Reactive oxygenmetabolites measurement, establishment of a new method and its performance evaluation

Abstract

Objective To establish a simple, cost–effective method to measure reactive oxygen metabolites (ROMs) using automatic biochemical analyzer. Methods Serum samples were collected from 50 healthy individuals and 50 hospitalized patientsin Beijing Hospital from September 2013 to November 2013. By setting up and optimizing parameters on automatic biochemical analyzer, a new method of measuring ROMS was established with TBHP as calibrator.130 healthy non–smokers were recruited in Beijing Hospital from November 2013 to December 2013 to establish reference interval, including 73 males and 57 females. The average age was 68.8±12.5. According to CLSI documents, there was the establishment of the new method including precision, limit of blank (LoB), limit of detection (LoD), limit of quantitation (LoQ), linearity, interference testing, and reference interval. Results The intra assay imprecision was 1.5%–4.1% and the total imprecision was 4.4%–9.9%. LoB was 0.85 mmol/L TBHP; LoD was 2.7mmol/L TBHP; LoQ was 5.9mmol/L TBHP. The linearity was 5.9–600 mmol/L (R2=0.995). When triglyceride ≤28.58 mmol/L, hemoglobin ≤173 g/L, vitamin C ≤60 mg/dl and bilirubin≤ 73.8 μmol/L, the deviation was –7.6%, 4.6%, –98.9%, 19.1% respectively. The normal reference interval was 61.9–88.1 mmol/L TBHP. Conclusions The newly–established method has good performances of precision, LOD and linearity, which can meet the clinical needs. The interference of triglyceride and hemoglobin is small, while vitamin C and bilirubin have stronger interference on measurement. (Chin J Lab Med, 2015, 38: 163–167) Key words: Oxidative stress; Reactive oxygen species; Reference values