Paraoxonase-1 (PON1) s an hgh densty lpoproten (HDL)-assocated enzyme wth antoxdant and antatherogenc functons, protectng lpoprotens aganst oxdatve modﬁcaton It also catalyzes the hydrolyss of organophosphates such as paraoxon and aromatc carboxylc acd esters of fatty acds. It has been shown that serum paraoxonase actvty decrease n dabetes melltus, coronary artery dsease, hypercholesterolaema, ron deﬁcency annema, heepatts, crrhoss, prostate cancer, tuberculoss and nﬂa mattons.
Fully automated paraoxonase actvty measurement method conssts of two dﬀerent seequenta reagents. The ﬁrst reagent s an approprate Trs buﬀer and t also contans calcum on, whch s a cofactor of PON1 enzyme. The second reagent s a new developed stable substrate soluton. The sample s mxed wth the Reagent 1 and the substrate soluton s added. Lnear ncrease of the absorbance of p-ntrophenol, produced from paraoxon, s followed at knetc measurement mode. Nonenzymatc hydrolyss of paraoxon was substracted from the total rate of hydrolyss. The molar absorptvty of p-ntrophenol s 18,290 M-1 cm-1 and one unt of paraoxonase actvty s equal to 1 mol of paraoxon hydrolyzed per lter per mnute at 37oC
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