Journal
ANALYTICA CHIMICA ACTA
Volume 408, Issue 1-2, Pages 299-307Publisher
ELSEVIER
DOI: 10.1016/S0003-2670(99)00880-6
Keywords
water analysis; thiols; phytochelatin; soft metals; fluorescence detection
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A sensitive and reliable reversed-phase high-performance liquid chromatography (HPLC) method has been developed to analyze naturally occurring low-molecular weight thiols using pre-column derivatization with ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulfonate (SBD-F) and fluorescence detection in a gradient elution. The derivatization procedure and separation conditions were optimized by careful control of the reaction conditions in terms of time, temperature, pH and reducing agent. A linear calibration in the nM range was obtained for all the standard compounds, allowing measurement of thiols in natural water samples. Results show that the reducing agent tri-n-butylphosphine (TBP) not only acts as an anti-oxidant/disulfide cleaving agent, but also enables the SBD-F to react with thiols complexed by soft metals. This is the first study reported using the SBD-F labeling technique to measure phytochelatin concentrations. The methodology has been successfully applied to estuarine water samples from Galveston Bay. (C) 2000 Elsevier Science B.V. All rights reserved.
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