两种热带淡水鱼胆汁基线激发-发射荧光的变异性

IF 2.6 4区 化学 Q2 BIOCHEMICAL RESEARCH METHODS Journal of Fluorescence Pub Date : 2024-09-12 DOI:10.1007/s10895-024-03871-x
Diego Mora-Solarte, Rodrigo Jimenez, Ivonne Calderón-Delgado, Alvaro Duarte-Ruiz, Markus Brinkmann, Yohana Velasco-Santamaría
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引用次数: 0

摘要

对鱼类胆汁中的污染物代谢物进行定量是监测淡水中异种生物污染的有效方法,因为这些测量直接针对暴露问题。荧光激发-发射矩阵光谱法(EEMS)已被证明是一种具有高度特异性和成本效益的多环芳烃(PAH)和多环芳烃代谢物鉴定和定量技术。EEMS 量化这些化合物的能力在很大程度上取决于胆汁基线荧光(BBF)的强度和可变性。我们发现 Aequidens metae(AME)个体之间以及这些个体与 Piaractus orinoquensis(PIO)个体之间的 BBF 存在很大差异。此外,BBF 也很大,需要超过 1:400 的溶剂稀释才能避免内部过滤效应。我们使用平行因子分析(PARAFAC)对种内和种间的 BBF 变异进行建模。PARAFAC 成功地将 EEMS 集分解为三种荧光团,它们存在于所有样品中,但浓度相差约 3 个数量级。其中一个因子被确定为色氨酸。色氨酸和因子 2 是共变的,在 AME 中的含量比在 PIO 中高得多,而因子 3 在 PIO 中的含量是在 AME 中的 6 倍。此外,在河中直接捕获的 AME 标本中,色氨酸的含量是实验室适应的同类标本的 10 倍。PARAFAC 分解的有效性通过分数与稀释比的正比例关系得到了证实。一个大的内滤光器表明因子 2 与色氨酸一样具有很强的光吸收能力。我们的研究结果表明,在使用 PARAFAC 检测和定量污染物代谢物时,需要加入胆汁基质变量成分。
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Variability of Bile Baseline Excitation-emission Fluorescence of Two Tropical Freshwater Fish Species

The quantification of pollutant metabolites in fish bile is an efficient approach to xenobiotic pollution monitoring in freshwaters since these measurements directly address exposure. Fluorescence excitation-emission matrix spectroscopy (EEMS) has demonstrated to be a highly specific and cost-effective technique for polycyclic aromatic hydrocarbon (PAH) and PAH-metabolite identification and quantification. EEMS ability to quantify these compounds strongly depends on the intensity and variability of the bile baseline fluorescence (BBF). We found large differences in BBF among Aequidens metae (AME) individuals and of these with Piaractus orinoquensis (PIO). Moreover, BBF was large enough that solvent dilutions of over 1:400 were needed to avoid inner filter effects. We used parallel factor analysis (PARAFAC) to model the intra- and inter-species BBF variability. PARAFAC successfully decomposed the EEMS set into three fluorophores present in all samples, although in concentrations spreading over ~ 3 orders of magnitude. One of the factors was identified as tryptophan. Tryptophan and Factor 2 were covariant and much more abundant in AME than in PIO, while Factor 3 was ~ 6 times more abundant in PIO than in AME. Also, tryptophan was ~ 10x more abundant in AME specimens immediately caught in rivers than in their laboratory-adapted peers. The PARAFAC decomposition effectiveness was confirmed by the positive proportionality of scores to dilution ratios. A large inner filter indicates that Factor 2 is as strong a light absorber as tryptophan. Our results stress the need to include bile matrix variable components for the detection and quantification of pollutant metabolites using PARAFAC.

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来源期刊
Journal of Fluorescence
Journal of Fluorescence 化学-分析化学
CiteScore
4.60
自引率
7.40%
发文量
203
审稿时长
5.4 months
期刊介绍: Journal of Fluorescence is an international forum for the publication of peer-reviewed original articles that advance the practice of this established spectroscopic technique. Topics covered include advances in theory/and or data analysis, studies of the photophysics of aromatic molecules, solvent, and environmental effects, development of stationary or time-resolved measurements, advances in fluorescence microscopy, imaging, photobleaching/recovery measurements, and/or phosphorescence for studies of cell biology, chemical biology and the advanced uses of fluorescence in flow cytometry/analysis, immunology, high throughput screening/drug discovery, DNA sequencing/arrays, genomics and proteomics. Typical applications might include studies of macromolecular dynamics and conformation, intracellular chemistry, and gene expression. The journal also publishes papers that describe the synthesis and characterization of new fluorophores, particularly those displaying unique sensitivities and/or optical properties. In addition to original articles, the Journal also publishes reviews, rapid communications, short communications, letters to the editor, topical news articles, and technical and design notes.
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