The interference of HEPES buffer during amperometric detection of ATP in clinical applications

Jean Francois Masson; Estelle Gauda; Boris Mizaikoff; Christine Kranz

doi:10.1007/s00216-008-2015-y

The interference of HEPES buffer during amperometric detection of ATP in clinical applications

Jean Francois Masson, Estelle Gauda, Boris Mizaikoff, Christine Kranz

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

HEPES-based biological buffer is subject to photooxidation upon exposure to fluorescent illumination. Thereby hydrogen peroxide is generated, which interferes with amperometric oxidoreductase-based biosensors for glucose or adenosine triphosphate (ATP). These biosensors operate at an oxidation potential above 500 mV vs. the standard calomel electrode (SCE) and involve hydrogen peroxide as the electroactive molecule detected at the electrode surface. False-positive detection of ATP was observed in HEPES buffer utilizing an amperometric microbiosensor based on the co-immobilization of glucose oxidase and hexokinase for detection of ATP in biological specimens. Electrochemical, mass spectrometric, ³¹P NMR, and ¹H NMR studies indicate that complexation of ATP and HEPES induced by the presence of Ca²⁺ in HEPES buffer decreases the photooxidation of HEPES. Consequently, the hydrogen peroxide background concentration is reduced, thereby leading to erroneous ATP detection at the dual-enzyme microbiosensor, which determines an increase in ATP via a reduced hydrogen peroxide signal.

Original language	English (US)
Pages (from-to)	2067-2071
Number of pages	5
Journal	Analytical and Bioanalytical Chemistry
Volume	390
Issue number	8
DOIs	https://doi.org/10.1007/s00216-008-2015-y
State	Published - Apr 2008
Externally published	Yes

Keywords

4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES)
ATP microbiosensor
Glucose oxidase
Hexokinase
Phototoxicity
Ringer's solution

ASJC Scopus subject areas

Analytical Chemistry
Biochemistry

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title = "The interference of HEPES buffer during amperometric detection of ATP in clinical applications",

abstract = "HEPES-based biological buffer is subject to photooxidation upon exposure to fluorescent illumination. Thereby hydrogen peroxide is generated, which interferes with amperometric oxidoreductase-based biosensors for glucose or adenosine triphosphate (ATP). These biosensors operate at an oxidation potential above 500 mV vs. the standard calomel electrode (SCE) and involve hydrogen peroxide as the electroactive molecule detected at the electrode surface. False-positive detection of ATP was observed in HEPES buffer utilizing an amperometric microbiosensor based on the co-immobilization of glucose oxidase and hexokinase for detection of ATP in biological specimens. Electrochemical, mass spectrometric, 31P NMR, and 1H NMR studies indicate that complexation of ATP and HEPES induced by the presence of Ca2+ in HEPES buffer decreases the photooxidation of HEPES. Consequently, the hydrogen peroxide background concentration is reduced, thereby leading to erroneous ATP detection at the dual-enzyme microbiosensor, which determines an increase in ATP via a reduced hydrogen peroxide signal.",

keywords = "4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), ATP microbiosensor, Glucose oxidase, Hexokinase, Phototoxicity, Ringer's solution",

author = "Masson, {Jean Francois} and Estelle Gauda and Boris Mizaikoff and Christine Kranz",

note = "Funding Information: Acknowledgments The authors would like to thank Dr Leslie Gelbaum of the NMR Center, School of Chemistry and Biochemistry, Georgia Tech, for valuable help with acquisition and interpretation of NMR spectra. The authors acknowledge financial support by NIH grant R01 HL080725 and R21 HL082860.",

year = "2008",

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doi = "10.1007/s00216-008-2015-y",

language = "English (US)",

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T1 - The interference of HEPES buffer during amperometric detection of ATP in clinical applications

AU - Masson, Jean Francois

AU - Gauda, Estelle

AU - Mizaikoff, Boris

AU - Kranz, Christine

N1 - Funding Information: Acknowledgments The authors would like to thank Dr Leslie Gelbaum of the NMR Center, School of Chemistry and Biochemistry, Georgia Tech, for valuable help with acquisition and interpretation of NMR spectra. The authors acknowledge financial support by NIH grant R01 HL080725 and R21 HL082860.

PY - 2008/4

Y1 - 2008/4

N2 - HEPES-based biological buffer is subject to photooxidation upon exposure to fluorescent illumination. Thereby hydrogen peroxide is generated, which interferes with amperometric oxidoreductase-based biosensors for glucose or adenosine triphosphate (ATP). These biosensors operate at an oxidation potential above 500 mV vs. the standard calomel electrode (SCE) and involve hydrogen peroxide as the electroactive molecule detected at the electrode surface. False-positive detection of ATP was observed in HEPES buffer utilizing an amperometric microbiosensor based on the co-immobilization of glucose oxidase and hexokinase for detection of ATP in biological specimens. Electrochemical, mass spectrometric, 31P NMR, and 1H NMR studies indicate that complexation of ATP and HEPES induced by the presence of Ca2+ in HEPES buffer decreases the photooxidation of HEPES. Consequently, the hydrogen peroxide background concentration is reduced, thereby leading to erroneous ATP detection at the dual-enzyme microbiosensor, which determines an increase in ATP via a reduced hydrogen peroxide signal.

AB - HEPES-based biological buffer is subject to photooxidation upon exposure to fluorescent illumination. Thereby hydrogen peroxide is generated, which interferes with amperometric oxidoreductase-based biosensors for glucose or adenosine triphosphate (ATP). These biosensors operate at an oxidation potential above 500 mV vs. the standard calomel electrode (SCE) and involve hydrogen peroxide as the electroactive molecule detected at the electrode surface. False-positive detection of ATP was observed in HEPES buffer utilizing an amperometric microbiosensor based on the co-immobilization of glucose oxidase and hexokinase for detection of ATP in biological specimens. Electrochemical, mass spectrometric, 31P NMR, and 1H NMR studies indicate that complexation of ATP and HEPES induced by the presence of Ca2+ in HEPES buffer decreases the photooxidation of HEPES. Consequently, the hydrogen peroxide background concentration is reduced, thereby leading to erroneous ATP detection at the dual-enzyme microbiosensor, which determines an increase in ATP via a reduced hydrogen peroxide signal.

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KW - ATP microbiosensor

KW - Glucose oxidase

KW - Hexokinase

KW - Phototoxicity

KW - Ringer's solution

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The interference of HEPES buffer during amperometric detection of ATP in clinical applications

Abstract

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