Automated Sample Preparation of Biological Fluids for LC-MS/MS Analysis

; ; (). Automated Sample Preparation of Biological Fluids for LC-MS/MS Analysis: Poster. In: MSACL 2014 US. 6th Annual Conference & Exhibits. (March 1-5, 2014). San Diego, CA: Association for Mass Spectrometry: Applications to the Clinical Lab (MSACL).

In laboratories using LC-MS/MS for analysis, labour intensive and time-consuming steps are increasingly being transferred to automated platforms. In order to properly automate these process steps, the operations must be simple and meet stringent criteria for robustness. All operations should be fully contained within the automation work table. Further, it is highly desirable to be able to perform automated method development on the same worktable. To address these needs a typical workflow for method development and sample preparation of biological fluids was automated using the AC Extraction Plate™.

This novel simplified extraction technology of small molecules from biological fluids has been adapted to a 96 wellplate format. The individual wells contain an innovative coating into which analytes absorb. Subsequent steps include washing each well followed by elution with the appropriate HPLC solvent mix. The three step approach reduces the number, as well as complexity, of the process steps to simple pipetting and shaking operations. The workflow is simple and hence perfectly suited for automation. This simple work flow has been adapted to robotic processing for sample preparation using a Tecan Freedom EVO. Scripts were developed for the liquid handling system taking advantage of the simple operation of AC Extraction Plate™.

  • The simple equilibrium process and protein removal process will be described.
  • A flow chart of the work flow for method development and sample processing using liquid handling automation will be explained.
  • The automated clean-up of the metabolites 25OH-Vitamin D3/D2 from human serum will be described including analytical linearity, reproducibility and robustness.
  • Additional analytes which have been successfully extracted using this workflow for method development including steroids from serum and THC metabolites from urine will be presented.

In summary, a general work table layout and robotic process for the 96 well format will be presented capable of automating both the method development and the sample processing of biological fluids for subsequent analysis by LC-MS/MS.