Is It Required to Abstain From Fluid Consumption in the 10 Min Before Collection of a Saliva Sample?

The noninvasive and reliable saliva collection method for stress-related endocrine biomarkers analysis has become common in field- and laboratory-based research in the last 15 years. Current guidelines recommend interrupting water consumption 10 min before saliva sampling to avoid inaccurate, diluted concentrations of the biomarkers being examined. However, the impact of saliva dilution on salivary cortisol and testosterone levels has not been examined yet. This was a randomized, crossover experiment. Fourteen active healthy males completed three separate cycle-bout trials, where a set volume of water (250 ml) was consumed 10, 5, or 1 min before saliva sampling. Saliva was collected pre-, post-, and 30 min postexercise. No differences were observed in resting samples in any of the trials (p < .05). However, salivary cortisol increased from pre- to postexercise in the 10 (+52%, p = 0.022) and 5-min trial (+36%, p = .005) only. Salivary testosterone also increased in 10 (+34%, p = .005), 5 (+37%, p < .001), and 1-min trials (+24%, p < .035). This study proposes that consuming water up to 1 min before providing a saliva sample will not dilute the sample, allowing for an accurate determination of salivary cortisol and testosterone concentration levels. Practically, the 10-min fluid abstinence required before saliva sampling in the previous guidelines could be reduced to 1 min; however, if examining the response of salivary cortisol to an exercise trail, a fluid abstinence of 5 min may be more appropriate. This conclusion will allow for less restriction on researchers and participants alike.