Stability of glutathione in induced sputum: Impact of freezing

Background: Oxidative stress has repeatedly been linked to the pathogenesis of pulmonary disorders like asthma and chronic obstructive pulmonary disease. Measuring glutathione (GSH) in induced sputum (IS) offers a noninvasive tool to study oxidative stress in airway diseases. Objectives: This study assessed the stability of GSH in sputum supernatant under varying conditions. Methods: GSH in IS of 14 (7 healthy, 4 chronic obstructive pulmonary disease, 3 allergic rhinitis) nonsmoking subjects was quantified spectrophotometrically. The stability of GSH in supernatant was analyzed over 24 h under different ambient conditions (room temperature and cooling at 4degreesC). Reproducibility of GSH measurements in immediately processed and frozen supernatant (+72 h) was expressed by intraclass correlation coefficient (R-i) and coefficient of repeatability (CR). Results: GSH recovery in supernatant decreased in a time- and temperature-dependent manner. Samples stored at 4degreesC and room temperature showed a rapid decline of stability after 2 h. Mean GSH concentrations in IS after freezing (-20degreesC) and thawing after 72 h were not significantly different from GSH values measured immediately after processing of the samples ( immediate processing: 17.9 B 13.9 muM; 72 h freezing: 16.4 +/- 12.9 muM, p = 0.2). The reproducibility between immediately processed and frozen samples was excellent (R-i = 0.97; CR = 5.7 muM). Conclusions: Storage of sputum supernatant at room temperature or 4degreesC leads to a rapid decline of GSH recovery compared with baseline values. Immediate freezing of samples is a suitable and valid alternative to rapid processing and allows collection and shipment of samples for subsequent analysis. Copyright (C) 2003 S. Karger AG, Basel.