Red or processed meat rich diets have been shown to be associated with an elevated risk of colorectal cancer (CRC). One major hypothesis involves dietary heme iron which induces lipid peroxidation. The quantification of the resulting reactive aldehydes (e.g. HNE and HHE) in the colon lumen is therefore of great concern since these compounds are known for their cytotoxic and genotoxic properties. UHPLC-ESI-MS/MS method has been developed and validated for HNE and HHE quantification in rat faeces. Samples were derivatised using a brominated reagent (BBHA) in presence of pre-synthesized deuterated internal standards (HNE-d11/HHE-d5), extracted by solid phase extraction, and then analysed by LC-positive ESI-MS/MS (MRM) on a TSQ Vantage mass spectrometer. The use of BBHA allowed the efficient stabilisation of the unstable and reactive hydroxy-alkenals HNE and HHE. The MRM method allowed selective detection of HNE and HHE on the basis of characteristic transitions monitored from both the 79 and 81 bromine isotopic peaks. This method was validated according to the European Medicines Agency (EMEA) guidelines, by determining selectivity, sensitivity, linearity, carry-over effect, recovery, matrix effect, repeatability, trueness and intermediate precision. The performance of the method enabled the quantification of HNE and HHE in concentrations 0.10–0.15 μM in faecal water. Results are presented on the application to the quantification of HNE and HHE in different faecal waters obtained from faeces of rats fed diets with various fatty acid compositions thus corresponding to different pro-oxidative features.

已经证明，富含红色或加工肉类的饮食与大肠癌（CRC）的风险升高有关。一种主要假说涉及饮食血红素铁，其引起脂质过氧化。由于在结肠腔中产生的反应性醛（例如，HNE和HHE）的数量众所周知，因为这些化合物具有细胞毒性和基因毒性特性，因此这些问题引起了人们的极大关注。已经开发了UHPLC-ESI-MS / MS方法并验证了大鼠粪便中的HNE和HHE定量。在预先合成的氘代内标（HNE-d11 / HHE-d5）存在下，使用溴化试剂（BBHA）衍生样品，通过固相萃取进行萃取，然后通过LC阳性ESI-MS / MS（MRM）进行分析在TSQ Vantage质谱仪上 BBHA的使用可有效稳定不稳定和反应性的羟基烯烃HNE和HHE。MRM方法可以根据从79个和81个溴同位素峰监测到的特征跃迁选择性检测HNE和HHE。通过确定选择性，灵敏度，线性，残留效应，回收率，基质效应，可重复性，真实性和中间精度，该方法已根据欧洲药品管理局（EMEA）指南进行了验证。该方法的性能使粪便中HNE和HHE的浓度在0.10–0.15μM范围内得以定量。