Novel optical method for measuring D-glucose 6-phosphate

Ashutosh Sharma; Nigel S. M. Quantrill

doi:10.1117/12.180780

21 July 1994 Novel optical method for measuring D-glucose 6-phosphate

Ashutosh Sharma, Nigel S. M. Quantrill

Proceedings Volume 2136, Biochemical Diagnostic Instrumentation; (1994) https://doi.org/10.1117/12.180780
Event: OE/LASE '94, 1994, Los Angeles, CA, United States

Abstract

A novel D-glucose 6-phosphate (G6P) assay is described that exploits the technique of substrate induced quenching. In a reaction between G6P and glucose 6-phosphate dehydrogenase (G6P-DH) the cofactor nicotinamide adenine dinucleotide (NAD⁺) is reduced to NADH. The latter being a quencher of thionine allows an indirect determination of the G6P concentration. A Stern-Volmer type analysis of the data for the investigated range of 0 - 1 mM G6P results in a quenching constant of 439 (+/- 37) M^-1, with a detection limit of 9 (mu) M. A simple empirical model based on enzyme kinetics is presented for the overall G6P assay. Excellent correlation exists between the results predicted by the model and those obtained experimentally. Possibilities of employing the proposed G6P SIQ assay in multi-enzyme assays and in the construction of immunoassays is discussed.

Citation Download Citation

Ashutosh Sharma and Nigel S. M. Quantrill "Novel optical method for measuring D-glucose 6-phosphate", Proc. SPIE 2136, Biochemical Diagnostic Instrumentation, (21 July 1994); https://doi.org/10.1117/12.180780

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