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Stable-labeled analogues and reliable quantification of nonprotein biomarkers by LC-MS/MS.

BACKGROUND
The aim was to develop, and establish as suitable to begin assessment by full validation, a quantitative LC-MS/MS method for asparagine in human plasma. Therein, to utilize a stable-labeled analogue of asparagine to act as surrogate analyte, producing complete calibration curves and corresponding QC samples and another m/z distinct stable-labeled analogue to act as internal standard.


RESULTS
From two candidates, the surrogate analyte was selected through statistical comparisons of concentration-response data and the resultant method employed protein precipitation and LC on an unmodified silica column with multiple reaction monitoring detection mode. The calibration range was 50-10,000 ng/ml.


CONCLUSION
This method was successfully proven to meet the accuracy and precision acceptance criteria of current bioanalytical method validation guidelines.