FIELD: chemistry.
SUBSTANCE: invention relates to biosensor technology, analytical chemistry and electrochemical determination of N-acetyl--β-D-glucosaminidase in biological fluids via amperometric determination of phenol which is released during enzymatic hydrolysis of 1-phenyl-M-acetyl-β-D-glucosaminide in biological fluids. The method involves adding to a biological fluid sample, containing N-acetyl-β-D-glucosaminidase, a solution of 1-phenyl-M-acetyl-β-D-glucosaminide in a nitrate buffer, followed by incubation, separation of the formed residue and obtaining a phenol-containing sample which is placed into electrochemical cell containing a buffer solution and an electrochemical tyrosinase biosensor, which is a planar graphite electrode obtained by screen printing, with successively deposited alternating adhesive layer(s) of polydiallyl dimethylammonium chloride and tyrosinase layer(s) as the working electrode, and an amperometric signal is detected, from which activity of N-acetyl-β-D-glucosaminidase is calculated based on a calibrated curve using a given equation.
EFFECT: high sensitivity and simple analysis.
2 cl, 5 ex, 4 dwg
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Authors
Dates
2012-11-20—Published
2011-06-10—Filed