Tebuconazole and penconazole – Determination of TEB-OH, TEB-COOH, PEN-OH, and PEN-COOH in urine by LC-MS/MS

Biomonitoring Method – Translation of the German version from 2026

Elisa Polledri¹ (Method development)
Rosa Mercadante¹ (Method development)
Silvia Fustinoni¹ (Method development)
Laura Kuhlmann² (External verification)
  Thomas Göen² (External verification; Head of the working group “Analyses in Biological Materials” of the Permanent Senate Commission for the Investigation of Health Hazards of Chemical Compounds in the Work Area, Deutsche Forschungsgemeinschaft)
  Andrea Hartwig³ (Chair of the Permanent Senate Commission for the Investigation of Health Hazards of Chemical Compounds in the Work Area, Deutsche Forschungsgemeinschaft)
  MAK Commission<sup>4

1</sup> Laboratory of Environmental and Occupational Toxicology, Department of Clinical Sciences and Community Health, University of Milano and Fondazione IRCCS Ca Granda Ospedale Maggiore Policlinico, Via Francesco Sforza 35, 20122 Milano, Italy
² Friedrich-Alexander-Universität Erlangen-Nürnberg, Institute and Outpatient Clinic of Occupational, Social, and Environmental Medicine, Henkestraße 9–11, 91054 Erlangen, Germany
³ Institute of Applied Biosciences, Department of Food Chemistry and Toxicology, Karlsruhe Institute of Technology (KIT), Adenauerring 20a, Building 50.41, 76131 Karlsruhe, Germany
⁴ Permanent Senate Commission for the Investigation of Health Hazards of Chemical Compounds in the Work Area, Deutsche Forschungsgemeinschaft, Kennedyallee 40, 53175 Bonn, Germany

Abstract

The working group “Analyses in Biological Materials” of the German Senate Commission for the Investigation of Health Hazards of Chemical Compounds in the Work Area (MAK Commission) developed and verified this biomonitoring method for the measurement of the most important urinary metabolites of the triazole fungicides tebuconazole and penconazole. Specifically, this method determines (RS)-5‑(4‑chlorophenyl)-2,2‑dimethyl-3‑(1H‑1,2,4‑triazol-1‑ylmethyl)-1,3‑pentanediol (TEB‑OH) and (RS)-5‑(4‑chlorophenyl)-2,2‑dimethyl-3‑(1H‑1,2,4‑triazol-1‑ylmethyl)-3‑olpentanoic acid (TEB‑COOH) as well as 4‑(2,4‑dichlorophenyl)-5‑[1,2,4]‑triazol-1‑ylpentanol (PEN‑OH) and 4‑(2,4‑dichlorophenyl)-5‑[1,2,4]‑triazol-1‑ylpentanoic acid (PEN‑COOH) in urine. After adding isotope-labelled internal standards, the samples are enzymatically hydrolysed to release the analytes from the glucuronide and sulfate conjugates. After online purification, the analytes are separated by liquid chromatography and analysed using tandem mass spectrometry. Calibration is performed using calibration standards prepared in pooled urine and processed analogously to the samples to be analysed. The method provides reliable and accurate analytical results, as shown by the good precision data with standard deviations in the range of 0.1–10.7%. Good accuracy data were obtained with mean relative recoveries in the range of 100–107%. The method is selective, sensitive, and provides quantitation limits of 0.3 μg/l for TEB‑OH and TEB‑COOH and of 1.0 μg/l for PEN‑OH and PEN‑COOH.

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