Method for improved accuracy in endogenous urea recovery marker calibrations for microdialysis in tumors.

Introduction: Urea has been proposed as an endogenous recovery marker for microdialysis for absolute concentration calculations of analytes in microdialysis samples. Previously we demonstrated a linear relationship between urea concentrations in a rat mammary carcinoma and that in plasma, validating its use as a recovery marker for that particular tumor. In this paper, we have extended the validation to two other tumor lines, thereby providing confidence that the calibration is constant across tumor types. To improve the accuracy in the determination of the plasma/tumor urea relationship from no net flux calibrations, we extended the range of the calibration by adding exogenous urea to tumor bearing animals. This method enabled more accurate calculations of absolute recovery from plasma and dialysate urea concentrations. We confirm that by using this method the calibration is valid across three different tumor lines. The existence of a common calibration between tumors provides rationale for using plasma urea as a recovery marker for clinical trials. The existence of a common calibration between tumor types bypasses the need to perform time consuming calibrations for each patient. This makes the procedure much more practical for clinical studies.

Methods: The no net flux technique was used to determine the plasma vs. tumor urea relationship for the R3230Ac mammary carcinoma, 9 L glioma, and a fibrosarcoma (FSa), grown in Fischer 344 rats. Plasma urea was stably increased beyond the normally occurring concentration for some of the data points by subcutaneous bolus administration to extend the range of data for the no net flux calibration.

Results: Urea recovery was unaffected by plasma urea concentration and was consistent with other reported values. The relationship between plasma and tumor urea was fit by a line, and linear regressions of the data with the extended plasma urea range had better R2 values than we reported previously. Statistical comparison of the regressions suggests that within reasonable uncertainty limits, they are the same for the different tumor types.

Discussion: Increasing the plasma urea concentration range for no net flux calibrations of urea as an endogenous recovery marker in tumors resulted in more accurate determination of the plasma/tumor urea relationship. A single linear regression may describe the relationship between plasma and tumor urea concentration across tumor lines for a given set of microdialysis parameters.