Abstract Background The recently FDA-approved OrganOx Metra normothermic perfusion system for maintaining donor livers under physiologic conditions prior to transplantation comes equipped with a limited onboard blood gas analyzer for monitoring pO2, pCO2 and temperature. In clinical practice, however, additional biomarkers are used to assess the functional performance of the donor liver, including perfusate lactate clearance and glucose utilization. Liver perfusate contains a mixture of packed red blood cells, albumin, heparin, antibiotic, and solutions of amino acids and electrolytes and bile salts to support metabolic function, and is a specimen type that is not approved by the FDA. Implementation of the OrganOx Metra at Loyola University Medical Center (LUMC), a quaternary care transplant center, led to the validation of liver perfusate for the evaluation of glucose and lactate at the point-of-care (POC) using the Nova STATStrip Glucometer and Abbott iSTAT CG4 cartridge, respectively. Methods Lactate was measured on the Abbott iStat CG4 cartridge, and glucose which was measured using the Nova STATStrip Glucometer. Recovery studies were performed by mixing liver perfusate (non-centrifuged, n=16 independent specimen) with heparinized whole blood. Results The average recovery for the evaluation of lactate was 100% (range = 78 to 122%), and the average recovery for glucose was 109% (range = 93 to 112%). Conclusions Recovery studies evaluating matrix effect for the analysis of lactate (Abbott iSTAT, CG4) and glucose (Nova STATStrip) in liver perfusate indicate that the matrix of the perfusate does not affect the analytical accuracy, as compared to heparinized whole blood, the FDA-approved specimen type. Measurement of the OrganOx liver perfusate for lactate and glucose on these POC devices demonstrates reliable assessments to guide transplant teams in evaluating the viability of donor livers.