Skip to main content
SpringerLink
Log in

Determination of Levamisole in Sheep Muscle Tissue by High-Performance Liquid Chromatography and Photo Diode Array Detection

  • Published:
Chromatographia Aims and scope Submit manuscript

Abstract

A high-performance liquid chromatographic method for the determination of levamisole (LVM) residues in sheep muscle tissue is described. LVM was extracted with ethyl acetate under alkaline conditions and cleanup was performed by liquid-liquid partition between organic-basic and organic-acid medium. Finally, levamisole was back extracted with chloroform carefully transferred into a clean glass vial and evaporated to dryness at 50 °C under a gentle stream of nitrogen. The remaining dry residue was dissolved in the mobile phase used, filtered and an aliquot was injected automatically into the chromatograph for analysis. Chromatography was performed on a Zorbax®SB-C18 column at 50 °C and detection by a PDA detector monitored at λ max 220 nm. The mobile phase was a mixture of 0.1 % trifluoroacetic acid (v/v) pH 2.0 and acetonitrile-methanol 3 : 2 (v/v) in a combination of 30 : 70 (v/v) and a flow rate of 1.0 mL min−1, delivered isocratically. This analytical method was validated by assessing recovery efficiency using spiked muscle tissue samples with standard solutions in methanol at four fortification levels of 1/2 MRL, 1 MRL, 2 MRL and 4 MRL and five times for each concentration (n = 5). Mean recovery (R%) achieved for muscle tissue was 75.65 ± 2.74% with an acceptable Relative Standard Deviation RSD% = 10.4. The same method was used also for the analysis of kidney, liver and fat (perirenal) and the recoveries found were 70.25 ± 1.07% (RSD% = 1.52), 72.37 ± 3.6% (RSD% = 4.97) and 69.44 ± 2.22% (RSD% = 3.19), respectively. The limit of detection (LOD) for muscle tissue was found to be 2.0 µg kg−1 and the limit of quantification (LOQ) 5.0 µg kg−1.

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

Similar content being viewed by others

References

  • Braund KG (2003) In: Clinical Neurology in Small Animals-Localization, Diagnosis and Treatment, Braund KG (Ed.), Ithaca. International Veterinary Information Service, USA, pp. 1–23

  • Cherlet M, de Baere S, Croubels S, de Backer P (2000) J Chromatogr B 742:283–293

    CAS  Google Scholar 

  • Sahagun AM, Teran MT, Garcia JJ, Fernandez N, Sierra M, Diez MJ (2001) J Vet Pharmacol Ther 24:439–442

    Article  CAS  Google Scholar 

  • IPCS (1991) In: Toxicological evaluation of certain veterinary drug residues in food. The thirty-sixth meeting of the joint FAO/WHO expert committee on food additives (JECFA). WHO Food additives series: 27. International Programme on Chemical Safety. World Health Organization, Geneva, pp. 75–101

    Google Scholar 

  • Dolezalova M. Tkaczykova M (2001) J Pharm Biomed Anal 25:407–415

    Google Scholar 

  • Chankvetadzea B, Burjanadze N, Santi M, Massolini G, Blaschke G (2002) J Sep Sci 25:733–740

    Article  Google Scholar 

  • Woestenborghs R, Michielsen L, Heykants J (1981) J Chromatogr, 224:25–32

    Google Scholar 

  • de Bukanski BW, Degroodt JM, Beernaert HZ (1991) Z Lebensm Unters Forsch A 193:545–547

    Article  Google Scholar 

  • Vandamme TF, Demoustier M, Rollmann B (1995) Eur J Drug Metab Pharmacokinet 20:145–149

    CAS  Google Scholar 

  • Dreassi E, Corbini G, La Rosa C, Politi N, Corti P (2001) J Agric Food Chem 49:5702–5705

    Article  CAS  Google Scholar 

  • De Ruyck H, Daeseleire E, de Rider H, Van Renterghem R (2002) J Chromatogr A 976:181–194

    Article  Google Scholar 

  • Edder P, Ortelli D, Jan P, Corvi C (2004). In EURORESIDUE V Conference on Residues of Veterinary Drugs in Food. 1st ed. van Gingel LA, Bergwerff AA (ed.) Colophon, Noordwijkerhout, The Netherlands, pp. 466–469

  • De Ruyck H, Van Renterghem R, De Ridder H, De Brabander D (2000) Food Control 11:165–173

    Article  Google Scholar 

  • COMMISSION REGULATION (EC) No 508/1999 of 4 March 1999 amending Annexes I to IV to Council Regulation (EEC) No 2377/90 laying down a Community procedure for the establishment of maximum residue limits of veterinary medicinal products in foodstuffs of animal origin. OJEC L 60/16 9. 3. 1999.

  • Tyrpenou AE, Tsigouri AD, Gouta-Papastavrou E (1995) In: Proceedings of the Greek Congress “Food quality and consumers' demands beyond 2000”. DETROP '95 Thessaloniki, Greece, pp. 34–35

  • Tyrpenou AE, Rigos G (2004) Chromatographia 60:657–661

    Article  CAS  Google Scholar 

  • El-Kholy H, Kemppainen BW (2003) J Chromatogr B 796:371–377

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratory of High Performance Liquid Chromatography, Veterinary Research Institute of Thessaloniki, National Agricultural Research Foundation, NAGREF Campus, 570 01, Thermi, Thessaloniki, Greece

    A. E. Tyrpenou

  2. Department of Anatomy & Physiology of Farm Animals, Faculty of Animal Science, Agricultural University of Athens, 75, Iera Odos Street, 118 55, Athens, Greece

    E. M. Xylouri-Frangiadaki

Authors
  1. A. E. Tyrpenou
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E. M. Xylouri-Frangiadaki
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to A. E. Tyrpenou.

Additional information

Revised: 4 and 24 January 2006

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tyrpenou, A.E., Xylouri-Frangiadaki, E.M. Determination of Levamisole in Sheep Muscle Tissue by High-Performance Liquid Chromatography and Photo Diode Array Detection. Chroma 63, 321–326 (2006). https://doi.org/10.1365/s10337-006-0754-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1365/s10337-006-0754-5

Keywords

Search

Navigation