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अमूर्त

Understanding Sample Preparation Workflow: Semi-Automated Preparation of Serum Testosterone for Detection Using LC-MS/MS

Ramisa Fariha, Collin Hill, Mohannad Jabrah, Adam Spooner, Anubhav Tripathi*

High-performance liquid chromatography-tandem mass spectrometry has been pivotal to recent clinical advancements in the detection and monitoring of the progression of different metabolic disorders. Such systems have been used to routinely monitor serum testosterone levels in patients, to both mitigate side effects of different medications, as well as diagnose androgen-associated disorders. The present study develops and validates an LC-MS/MS-based assay for the detection of total serum testosterone from only (100 μL) of sample volume. To increase the high throughput capability of the developed diagnostic kit for clinical use on patient samples, the proposed method was streamlined via semi-automation on a JANUS^® G3 Workstation liquid handler. The proposed diagnostic kit showcased high detection precision (CV<5%) as well as sample accuracy (93%-108%), within a clinically relevant range of testosterone serum concentrations (5-1500 ng/dl). Moreover, the assay showcased LOD of 5 ng/dl, which is significantly comparable to commercially established diagnostic tools. The developed method also exhibited high linearity over the specified testosterone serum range, with an obtained linearity of R2>0.99 using both manual as well as automated sample preparation. Overall, the work shown presents an optimized, automatable LC-MS/MSbased testosterone diagnostic assay for the routine evaluation of patient testosterone levels in serum for clinical and research use.