title: Evaluation of Suwannee River NOM electrophoretic fractions by RP-HPLC with online absorbance and fluorescence detection.
abstract: Reversed-phase high-performance liquid chromatography (RP-HPLC) with online absorbance and fluorescence detections was used for the evaluation of Suwannee River natural organic matter (SRNOM) and its fractions A, B, and C+D, obtained by conventional size exclusion chromatography-polyacrylamide gel electrophoresis (SEC-PAGE) setup, for which the electrophoretic mobility (EM) and the absorptivity varied in the order C+D > B > A, and the molecular size (MS) in the opposite order. Analysis of SRNOM and its fractions in part of their relative irreversible adsorption on C18-column and relative distribution of eluted from the column matter on hydrophobic and hydrophilic peaks showed that hydrophobicity of fractions decreased in order: A > B > C+D. The online fluorescence detection showed that SRNOM and its fractions contained at least three groups of humic substances (HS)-like fluorophores with emission maxima at 435, 455-465, and 455/420 nm and two protein-like fluorophores with emission maxima at around 300 and 340 nm. The HS-like fluorophore with emission maximum at 435 nm was located in the hydrophilic peak in all the samples. Those with maxima at 455-465 nm were detected in hydrophobic peaks of fractions A and B. SEC-PAGE setup followed by RP-HPLC allowed us to develop new approach of SRNOM separation on less heterogeneous compounds mixture for their further study and structural identification.
authors: Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Pushchino, Moscow region, Russia, 142290.
authors: Olga E
authors: OE
authors: Trubetskaya
authors: Claire
authors: C
authors: Richard
authors: Oleg A
authors: OA
authors: Trubetskoj
mesh_terms: D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D056148:Chromatography, Reverse-Phase; D004591:Electrophoresis, Polyacrylamide Gel; D004784:Environmental Monitoring / Q000379:methods*; D005453:Fluorescence; D005845:Georgia; D006812:Humic Substances / Q000032:analysis*; D057927:Hydrophobic and Hydrophilic Interactions; D011506:Proteins; D045483:Rivers / Q000737:chemistry*; D014873:Water Pollutants / Q000032:analysis*
chemical_list: D006812:Humic Substances; D011506:Proteins; D014873:Water Pollutants