Suggested isosbestic wavelength calibration in clinical analyses

Abstract

The use of isosbestic points for conveniently checking the wavelength scale of spectrophotometers in the ultraviolet and visible regions is recommended. Colorimetric pH indicators, hemoglobin derivatives, and other radiation-absorbing substances that are convertible into stable isomers of different absorption spectra provide a means for calibrating many different wavelengths by comparing the absorptivities of these isomers in equimolar solutions. The method requires no special precautions and results are independent of substance concentration and temperature between 4 and 45°C. Isosbestic calibration may be important for (e.g.) coenzyme-dependent dehydrogenase activity determinations and in quality assurance programs.