Method transfer and troubleshooting of testosterone, androstenedione and pregnenolone in human serum using liquid chromatography and tandem mass spectrometric detection

ASMS 2025 -- Biomarker analysis is critical for several reasons, especially in healthcare, research, and drug development. Biomarkers help detect diseases at an early stage, monitor disease progression, aid in tailored treatments to an individual’s genetic makeup, can be used to evaluate the safety and effectiveness of new drugs in clinical trials, and help detect harmful effects of drugs or environmental toxins before severe damage occurs. Method transfers from one laboratory to another often lead to unforeseen issues. During the method transfer to analyze testosterone, 4-androstene-3,17-dione and pregnenolone in human serum via Waters ultra performance liquid chromatography (UPLC) and mass spectrometery (MS) from one global laboratory to another, the following issues were observed and resolved at the transfer site: nonspecific binding (NSB) of the compounds, supported liquid extraction (SLE) plate lot variance, and baseline noise reduction.