Pleural fluid pH and glucose levels are both recommended in the workup of pleural effusions. Whether their levels correlate and predict each other or contribute independent knowledge is unclear. We aimed to investigate the pH/glucose relationship, assess their concordance and ascertain whether performing both tests provides additional information to performing either test alone. The pH and glucose measurements from 2,971 pleural fluid samples, from three centers in Spain, UK and Australia, were categorized into Cancer (n=1,045), Infection (n=544), Tuberculosis (n=249) and Others (n=1,133) groups. The relationship between pH and glucose values and their concordance at clinically relevant cutoffs (pH 7.2 and glucose 3.3 mmol/L) were assessed. The mean pH of the cohort was 7.38 (SD 0.22) and median glucose 5.99 (range, 0.00-29.36) mmol/L. A regression model of the relationship between glucose (log-transformed) and pH with a restricted cubic spline showed linear (P<0.01) and nonlinear effects (P<0.01). The relationship was strong with a narrow confidence interval but the prediction interval was wide. Most (91.9%) samples were concordant using pH and glucose levels at cutoffs of 7.20 and 3.30 mmol/L respectively. Using pH alone, without glucose, captured 95.0% of the infection-related effusions with either pH or glucose below cutoff and glucose alone identified 91.7%. Pleural fluid pH and glucose have a strong non-linear relationship but, in most situations, the level of one cannot accurately predict the other. Concordance rates were high and either test is sufficient in the majority of cases.