Journal article
Acridinium ester chemiluminescence: pH dependent hydrolysis of reagents and flow injection analysis of hydrogen peroxide and glutamate
Mikrochimica acta (1966), Vol.108(3-6), pp.205-219
05/1992
DOI: 10.1007/BF01242430
Abstract
An automated analysis system is described for the measurement of hydrogen peroxide based on a chemiluminescence reaction with phenyl 10-methylacridinium-9-carboxylate (PMAC). A reversed FIA experimental arrangement is used to establish the operating conditions for the measurement of submicromolar levels of hydrogen peroxide. The carrier stream consists of hydrogen peroxide standards prepared in a pH 9.0, boric acid buffer and the flow rate for this carrier/sample stream is 4 ml/min. Twenty microliters of a 10 mM PMAC solution, prepared in a pH 3 phosphate buffer, are injected into the carrier/sample stream. Hydrogen peroxide mixes with the PMAC reagent in an incubation coil that is constructed by wrapping 107 cm of polyethylene tubing around a 1 cm o.d. plastic rod. The chemiluminescence reaction is then initiated by adding base just before the sample passes in front of a photomultiplier tube (PMT) detector. The calculated limit of detection (S/N = 3) for hydrogen peroxide is 0.25 μM. In addition, the pH dependent hydrolysis of the PMAC reagent is characterized by an HPLC method which has been specifically developed for the separation and detection of the hydrolysis products of PMAC. Results indicate that a pH of 3.0 is required for long term stability of the PMAC reagent. Finally, this system has been successfully extended to the measurement of glutamate by coupling a bioreactor column of glutamate oxidase with the hydrogen peroxide detection scheme. A detection limit (S/N = 3) of 0.5 μM has been established for glutamate with a throughput of 200 samples per hour.
Details
- Title: Subtitle
- Acridinium ester chemiluminescence: pH dependent hydrolysis of reagents and flow injection analysis of hydrogen peroxide and glutamate
- Creators
- Maureen Stuever KaltenbachMark A Arnold
- Resource Type
- Journal article
- Publication Details
- Mikrochimica acta (1966), Vol.108(3-6), pp.205-219
- DOI
- 10.1007/BF01242430
- ISSN
- 0026-3672
- eISSN
- 1436-5073
- Language
- English
- Date published
- 05/1992
- Academic Unit
- Fraternal Order of Eagles Diabetes Research Center; Chemistry; Center for Biocatalysis and Bioprocessing
- Record Identifier
- 9984216561602771
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