Gómez E, Cladera A, Estela J M, Cerda V
Departament de Química, Universitat de les Illes Balears, 07071 Palma de Mallorca, Spain.
Talanta. 1993 Nov;40(11):1601-7. doi: 10.1016/0039-9140(93)80073-z.
This paper reports a multicomponent least-squares regression method based on the use of multiple standards for the simultaneous resolution of substrates involved in kinetic enzymatic processes. The proposed method was applied to the determination of catechol-resorcinol mixtures by oxidation with hydrogen peroxide in the presence of the enzyme peroxidase (EC 1. 11.1.7) in a stopped-flow reversed flow injection system. The mathematical algorithm used is superior to other alternatives as it is not affected by side reactions between the oxidation products. The proposed method allows the simultaneous resolution of 50-150,muM catechol and 30-180,muM resorcinol over the wavelength range 340-500 nm. The concentration ranges can be modified by varying the injected amount of enzyme. The mathematical treatment is applied to the spectra of several standards and those of the samples, which are recorded 6 sec after the flow is stopped. This redounds to a high sampling frequency (up to 60/hr).
