Fluorous-paired derivatization approach towards highly sensitive and accurate determination of long chain unsaturated fatty acids by liquid chromatography-tandem mass spectrometry.
Fluorous-paired derivatization approach towards highly sensitive and accurate determination of long chain unsaturated fatty acids by liquid chromatography-tandem mass spectrometry.
复制标题
DOI：
10.1016/j.aca.2020.09.052
复制
发表时间：
2020-11
影响因子：
6.2
通讯作者：
Jia-Yi Zheng;Ying-Ying Jin-Ying;Zi-Qi Shi;Jian-Liang Zhou;Li-Fang Liu;G. Xin
中科院分区：
文献类型：
--
作者：
Jia-Yi Zheng;Ying-Ying Jin-Ying;Zi-Qi Shi;Jian-Liang Zhou;Li-Fang Liu;G. Xin
长链不饱和脂肪酸（LCUFA）正在成为炎症及其解决的关键因素。因此，生物样品中LCUFA的灵敏和准确测量在疾病诊断和预后中具有重要价值。本研究采用一对含氟试剂，即3-（全氟辛基）-丙胺（PFPA）和2-（全氟辛基）-乙胺（PFEA），建立了LCUFA的UPLC-MS/MS定量的氟衍生化方法。该方法将生物样品中的LCUFA用PFPA进行全氟烷基化，并特异性地保留在氟相液相色谱柱上，大大降低了基质干扰引起的定量偏差。此外，引入了PFEA标记的LCUFA标准品作为一对一的内标，以最大限度地确保无偏倚的结果。应用该方法能够可靠地测定8种典型的LCUFA，具有高灵敏度（LLOQ范围从30阿莫尔到6.25 fmol）和低基质干扰（几乎小于10%）。这样的高灵敏度可以方便地测定小体积和低浓度的生物样品。在OVA诱导的哮喘小鼠中监测了这些靶向LCUFA的进一步代谢特征，仅需要5 μL血清样本。我们的研究结果表明，哮喘发作导致显着的干扰不仅在浓度，而且在这些LCUFA之间的比例。鉴于灵敏度和准确性方面的有利优势，本氟配对衍生化方法有望成为剖析LCUFA的生理和临床意义的新途径，从而阐明与其干扰相关的疾病的管理。
Long chain unsaturated fatty acids (LCUFAs) are emerging as critical contributors to inflammation and its resolution. Sensitive and accurate measurement of LCUFAs in biological samples is thus of great value in disease diagnosis and prognosis. In this work, a fluorous-derivatization approach for UPLC-MS/MS quantification of LCUFAs was developed by employing a pair of fluorous reagents, namely 3-(perfluorooctyl)-propylamine (PFPA) and 2-(perfluorooctyl)-ethylamine (PFEA). With this method, the LCUFAs in biological samples were perfluoroalkylated with PFPA and specifically retained on a fluorous-phase LC column, which largely reduced matrix interferences-induced quantitation deviation. Moreover, PFEA-labeled LCUFAs standards were introduced as one-to-one internal standards to farthest ensure unbiased results. Application of the proposed method enabled a reliable determination of eight typical LCUFAs with high sensitivity (LLOQ ranged from 30 amol to 6.25 fmol) and low matrix interferences (almost less than 10%). Such a high sensitivity could facilitate the determination of small-volume and low-concentration bio-samples. Further metabolic characterization of these targeted LCUFAs was monitored in OVA-induce asthma mice, requiring only 5 μL serum sample. Our results showed that asthmatic attack led to significant disturbances not only in the concentrations but also in the ratio among these LCUFAs. In view of the favorable advantages in sensitivity and accuracy, the present fluorous-paired derivatization approach will be expected to serve as a new avenue for dissecting the physiological and clinical implications of LCUFAs, thereby shedding light on the management of diseases related to their disturbances.