Introduction Notwithstanding recent progress on molecular mechanisms underlying heart failure with preserved ejection fraction (HFpEF), multiple pathophysiological aspects of this condition including the basis of anaemia and other haematological disorders remain unresolved. In this study, we sought to determine the relationship of plasma volume (PV), a plausible confounding factor for the concentration of solutes in blood, with key haematological markers in HFpEF patients. Methods Total circulating PV was determined with high precision, automated carbon monoxide rebreathing in 24 stable HFpEF patients (70 AE 8 years, left ventricular ejection fraction = 55AE5%) and 18 healthy age-and sex-matched control (HC) individuals. Linear regression analyses were performed to determine the association of PV with a comprehensive set of haematological variables. Results Haematocrit (40Á1 AE 4Á9 versus 43Á6 AE 2Á7%, P = 0Á004) and haemoglobin concentration (131 AE 16 versus 142 AE 7 g l À1 , P = 0Á003) were reduced in HFpEF patients compared with HC individuals. In regression analyses, PV was negatively associated with haematocrit (r = À0Á45, P = 0Á029) and haemoglobin concentration (r = À0Á44, P = 0Á030) in HFpEF patients, whereas these variables were not associated with PV in HC individuals (P≥0Á198). Regarding blood electrolytes, PV was negatively associated with K + (r = À0Á43, P = 0Á036) and Ca 2+ (r = À0Á44, P = 0Á032) in HFpEF patients but not in HC individuals (P≥0Á734). None of the above associations were detected in HFpEF patients when using ideal instead of measured PV. Conclusion The blood concentration of routine markers of anaemia and electrolyte balance is specifically and linearly associated with PV in HFpEF patients. Excess or deficit of circulating PV may confound clinical diagnosis in this population.