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Saliva cotinine is commonly used to estimate nicotine intake but laboratories use different methods of collection. In three small trials, comparisons were made between (1) sugar vs. unstimulated saliva production (n= 29), (2) wax chewing vs. unstimulated production (n= 15) and (3) between two consecutive unstimulated saliva samples (n = 10). Sugar‐stimulated saliva cotinine scores were 26% below unstimulated levels (p < 0.001); correlation between measures was high (r = 0.90; p < 0.001), Wax stimulated saliva yielded levels 6% below unstimulated (p < 0.05; correlation: r = 0.98; p < 0.001). No differences were observed between two unstimulated samples taken within a ∼ 20‐minute period (correlation: r = 0.99; p < 0.001). It is postulated that changes in salivary flow can account for the findings.