A Fluorescent-Based HPLC Assay Using 4-Chloro-7-nitrobenzo-2-oxa-1, 3-diazole as Derivatization Agent for the Determination of Iron Bioavailability to Red Tide Phytoplankton

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• 作者：H. Hasegawa (1)
  K. Nakagawa (2)
  M. Azizur Rahman (3)
  M. Takemura (2)
  T. Maki (1)
  K. Naito (4)
  M. Mamunur Rahman (2) (5)
  
  1. Institute of Science and Technology ; Kanazawa University ; Kakuma ; Kanazawa ; 920-1161 ; Japan
  2. Graduate School of Natural Science and Technology ; Kanazawa University ; Kakuma ; Kanazawa ; 920-1192 ; Japan
  3. Centre for Environmental Sustainability ; School of the Environment ; University of Technology Sydney ; PO Box 123 ; Broadway ; Sydney ; NSW ; 2007 ; Australia
  4. Faculty of Life and Environmental Sciences ; Prefectural University of Hiroshima ; Shobara ; Hiroshima ; 727-0023 ; Japan
  5. Bangladesh Rice Research Institute (BRRI) ; Gazipur-1701 ; Dhaka ; Bangladesh
  
• 关键词：Fluorescent ; based HPLC assay ; Marine phytoplankton ; Red tide ; Iron bioavailability ; NBD ; C1
• 刊名：Chromatographia
• 出版年：2015
• 出版时间：January 2015
• 年：2015
• 卷：78
• 期：1-2
• 页码：65-72
• 全文大小：901 KB
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• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Analytical Chemistry
  Organic Chemistry
  Pharmacy
  Biochemistry
  Plant Sciences
  Measurement Science and Instrumentation
  
• 出版者：Vieweg Verlag
• ISSN：1612-1112

文摘

A new fluorescent-based high-performance liquid chromatography (HPLC) assay using 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole (NBD-C1) was employed to determine iron (Fe) bioavailability to red tide phytoplankton in seawater. After growing four red tide species (Prymnesium parvum, Heterosigma akashiwo, Eutreptiella gymnastica, and Oltmannsiellopsis viridis) in f/2 artificial seawater under different Fe conditions, soluble extracts of the phytoplankton were derivatized using different fluorescent reagents (NBD-C1, 4-fluoro-7-nitrobenzo-2-oxa-1,3-diazole; NBD-F, fluorescamine, and ortho-phthalaldehyde; OPA) followed by HPLC assay. Among the four fluorescent reagents, NBD-C1 was most effective for derivatizing the phytoplankton extracts which would consist of proteins and peptides. HPLC chromatograms of the NBD-derivatized extracts showed gradual changes (decrease/increase) of six peaks for different Fe conditions. Four of the peaks decreased, while two peaks increased with the increase of Fe concentrations in the culture medium. Considering the consistency and sensitivity of chromatogram peaks E and A to different Fe, phosphate and nitrate conditions for all phytoplankton studied, the ratio of these two peaks (I E/A ) has been proposed as the indicator of Fe bioavailability to red tide phytoplankton.