Henry Wellcome Building, Lancaster Road, Leicester, LE1 7RH, UK.
Space Park Leicester, University of Leicester, 92 Corporation Road, Leicester, LE4 5SP, UK.
Sci Rep. 2024 Sep 4;14(1):20621. doi: 10.1038/s41598-024-71271-z.
Formaldehyde (HCHO) is a human toxin that is both a pollutant and endogenous metabolite. HCHO concentrations in human biological samples are reported in the micromolar range; however, accurate quantification is compromised by a paucity of sensitive analysis methods. To address this issue, we previously reported a novel SPME-GC-MS-based HCHO detection method using cysteamine as an HCHO scavenger. This method showed cysteamine to be a more efficient scavenger than the widely used O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine, and enabled detection of aqueous HCHO in the nanomolar range and quantification in the micromolar range. However, quantification in this range required immersive extraction of the HCHO-derived thiazolidine, while a high background signal was also observed. Following on from these studies, we now report an optimised head-space extraction SPME-GC-MS method using cysteamine, which provides similarly sensitive HCHO quantification to the immersive method but avoids extensive wash steps and is therefore more amenable to screening applications. However, high background HCHO levels were still observed A Complementary GC-MS analyses using a 2-aza-Cope-based HCHO scavenger also revealed high background HCHO levels; therefore, the combined results suggest that HCHO exists in high (i.e. micromolar) concentration in aqueous samples that precludes accurate quantification below the micromolar range. This observation has important implications for ongoing HCHO quantification studies in water, including in biological samples.
甲醛(HCHO)是一种人类毒素,既是污染物又是内源性代谢物。人体生物样本中的 HCHO 浓度报告在微摩尔范围内;然而,由于缺乏敏感的分析方法,准确的定量受到了限制。为了解决这个问题,我们之前报道了一种使用半胱氨酸作为 HCHO 清除剂的新型 SPME-GC-MS 基 HCHO 检测方法。该方法表明半胱氨酸比广泛使用的 O-(2,3,4,5,6-五氟苄基)羟胺更有效地清除 HCHO,并且能够在纳摩尔范围内检测到水溶液中的 HCHO,并在微摩尔范围内进行定量。然而,在此范围内进行定量需要沉浸式提取 HCHO 衍生的噻唑烷,同时也观察到高背景信号。在这些研究的基础上,我们现在报告了一种使用半胱氨酸优化的顶空萃取 SPME-GC-MS 方法,该方法提供了与沉浸式方法类似的敏感 HCHO 定量,但避免了广泛的洗涤步骤,因此更适合筛选应用。然而,仍然观察到高背景 HCHO 水平。使用基于 2-氮杂-Cope 的 HCHO 清除剂的补充 GC-MS 分析也揭示了高背景 HCHO 水平;因此,综合结果表明,在水溶液中 HCHO 以高浓度(即微摩尔)存在,这排除了在微摩尔以下范围内进行准确定量的可能性。这一观察结果对水,包括生物样本中的 HCHO 定量研究具有重要意义。