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Two-stage bile preparation with acetone for recovery of fluorescent aromatic compounds (FACs)

机译：用丙酮分两步制备胆汁以回收荧光芳香化合物（FAC）

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摘要

In this study we sought to optimize recovery of fluorescent aromatic compounds (FACs) from the bile of African catfish (Clarias gariepinus) injected with 10mg/kg benzo[a]pyrene (BaP). Fractions of pooled bile were hydrolyzed, combined with ten volumes of methanol, ethanol, acetonitrile, or acetone, cen-trifuged and supernatants were analyzed by high-performance liquid chromatography with fluorescent detection (HPLC/FL). As well, to test whether FACs were being lost in solids from the centrifugation, pellets were resuspended, hydrolyzed and mixed with six volumes of the organic solvent that produced best FAC recovery from the supernatant, and subjected to HPLC/FL, Highest FAC concentrations were obtained with 2000 μJ and 1250 μ.1 acetone for supernatants and resuspended pellets respectively. FACs concentrations were negatively correlated with biliary protein content but were unaffected by addition of bovine serum albumin (BSA) followed by no incubation indicating that the presence of proteins in the biliary mixture does not simply interfere with detection of FACs. In another experiment, efficiency of acetone addition was compared to two different liquid-liquid extractions (L-LEs). Acetone additions provided significantly higher biliary FACs than the L-LE methods. The new two-stage bile preparation with acetone is an efficient, inexpensive and easily performed method.
机译：在这项研究中，我们试图优化注射10mg / kg苯并[a] py（BaP）的非洲cat鱼（Clarias gariepinus）胆汁中荧光芳香化合物（FAC）的回收率。水解合并的胆汁级分，与十倍体积的甲醇，乙醇，乙腈或丙酮混合，离心，并通过高效液相色谱-荧光检测（HPLC / FL）分析上清液。同样，为了测试离心分离过程中FAC是否在固体中丢失，将沉淀重新悬浮，水解并与六种体积的有机溶剂混合，这些有机溶剂可从上清液中获得最佳的FAC回收率，并进行HPLC / FL处理，FAC浓度最高。用2000μJ和1250μ.1丙酮分别获得上清液和重悬的沉淀。 FACs浓度与胆汁蛋白质含量呈负相关，但不受添加牛血清白蛋白（BSA）的影响，随后未孵育，表明胆汁混合物中蛋白质的存在并不简单地干扰FACs的检测。在另一个实验中，将丙酮的添加效率与两种不同的液-液萃取（L-LE）进行了比较。丙酮添加比L-LE方法提供更高的胆汁FAC。新型的两步丙酮胆汁制备方法是一种高效，廉价且易于执行的方法。

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  • 来源
    《Journal of Hazardous Materials》 |2012年第15期|p.84-93|共10页
  • 作者

    Ali Karami; Mohd A. Syed; Annie Christianus; Kristine L. Willett; Jeffrey R. Mazzeo; Simon C. Courtenay;

  • 作者单位

    Department of Aquaculture, Faculty of Agriculture, Universiti Putra Malaysia, 43400 Selangor, Malaysia;

    Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Selangor, Malaysia;

    Department of Aquaculture, Faculty of Agriculture, Universiti Putra Malaysia, 43400 Selangor, Malaysia;

    Department of Pharmacology and Environmental Toxicology Research Program, University of Mississippi, University, MS 38677, USA;

    Pharmaceutical Business Operations, Waters Corporation, 34 Maple Street, Milford, MA 01757, USA;

    Fisheries and Oceans Canada at the Canadian Rivers Institute, Department of Biology, University of New Brunswick, Fredericton, New Brunswick, Canada;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    fish bile; fluorescent aromatic compounds (FACs); acetone; two-stage preparation; protein;

    机译：鱼胆荧光芳香族化合物（FAC）;丙酮;两阶段准备;蛋白;

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