Skip to main content
SpringerLink
Log in

Determination of Thyreostats in Bovine Urine and Thyroid Glands by HPLC–MS/MS

  • Original
  • Published:
Chromatographia Aims and scope Submit manuscript

Abstract

The use of thyreostats in livestock is strictly forbidden by European legislation since 1981. The investigation of thyreostats is commonly performed by their detection as derivatives with 3-iodobenzylbromide. Although it has advantages, the derivatisation procedure can generally cause a decrease in analyte concentrations. With the aim of simplifying the analysis of five thyreostats in both bovine urine and in thyroid glands, two methods were developed without the derivatisation step. Salting-out assisted liquid–liquid extraction was carried out for both matrices, followed by high-performance liquid chromatography coupled with triple-quadrupole mass spectrometry analysis. The methods were validated in agreement with the guidelines of Commission Decision 2002/657/EC. For all the thyreostats evaluated, satisfactory results were achieved; the recovery was within 96–104 % for both the matrices, while precision (coefficient of variation) was less than 20 % for urine and 21 % for thyroid glands. The limits of decision and capacities of detection for all the compounds were lower than the recommended values of 10 μg L−1 and 10 μg kg−1, respectively. In urine, the limits of decision ranged from 6.9 to 7.3 μg L−1, and the capacities of detection ranged from 8.5 to 9.7 μg L−1, while in thyroid glands these values varied from 6.6 μg kg−1 to 7.4 μg kg−1 and from 8.0 μg g−1 to 9.7 μg kg−1, respectively. The results obtained show that the methods described are suitable for the direct detection of thyreostats in bovine urine and thyroid glands.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Palliola E, Moretti G, Nanni A, Amici M, Rossi C (1987) Valutazione degli esami istologico, ponderale e chimico sulle tiroidi dei bovini quali elementi per il controllo del trattamento illegale con tireostatici. Ann Ist Super Sanità 23:129–134

    CAS  Google Scholar 

  2. Pérez-Fernández V, Marchese S, Gentili A, García MÁ, Curini R, Caretti F, Perret D (2014) Analysis of antithyroid drugs in surface water by using liquid chromatography–tandem mass spectrometry. J Chromatogr A 1367:78–89. doi:10.1016/j.chroma.2014.09.045

    Article  Google Scholar 

  3. Hall JE, John E (2010) Guyton and Hall textbook of medical physiology. Elsevier Health Sciences, Philadelphia

    Google Scholar 

  4. De Wasch K, Brabander HB, Impens S, Vandewiele M, Courtheyn D (2001) Determination of mercaptobenzimidazol and other thyreostat residues in thyroid tissue and meat using high-performance liquid chromatography–mass spectrometry. J Chromatogr A 912:311–317. doi:10.1016/S0021-9673(01)00563-5

    Article  Google Scholar 

  5. Pinel G, Bichon E, Pouponneau K, Maume D, André F, Le Bizec B (2005) Multi-residue method for the determination of thyreostats in urine samples using liquid chromatography coupled to tandem mass spectrometry after derivatisation with 3-iodobenzylbromide. J Chromatogr A 1085:247–252. doi:10.1016/j.chroma.2005.06.055

    Article  CAS  Google Scholar 

  6. Vanden Bussche J, Noppe H, Verheyden K, Wille K, Pinel G, Le Bizec B, De Brabander HF (2009) Analysis of thyreostats: a history of 35 years. Anal Chim Acta 637:2–12. doi:10.1016/j.aca.2008.08.027

    Article  CAS  Google Scholar 

  7. Martinez-Frias ML, Cereijo A, Rodriguez-Pinilla E, Urioste M (1992) Methimazole in animal feed and congenital aplasia cutis. Lancet 339:742–743

    Article  CAS  Google Scholar 

  8. Eghbalian F (2007) Aplasia custis congenita after methimazole exposure in utero; a case report and literature review. Iran J Pediatr 17:293–296

    Google Scholar 

  9. Pinel G, Maume D, Deceuninck Y, Andre F, Le Bizec B (2006) Unambiguous identification of thiouracil residue in urine collected in non-treated bovine by tandem and high-resolution mass spectrometry. Rapid Commun Mass Spectrom 20:3183–3187. doi:10.1002/rcm.2711

    Article  CAS  Google Scholar 

  10. Brabander HB, Noppe H, Verheyden K, Vanden Bussche J, Wille K, Okerman L, Vanhaecke L, Reybroeck W, Ooghe S, Croubels S (2009) Residue analysis: future trends from a historical perspective. J Chromatogr A 1216:7964–7976. doi:10.1016/j.chroma.2009.02.027

    Article  Google Scholar 

  11. Vanden Bussche J, Vanhaecke L, Deceuninck Y, Verheyden K, Wille K, Bekaert K, Le Bizec B, De Brabander HF (2010) Development and validation of an ultra-high performance liquid chromatography tandem mass spectrometry method for quantifying thyreostats in urine without derivatisation. J Chromatogr A 1217:4285–4293. doi:10.1016/j.chroma.2010.04.030

    Article  CAS  Google Scholar 

  12. EC (1981) Council Directive 81/602/EC concerning the prohibition of certain substances having a hormonal action and of any substances having a thyrostatic action, Off J L 222, 31.07.1981, p. 32–33

  13. EC (1996) European Community Council Directive 96/23/EC of 29 April 1996 on measures to monitor certain substances and residues thereof in live animals and animal products and repealing Directives 85/358/EEC and 86/469/EEC and Decisions 89/187/EEC and 91/664/EEC, Off J L 125, 23.05.1996, pp. 10–32

  14. Vanden Bussche J, Kiebooms JAL, De Clercq N, Deceuninck Y, Le Bizec B, De Brabander HF, Vanhaecke L (2011) Feed or food responsible for the presence of low-level thiouracil in urine of livestock and humans? J Agr Food Chem 59:5786–5792. doi:10.1021/jf200556x

    Article  CAS  Google Scholar 

  15. Kiebooms JAL, Wauters J, Vanden Bussche J, Houf K, De Vos P, Van Trappen SI, Cleenwerck Vanhaecke L (2014) Identification and characterisation of thiouracil forming bacteria upon porcine in vitro digestion of Brassicaceae feed. Appl Environ Microbiol 80:7433–7442. doi:10.1128/AEM.02370-14

    Article  Google Scholar 

  16. Kiebooms JAL, Wauters J, Vanden Bussche J, Vanhaecke L (2015) Validation of a quantitative method using liquid chromatography coup led to multiple mass spectrometry for thiouracil in feedstuffs used in animal husbandry. Anal Bioanal Chem 407:4373–4384. doi:10.1007/s00216-014-8347-x

    Article  CAS  Google Scholar 

  17. EURL (2007) CRL guidance paper (7 December 2007): CRL’s view on state of art analytical methods for national residue control plans. http://www.bvl.bund.de/SharedDocs/Downloads/09_Untersuchungen/EURL_Empfehlungen_Konzentrationsauswahl_Methodenvalierungen_EN.pdf?__blob=publicationFile&v=2. Accessed 16 Mar 2016

  18. Wauters J, Vanden Bussche J, Le Bizec B, Kiebooms JAL, Dervilly-Pinel G, Prevost S, Wozniak B, Sterk SS, Grønningen D, Kennedy DG, Russell S, Delahaut P, Vanhaecke L (2015) Towards a new European threshold to discriminate illegally administered from naturally occurring thiouracil in livestock. J Agr Food Chem 63:1339–1346. doi:10.1021/jf504475f

    Article  CAS  Google Scholar 

  19. Ministero della Salute, Direzione Generale della Sicurezza degli Alimenti e della Nutrizione, relazione Finale, Piano Nazionale Residui 2015. http://www.izslt.it/new/wp-content/uploads/2015/06/PNR-2015. Accessed 16 Mar 2016

  20. Lõhmus M, Kallaste K, Le Bizec B (2009) Determination of thyreostats in urine and thyroid gland by ultra-high performance liquid chromatography tandem mass spectrometry. J Chromatogr A 1216:8080–8089. doi:10.1016/j.chroma.2009.04.005

    Article  Google Scholar 

  21. Blanchflower WJ, Hughes PJ, Cannavan A, McCoy MA, Kennedy DG (1997) Determination of thyreostats in thyroid and urine using high-performance liquid chromatography–atmospheric pressure chemical ionisation mass spectrometry. Analyst 122:967–972. doi:10.1039/A700829E

    Article  CAS  Google Scholar 

  22. Abuín S, Centrich F, Rúbies A, Companyó R, Prat MD (2008) Analysis of thyreostatic drugs in thyroid samples by ultra-performance liquid chromatography tandem mass spectrometry detection. Anal Chim Acta 617:184–191. doi:10.1016/j.aca.2007.12.018

    Article  Google Scholar 

  23. Poole C (2009) Handbook of methods and instrumentation in separation science, vol 1. Academic Press, San Diego

    Google Scholar 

  24. EU (2002) Commission decision 2002/657/EC of 12 August 2002, concerning the performance of analytical methods and the interpretation of the results. Off J Eur Commun 2002:L221–L232

    Google Scholar 

  25. SANCO (2008) European Union, European Commission, Health & Consumer Protection, Directorate General Directorate E, Safety of the Food Chain, Document SANCO/2004/2726-revision 4, December 2008, Guidelines for the Implementation of Decision 2002/657/EC

  26. Wu H, Zhang J, Norem K, El-Shourbagy TA (2008) Simultaneous determination of a hydrophobic drug candidate and its metabolite in human plasma with salting-out assisted liquid/liquid extraction using a mass spectrometry friendly salt. J Pharmaceut Biomed 48:1243–1248. doi:10.1016/j.jpba.2008.09.002

    Article  CAS  Google Scholar 

  27. Zhang J, Wu H, Kim E, El-Shourbagy TA (2009) Salting-out assisted liquid/liquid extraction with acetonitrile: a new high throughput sample preparation technique for good laboratory practice bioanalysis using liquid chromatography–mass spectrometry. Biomed Chromatogr 23:419–425. doi:10.1002/bmc.1135

    Article  CAS  Google Scholar 

  28. Song S, Ediage EN, Wu A, De Saeger S (2013) Development and application of salting-out assisted liquid/liquid extraction for multi-mycotoxin biomarkers analysis in pig urine with high performance liquid chromatography/tandem mass spectrometry. J Chromatogr A 1292:111–120. doi:10.1016/j.chroma.2012.10.071

    Article  CAS  Google Scholar 

  29. Thompson M (2000) Recent trends in inter-laboratory precision at ppb and sub-ppb concentrations in relation to fitness for purpose criteria in proficiency testing. Analyst 125:385–386. doi:10.1039/b000282h

    Article  CAS  Google Scholar 

  30. Abuín S, Companyó R, Centrich F, Rúbies A, Prat MD (2008) Analysis of thyreostatic drugs in thyroid samples by liquid chromatography tandem mass spectrometry: comparison of two sample treatment strategies. J Chromatogr A 1207:17–23. doi:10.1016/j.chroma.2008.08.018

    Article  Google Scholar 

  31. Galarini R, Piersanti A, Falasca S, Salamida S, Fioroni L (2007) A confirmatory method for detection of a banned substance: the validation experience of a routine EU laboratory. Anal Chim Acta 586:130–136. doi:10.1016/j.aca.2006.10.041

    Article  CAS  Google Scholar 

  32. Van Loco J, Jànosi A, Impens S, Fraselle S, Cornet V, Degroodt JM (2007) Calculation of the decision limit (CCα) and the detection capability (CCβ) for banned substances: the imperfect marriage between the quantitative and the qualitative criteria. Anal Chim Acta 586:8–12. doi:10.1016/j.aca.2006.11.058

    Article  Google Scholar 

  33. Gowik P (2009) The validation of methods for regulatory purposes in the control of residues. J Chromatogr A 1216:8051–8058. doi:10.1016/j.chroma.2009.06.059

    Article  CAS  Google Scholar 

  34. Lega F, Contiero L, Biancotto G, Angeletti R (2013) Determination of thyreostats in muscle and thyroid tissues by QuEChERS extraction and ultra-performance liquid chromatography tandem mass spectrometry. Food Addit Contam: Part A 30:949–957. doi:10.1080/19440049.2013.780212

    Article  CAS  Google Scholar 

Download references

Acknowledgments

Giuseppe Federico Labella is the recipient of a Ph.D. fellowship in Veterinary and Animal Science in the Laboratory of Veterinary Toxicology at the University of Milan. Elisa Pasquale is the recipient of a Cariplo Ph.D. fellowship in Animal Production in the Laboratory of Inspection of Food of Animal Origin at the University of Milan.

Author information

Authors and Affiliations

  1. Department of Veterinary Science and Public Health, University of Milan, Milan, Italy

    Giuseppe Federico Labella & Francesco Arioli

  2. Department of Health, Animal Science and Food Safety, University of Milan, Milan, Italy

    Luca Maria Chiesa, Elisa Pasquale, Sara Panseri & Radmila Pavlovic

Authors
  1. Luca Maria Chiesa
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Giuseppe Federico Labella
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Elisa Pasquale
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sara Panseri
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Radmila Pavlovic
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Francesco Arioli
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sara Panseri.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with animals performed by any of the authors.

Informed consent

Not applicable.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Chiesa, L.M., Labella, G.F., Pasquale, E. et al. Determination of Thyreostats in Bovine Urine and Thyroid Glands by HPLC–MS/MS. Chromatographia 79, 591–599 (2016). https://doi.org/10.1007/s10337-016-3068-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10337-016-3068-2

Keywords

Search

Navigation