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首页> 外文期刊>Environmental Science & Technology >Molecular Size Distribution of Fluorophores in Aquatic Natural Organic Matter: Application of HPSEC with Multi-Wavelength Absorption and Fluorescence Detection Following LPSEC-PAGE Fractionation
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Molecular Size Distribution of Fluorophores in Aquatic Natural Organic Matter: Application of HPSEC with Multi-Wavelength Absorption and Fluorescence Detection Following LPSEC-PAGE Fractionation

机译：水生天然有机物中荧光团的分子大小分布：LPSEC-PAGE分馏后具有多波长吸收和荧光检测的HPSEC的应用

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摘要

Analytical high performance size exclusion liquid chromatography (HPSEC) with multiwavelength absorbance and fluorescence detections was used for the analysis of molecular size distribution and optical properties of dissolved natural organic matter. Experiments were conducted on Suwannee River organic matter (SRNOM) and its fractions A, B, C+D preliminary obtained by combination of preparative low pressure size exclusion chromatography and polyacrylamide gel electrophoresis (LPSEC-PAGE) and purified by dialysis on membrane with nominal cutoff 10 kDa, the fractions molecular size varied in order A > B > C + D > 10 kDa. The multistep fractionation of SRNOM enabled the size-separation of at least five types of humic-like fluorophores within NOM showing emission maxima at 465, 450, 435, 420, and 405 nm. The decrease of the humic-like emission maxima paralleled the decrease of the nominal molecular size of fluorescent SRNOM. The protein-like fluorescence was split into tyrosine-like and tryptophan-like fluorophores and only detected in fractions A and B. This work provides new data on the optical properties of size-fractionated NOM, which consistent with the formation of supramolecular NOM assemblies, likely controlled by association of low-molecular size components. It is clearly observed for the high molecular size fraction A, containing free amino acids or short peptides. The combination of several different fractionation procedures is very useful for obtaining less complex NOM compounds and understanding the NOM function in the environment.
机译：具有多波长吸收和荧光检测的分析型高性能尺寸排阻液相色谱（HPSEC）用于分析溶解的天然有机物的分子大小分布和光学性质。对Suwannee River有机物（SRNOM）及其级分A，B，C + D进行了初步制备，该级分是通过制备型低压尺寸排阻色谱法和聚丙烯酰胺凝胶电泳（LPSEC-PAGE）组合得到的，并通过在膜上渗析并按名义截止值进行纯化10 kDa时，各部分的分子大小按A> B> C + D> 10 kDa的顺序变化。 SRNOM的多步分级能够实现NOM内至少五种类型的腐殖质类荧光团的大小分离，在465、450、435、420和405 nm处显示最大发射。腐殖质发射最大值的减少与荧光SRNOM标称分子大小的减少平行。蛋白质样荧光被分为酪氨酸样和色氨酸样荧光团，仅在级分A和B中被检测到。这项工作提供了有关大小分级NOM光学性质的新数据，这与超分子NOM组装体的形成一致，可能是由低分子组分的结合所控制的。对于包含游离氨基酸或短肽的高分子量级分A，可以清楚地观察到。几种不同的分馏程序的组合对于获得不太复杂的NOM化合物和了解环境中的NOM功能非常有用。

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  • 来源
    《Environmental Science & Technology》 |2018年第9期|5287-5295|共9页
  • 作者

    Oleg A. Trubetskoj; Claire Richard; Guillaume Voyard; Victor V. Marchenkov; Olga E. Trubetskaya;

  • 作者单位

    Institute of Basic Biological Problems, Russian Academy of Sciences, 142290, Pushchino, Moscow Region Russia;

    Clermont Université, Université Blaise Pascal, Institut de Chimie de Clermont-Ferrand, Equipe Photochimie, BP 10448, F-63000 Clermont-Ferrand, France,CNRS, UMR 6296, ICCF, F-63171 Aubiere, France;

    Clermont Université, Université Blaise Pascal, Institut de Chimie de Clermont-Ferrand, Equipe Photochimie, BP 10448, F-63000 Clermont-Ferrand, France,CNRS, UMR 6296, ICCF, F-63171 Aubiere, France;

    Institute of Protein Research, Russian Academy of Sciences, 142290 Pushchino, Moscow Region Russia;

    Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 142290, Pushchino, Moscow Region Russia;

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