Automated lab-on-valve sequential injection ELISA for determination of carbamazepine

Anal Chim Acta. 2019 Oct 17:1076:91-99. doi: 10.1016/j.aca.2019.05.017. Epub 2019 May 9.

Abstract

The development of an automated miniaturized analytical system that allows for the rapid monitoring of carbamazepine (CBZ) levels in serum and wastewater is proposed. Molecular recognition of CBZ was achieved through its selective interaction with microbeads carrying anti-CBZ antibodies. The proposed method combines the advantages of the micro-bead injection spectroscopy and of the flow-based platform lab-on-valve for implementation of automatic immunosorbent renewal, rendering a new recognition surface for each sample. The sequential (or simultaneous) perfusion of CBZ and the horseradish peroxidase-labelled CBZ through the microbeads is followed by real-time on-column monitoring of substrate (3,3',5,5'-tetramethylbenzidine) oxidation by colorimetry. The evaluation of the initial oxidation rate and also the absorbance value at a fixed time point provided a linear response versus the logarithm of the CBZ concentration. Under the selected assay conditions, a single analysis was completed after only 11 min, with a quantification range between 1.0 and 50 μg L-1. Detection of CBZ levels in undiluted wastewater samples was feasible after a simple filtration step while good recoveries were attained for spiked certified human serum, analyzed without sample clean-up.

Keywords: Automation; Bead injection spectroscopy; Human serum; Microparticles; Therapeutic drug monitoring; Wastewater.

MeSH terms

  • Antibodies, Monoclonal / immunology
  • Armoracia / enzymology
  • Benzidines / chemistry
  • Carbamazepine / blood*
  • Carbamazepine / immunology
  • Colorimetry / methods*
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Horseradish Peroxidase / chemistry
  • Humans
  • Microspheres
  • Oxidation-Reduction
  • Sepharose / analogs & derivatives
  • Sepharose / chemistry
  • Wastewater / analysis

Substances

  • Antibodies, Monoclonal
  • Benzidines
  • Waste Water
  • Carbamazepine
  • 3,3',5,5'-tetramethylbenzidine
  • Sepharose CL 4B
  • Sepharose
  • Horseradish Peroxidase