ENHANCED CHEMILUMINESCENT ASSAY FOR MEASURING THE TOTAL ANTIOXIDANT CAPACITY OF SERUM, SALIVA AND CREVICULAR FLUID

This paper reports the development of an enhanced chemiluminescent (EC L) assay for measuring the total antioxidant (AO) capacity of serum, s aliva and a fluid collectable from the gum margin called gingival crev icular fluid (GCF), The theory behind the assay is explained, and the optimum conditions for the assay, and for storage of reagents and clin ical samples is described. Calibration lines were linear (R greater th an or equal to 0.99; P<0.0001) and the within batch coefficient of var iations for a water soluble vitamin E analogue (Trolox), serum and sal iva samples were <5%. In saliva and GCF, a characteristic AO response not seen in serum of the same patients, was identified. Total peripher al (serum) and local (saliva) AO capacities (mu mol/L Trolox) were inv estigated in patients with (n = 18) and without (n = 16) adult periodo ntitis. Serum AO status did not differ between groups. Salivary total AO concentrations were lower in the periodontitis (P) group [175 (53) mu mol/L] than in the non-periodontitis (NP) group [254 (110) mu mol/L -1: P<0.01], as were saliva:serum AO ratio's [0.37 (0.11) versus 0.5 ( 0.18): P<0.01]. Periodontitis patients may have a reduced salivary AO concentration, which could result from, or predispose to, the damaging effects of reactive oxygen species (ROS). The potential for ROS produ ction in the oral and periodontal environment may explain the presence of a specific antioxidant in oral fluids that is not detectable in se rum. The ECL assay described provides a rapid, simple and reproducible method of measuring total antioxidant defence in small volumes of bio logical fluids.