Solid-phase extraction and derivatisation methods for beta-blockers in human post mortem whole blood, urine and equine urine.

The research article discusses the development and testing of sensitive methodologies for the extraction and modification of certain beta-blockers in horse urine and human biological samples.

Objective of the Research

• The main goal of this study was to devise efficient and sensitive methods for the extraction and derivatisation of nine specific beta-blockers in horse urine and in human post mortem whole blood and urine. These drugs include propranolol, metoprolol, sotalol, atenolol, pindolol, timolol, oxprenolol, alprenolol, and penbutolol.

Methodology Employed

• The researchers employed three different solid-phase extraction techniques involving the use of XtrackT XRDAH515, Bond Elut Certify, or Sep-Pak C18 cartridges.
• Two derivatisation methods are also detailed in the study. These processes involve the creation of cyclised silyl or pentafluoropropionate derivatives with either chloromethyldimethylchlorosilane or pentafluoropropionic anhydride.
• The analysis of the samples was done through gas chromatographic-mass spectrometry in select-ion monitoring mode. This technique helps in the identification and quantification of the chemicals in the samples.

Evaluation and Application

• These developed methods were evaluated using samples from drug-free human post mortem blood, urine, and horse urine which were fortified at various levels with the above-mentioned beta-blockers.
• The researchers presented the application of some of these methods in a practical forensic case study to demonstrate their effectiveness and potential use in real-life scenarios involving forensic or anti-doping investigations.

Constraints of the Study

• Despite having wide-ranging implications, the study did not involve samples from equine administration trials of beta-blockers, limiting the scope of the experiment to only post mortem human samples and drug-free equine urine samples.