Quantification of endogenous steroids in human urine by gas chromatography mass spectrometry using a surrogate analyte approach

Providing “real blank sample” is a problem in determination of endogenous steroids in complex matrices. A new quantification strategy is proposed in the present study, which is based on using isotope-labeled steroids instead of natural steroids for constructing calibration line. This approach is called surrogate analyte and it is shown that its accuracy is better than some of the previously described methods at low concentrations and comparable to standard addition method at medium and high concentration levels. The method was fully validated to satisfy the ICH criteria and it was applied for determination of endogenous steroids in several urine samples.