Development of a technical colorimetric for the determination of galactosis in blood

Abstract

Galactosemia, a life-threatening disorder, is an inherited metabolic disease transmitted in an autosomal recessive manner. It is metabolized in humans and other species predominantly by the sequential activities of three enzymes that constitute the Leloir pathway. A deficiency of any one of the Leloir enzymes in humans, results in a galactosemia. Our study aims to develop a fast and accurate colorimetric technique of qualitative and quantitative determination of galactose in blood. This technique involves incubating the standard galactose and glucose ranges with orcinol in a strongly acidic solution to form a product capable of absorbing light at both wavelengths. This allows the determination of both sugars simultaneously. The standard range of 12.5 to 200 mg/linearity of y = 0.0027x - 0.004, R² = 0.9999 and y = 0.0017x + 0.0129, R² = 0.9963 of glucose and galactose respectively, agitation of orcinol is 3 min, incubation in the dark is 30min. The maximum absorbance of galactose is 569.5 nm while that of glucose is 421 nm. Since galactose in whole blood is unstable over time, all the parameters of the assay have been standardized after 1 year of testing. We have modied a technique colorimetrique that can be applied to the detection of galactosemia. The target analyte is total galactose. We continued to simplify this technique which is fast and economical. The interference between glucose and galactose has been bypassed by a simple calculation which makes it possible to determine the concertration of these two sugars simultaneously. To have good results, certain factors must be maintained, such as time and temperature. Thanks to this method we could diagnose 8 suspected cases of which only one has this disease.