Combining electrophoresis with detection under ultraviolet light and multiple ultrafiltration for isolation of humic fluorescence fractions
| Autor: | Oleg Trubetskoj, Lubov Shaloiko, Christian Coelho, I. I. Proskuryakov, Olga Trubetskaya, Dmitrii V. Demin, Victor V. Marchenkov |
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| Rok vydání: | 2010 |
| Předmět: |
Fluorophore
Chromatography Ultraviolet Rays Size-exclusion chromatography Analytical chemistry Ultrafiltration Fluorescence spectrometry Biochemistry Fluorescence Analytical Chemistry Gel permeation chromatography chemistry.chemical_compound chemistry Sephadex Ultraviolet light Environmental Chemistry Urea Electrophoresis Polyacrylamide Gel Spectroscopy Humic Substances |
| Zdroj: | Analytica chimica acta. 690(2) |
| ISSN: | 1873-4324 |
| Popis: | Polyacrylamide gel electrophoresis of chernozem soil humic acids (HAs) followed by observation under UV (312 nm) excitation light reveals new low molecular weight (MW) fluorescent fractions. Ultrafiltration of HAs sample in 7 M urea on a membrane of low nominal MW retention (NMWR, 5 kDa) was repetitively used for separation of fluorescent and non-fluorescent species. Thirty ultrafiltrates and the final retentate R were obtained. Fluorescence maxima of separate ultrafiltrates were different and non-monotonously changed in the range of 475–505 nm. Fluorescence maxima of less than 490 nm were detected only in the four first utrafiltrates. For further physical–chemical analyses all utrafiltrates were combined into a fraction called UF < 5 (NMW < 5 kDa). Retentate R demonstrated very weak fluorescence under 270 nm excitation, while fluorescence intensity of UF < 5 was about six times higher than of the bulk HAs. Fraction UF < 5 was further ultrafiltrated on membranes of MNWR 3 kDa and 1 kDa, yielding three subfractions UF3-5, UF1-3 and UF < 1 with NMW 3–5 kDa, 1–3 kDa and |
| Databáze: | OpenAIRE |
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