METHOD AND REAGENT KIT FOR DETERMINING ACTIVITY OF 5'-NUCLEOTIDASE

摘要

The invention relates to a method for determining activity of 5'-nucleotidase, in which a biological sample is incubated in a manner and under conditions known per se with a nucleotide pentose monophosphate as substrate, the liberated inorganic phosphate is converted into a coloured complex by treating it with ammonium molybdate and a reducing agent, colour intensity is measured by a known method, and from the measured value the activity of 5'-nucle-otidase or the amount of inorganic phosphate liberated within unit time, as a figure proportional to the activity of 5'-nulceotidase, is calculated with a known calculation and/or with the aid of a calibration curve. According to the invention 5'-AMP, 5'-CMP, 5'-UMP, 5'-GMP, 5'-IMP and 5'-TMP are used as nucleotide pentose monophosphates, and measurement is performed on all of these six substrates. The invention also relates to a reagent kit for performing the above method.;Predmetni pronalazak se odnosi na postupak odredjivanja aktivnosti 5'-nukleotidaze, tokom koga se bioloki uzorak inkubira na nain i pod uslovima koji su uobiajeni za nukleotid pentozo-monofosfat kao supstrat, oslobodjeni neorganski fosfat se prevodi u obojeni kompleks reakcijom sa amonijum molibdatom kao redukcionim sredstvom, intenzitet obojenosti se meri poznatim postupkom, a na osnovu izmerene vrednosti aktivnosti 5'-nukleotidaze, ili na osnovu koliine neorganskog fosfata oslobodjenog u jedinici vremena, kao parametra koji je proporcionalan aktivnosti 5'-nukleotidaze, pomenuta aktivnost se izraunava poznatim postupkom za izraunavanje i/ili uz pomo kalibracione krive. Prema predmetnom pronalasku, 5'-AMP, 5'-CMP, 5'-UMP, 5'-GMP, 5'-IMP i 5'-TMP se koriste kao nukleotid pentozo-monofosfati, a merenja se vre na svim ovim supstratima. Predmetni pronalazak se takodje odnosi na komplet reagensa potrebnih za izvodjenje gore pomenutog postupka.[The invention relates to a method for determining activity of 5'-nucleotidase, in which a biological sample is incubated in a manner and under conditions known per se with a nucleotide pentose monophosphate as substrate, the liberated inorganic phosphate is converted into a coloured complex by treating it with ammonium molybdate and a reducing agent, colour intensity is measured by a known method, and from the measured value the activity of 5'-nucle-otidase or the amount of inorganic phosphate liberated within unit time, as a figure proportional to the activity of 5'-nulceotidase, is calculated with a known calculation and/or with the aid of a calibration curve. According to the invention 5'-AMP, 5'-CMP, 5'-UMP, 5'-GMP, 5'-IMP and 5'-TMP are used as nucleotide pentose monophosphates, and measurement is performed on all of these six substrates. The invention also relates to a reagent kit for performing the above method.