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Ambient temperature may affect venous glucose concentration after glucose tolerance tests. We analysed 1030 standardised 75 g tests. Although mean fasting values did not differ, post-load values did: adjusted mean 2 h glucose concentration was 1·03 mmol/L lower at lower (5-14°C) than at higher (25-31°C) temperatures (p<0·001). The occurrence of abnormal glucose tolerance doubled on warmer days. The diagnostic accuracy of the glucose tolerance test showed clinically significant temperature-associated variation. These variations, if confirmed, call for temperature standardisation during glucose tolerance testing and/or alternative strategies for use when standardisation is not feasible.