Fanti F, Merola C, Vremere A, Oliva E, Perugini M, Amorena M, Compagnone D, Sergi M
University of Teramo, Faculty of Bioscience and Technology for Food, Agriculture and Environment, 64100 TE, Italy.
University of Teramo, Faculty of Bioscience and Technology for Food, Agriculture and Environment, 64100 TE, Italy.
Talanta. 2020 Dec 1;220:121393. doi: 10.1016/j.talanta.2020.121393. Epub 2020 Jul 11.
Zebrafish is an in vivo model used in toxicology to estimate the effects of xenobiotics and their teratogenic consequences. The knowledge of the oxysterols profile in zebrafish, during early embryonic stages, provides important information on the role and biological function of these molecules. This work reports the development and validation of a LC-MS/MS method for the determination of 7 different oxysterols in zebrafish embryos. Sample was treated with a combination of liquid/liquid extraction (LLE) followed by micro solid phase extraction (μSPE) clean-up in order to remove matrix interference and obtain a suitable enrichment factor of the analytes. The method was validated on 2 different embryos growing stages, 3-4 and 24 h post fertilization (hpf), as slight differences in terms of recovery and matrix effect were shown. The validation results provided good accuracy (bias ≤17%; 20% at LOQ) and repeatability (≤15%; ≤19% at LOQ), with low LOQs in the range 22 and 65 pg on 100 embryos sample, without any analyte derivatization, demonstrating the suitability of this analytical method as a useful tool to understand the correlation between oxysterols profile and developmental abnormalities induced by xenobiotic exposure.
斑马鱼是一种用于毒理学研究的体内模型，用于评估外源化合物的影响及其致畸后果。了解斑马鱼早期胚胎阶段的氧甾醇谱，可为这些分子的作用和生物学功能提供重要信息。本文报道了一种用于测定斑马鱼胚胎中7种不同氧甾醇的液相色谱-串联质谱（LC-MS/MS）方法的开发与验证。样品先经过液液萃取（LLE），然后进行微固相萃取（μSPE）净化处理，以去除基质干扰并获得合适的分析物富集因子。该方法在受精后3-4小时和24小时这两个不同的胚胎发育阶段进行了验证，因为在回收率和基质效应方面显示出了细微差异。验证结果显示出良好的准确性（偏差≤17%；定量限处为20%）和重复性（≤15%；定量限处为≤19%），在100个胚胎样品上的定量限低至22至65 pg，无需对任何分析物进行衍生化处理，证明了该分析方法作为了解氧甾醇谱与外源化合物暴露诱导的发育异常之间相关性的有用工具的适用性。
