Development of an automated method for the determination of human paraoxonase1 activity

Abstract

AbstractBackground: Human plasma paraoxonase1 (PON1) is an esterase catalyzing the hydrolysis of organophosphorus pesticides and other xenobiotics. The aims of this study were to develop a rapid method to determinate PON1 activity, evaluate some interference, and study the influence of storage temperature on PON1 activity assay.Methods: Measurement of PON1 activity was performed for 369 samples by measuring the hydrolysis of paraoxon using a spectrophotometric method adapted on konelab 30 ⃞.Results: The developed method facilitates the determination of PON1 activity at the rate of more than 200 samples per hour, and it is linear between 2 and 900 IU/L. Intra and inter-assay imprecision coefficients of variation were 2% and 5% respectively. PON1 activity in serum was correlated with those in heparinized plasma (r = 0.994, p < 0.001) and in plasma/EDTA (r = 0.962, p < 0.001). The mean inhibition of the PON1 activity was, by EDTA/K3, 41 ± 10 %. There was not significant PON1 activity variation after 40 days of storage at -20°C or at +4 ⃞ C. There were no substantial interferences from haemoglobin, jaundice and hyperlipidemia.Conclusion: The developed method is reliable, reproducible, and suitable. It can also be performed on heparinized plasma for the determination of PON1 activity. Hence, it may be useful for assaying PON1 activity in several intoxications such as organophosphorus, sarin, and soman nerve agents.