High Performance Liquid Chromatographic Determination of Acetoacetate by Post-Column Derivatization with p-Nitrobenzene Diazonium Fluoroborate
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概要
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We have applied a color-developing reagent, p-nitrobenzene diazonium fluoroborate (diazo reagent) as a post-column derivatization tool for the specific determination of acetoacetate (AcAc) in high performance liquid chromatography (HPLC). A mobile phase consisting of 50mM KH_2PO_4, 4mM tetra-n-butylammonium phosphate (TBAP) as an ion-pair reagent and 2v/v% methanol, pH 3.5, diazo reagent solution with 0.2% triton X100, and alkaline solution of 1.5 mol/l NaOH were pumped using three independent pumps. Specific color development on-line was monitored at 645 nm. A calibration curve for AcAc standard solution with an injection volume of 20μl showed a good linearity in the range 0.01-2.5 mM with a correlation coefficient of 0.999. For the determination of 3-hydroxybutyrate (3-HOBA), 3-HOBA was converted to AcAc by an enzymatic-coupling method using 3-HOBA dehydrogenase and lactate dehydrogenase. Analytical recoveries of AcAc and 3-HOBA added to serum and urine were satisfactory.
- 公益社団法人日本薬学会の論文
- 2003-04-01
著者
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小林 匡子
東北薬科大学生薬学教室
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Kobayashi Kyoko
Tohoku Pharmaceutical University
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YAMAMOTO Susumu
Department of Hygiene, Faculty of Medicine, Kagoshima University
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SHIMADA Kenji
Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Niigata University of Pharma
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OHTA SHIN
Department of Pharmacy, Shinshu University Hospital
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YAMATO Susumu
Department of Analytical Chemistry, Niigata College of Pharmacy
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KOBAYASHI Kiyomi
Department of Chemistry, Joetsu University of Education
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Ohta S
Department Of Functional Molecular Chemistry Kyoto Pharmaceutical University
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Ohta Shin
Department Of Hospital Pharmacy Shinshu University School Of Medicine
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Shimada K
Department Of Analytical Chemistry Faculty Of Pharmaceutical Sciences Niigata University Of Pharmacy
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Shimada Kenji
Department Of Analytical Chemistry Faculty Of Pharmaceutical Sciences Niigata University Of Pharmacy
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EBARA Kiko
Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Niigata University of Pharma
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Ebara Kiko
Department Of Analytical Chemistry Faculty Of Pharmaceutical Sciences Niigata University Of Pharmacy
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Kobayashi Kiyomi
Department Of Chemistry Joetsu University Of Education
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Yamamoto Susumu
Department Of Analytical Chemistry Faculty Of Pharmaceutical Sciences Niigata University Of Pharmacy
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Ebara Keiko
Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Niigata University of Pharmacy and Applied Life Sciences
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Yamato Susumu
Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Niigata University of Pharmacy and Applied Life Sciences
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Ohta Shin
Department of Functional Molecular Chemistry, Kyoto Pharmaceutical University
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