There are several point-of-care testing (POCT) devices for measuring critical laboratory values, but real-time utility can be cumbersome considering need for blood draws, multiple cartridges, and potential for delays in obtaining results. Digital Blood Corporation (DBC) developed an algorithm for non-invasive, real-time measurement of multiple values. The objective of this study was to compare the values obtained using a POCT device with the DBC’s non-invasive methodology and to evaluate the robustness of the algorithm. After Institutional Review Board approval, a pilot and feasibility study was conducted in healthy ambulatory individuals aged 18-64 years. Radial arterial blood was collected for the POCT comparator analysis using the Abbott i-STAT® device. The four parameters directly measured for DBC’s algorithm included temperature, hemoglobin, and partial pressures of oxygen and carbon dioxide (pO2 and pCO2, respectively). Using these values, the algorithm calculated sodium, potassium, chloride, ionized calcium (iCa), total carbon dioxide (TCO2), pH, bicarbonate, and oxygen saturation (SO2). Bland-Altman difference plot were calculated for analyzing the agreement between the Abbott i-STAT® device and DBC’s non-invasive methodology. For a second set of data, pO2 and pCO2 values collected using the Abbott i-STAT® device were used as input for DBC’s algorithm to test robustness of the algorithm. Data from 37 participants (mean age: 42.4 + 13 years; range: 18-64 years) were included in the primary analysis. Mean difference was less than 5% for sodium, chloride, pH, SO2, and bicarbonate; 6-10% for hemoglobin, TCO2 and pCO2; and greater than 10% for iCA, potassium, and pO2. The greatest variation between POCT and DBC’s approach was observed for pO2. Algorithm values that depend upon pO2 precision obtained from the TINA TCM4 radiometer are expected to show greatest deviation. Replacing transcutaneous pO2 and pCO2 values from the TINA TCM4 instrument with Abbott i-STAT values demonstrates the robustness of DBC’s algorithm and its ability to predict blood values comparable to the POC device in healthy patients. This pilot study serves as a proof of concept to trigger future study and further development of the DBC’s non-invasive device in critical care settings and test the usability of the device via quantitative and qualitative measures with different healthcare providers (e.g., pharmacists, nurses, paramedics). The broader clinical significance especially among critically ill adults and children remains to be determined.