TITLE:
A Rapid Screening Method for the Analysis of Beta-Blockers Using Electrospray Ionization-Liquid Chromatography-Tandem Mass Spectrometry
AUTHORS:
Keiko Tonooka-Kubota, Chizuko Sasaki, Masaru Terada, Fumiko Satoh, Tatsuo Shinozuka
KEYWORDS:
Beta Blocker, Analysis, LC-MS/MS, Intoxication, Solid-Phase Extraction
JOURNAL NAME:
American Journal of Analytical Chemistry,
Vol.17 No.4,
April
30,
2026
ABSTRACT: Cardiovascular drugs, when overdosed, may present serious symptoms of intoxication and can even be more severe than the underlying diseases themselves. The aim of this study was to develop and validate a sensitive electrospray ionization (ESI)-liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous analysis of 17 beta-blockers (Method A), beta-blockers (metoprolol, propranolol) and their metabolites (α-hydroxy metoprolol, O-desmethyl metoprolol, and 4-hydroxy propranolol) (Method B) in serum and urine. The extraction of beta-blockers and their metabolites from human serum and urine was performed by a solid-phase extraction method using Oasis® PRiME HLB columns. Chromatographic separation was performed on a Mightysil-RP-18 MS column. For gradient elution, two solvents were used: a) 10 mM acetic ammonium buffer and b) acetonitrile. Separation and sensitivity for the detection of beta-blockers and their metabolites by LC-MS/MS were sufficient, and the precursor [M + H]+ ion was detected in the mass spectrum of each drug (S/N = 3). These methods had a total chromatographic run time of 15 min. The calibration curves were linear over the concentration range of 20 - 400 ng/mL for the 17 beta-blockers (Method A) and 2 - 200 ng/mL (Method B). The extraction yields for human serum samples (17 beta-blockers) ranged from 66.1% - 93.5%, and the accuracy and precision were within 7.0% CV values, while the extraction yields for human serum or urine samples ranged from 93.8% to 106.5%, and the accuracy and precision were within 5.9% CV values, which were acceptable for these methods. The limits of quantification (LOQ) can cover the therapeutic blood concentration range in which beta-blockers are clinically used (Method A and Method B). The presently established method is useful for the simultaneous measurement of beta-blockers and their metabolites in human serum and urine by LC-MS/MS in clinical and forensic investigations.