Screening method for the determination of selected tetracyclines in water by liquid chromatography with diode array detector

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A chromatographic procedure for determination of oxytetracycline (OXT), tetracycline (TC), chlorotetracycline (CTC), and doxycycline (DC) in water samples was developed and was applied for the analysis of water samples collected from poultry and pig farms and environmental water samples. Samples were acidified with trifluoroacetetic acid to pH 3 and further purified by solid phase extraction using Oasis HLB cartridges. The samples were dried up and redissolved in the mixture of oxalic acid and methanol. Separation was performed on reserved phase column (Phenomenex column C18 , 250 mm × 4.6 mm, 5 μm) by multistep gradient elution, and detection was carried out at 360 nm for OTC and TC, 370 nm for CTC, and 350 nm for DC. The tetracyclines were eluted with the mobile phase of 0.05 M oxalic acid (pH 2.5), acetonitrile, and methanol. This method provided average recoveries of 83.53% to 108.59%, with coefficient of variations (CVs) of 2.41% to 8.64% in the range of 10 to 1000 μg/L OTC, TC, CTC, and DC in water. The linearity for the tetracyclines was determined by HPLC-DAD in the range 10 to 1000 μg/L, with the correlation coefficient (R) > 0.99. The LOD and LOQ for the tetracyclines in water samples ranged from 1.51 to 4.00 and 2.51 to 5.93 μg/L, respectively.

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  • 1. Babić S. Asperger D. Mutavdzić D. Horvat A.J.M. Kastelan- Macan M.: Solid phase extraction and HPLC determination of veterinary pharmaceuticals in wastewater. Talanta 2006 70 732-738.

  • 2. Blanchflower W.J. McCracken R.J. Haggan A.S. Kennedy D.G.: Confirmatory assay for the determination of tetracycline oxytetracycline chlortetracycline and its isomers in muscle and kidney using liquid chromatography-mass spectrometry J Chromatogr B 1997 692 351-360.

  • 3. Charlet M. Schelkens M. Croubels S. De Backer P.: Quantitative multi-residue analysis of tetracyclines and their 4- epimers in pig tissues by high-performance liquid chromatography combined with positive-ion electrospray ionization mass spectrometry. Anal Chim Acta 2003 492 199-213.

  • 4. Cheng Y.F. Phillips D.J. Neue U.: Simple and rugged SPE method for the determination of tetracycline antibiotics in serum by HPLC using a volatile mobile phase. Chromatographia 1997 44 187-190.

  • 5. Cinquina A.L. Longo F. Anastasi G. Giannetti L. Cozzani R.: Validation of a high-performance liquid chromatography method for the determination of oxytetracycline tetracycline chlorotetracycline and doxycycline in bovine milk and muscle. J Chromatogr A 2003 987 227-233.

  • 6. Cooper A.D. Stubbings G.W.F. Kelly M. Tarbin J.A. Farrington W.H.H. Shearer G.: Improved method for the on-line metal chelate affinity chromatography-high-performance liquid chromatographic determination of tetracycline antibiotics in animal products. J Chromatogr A 1998 812 321-326.

  • 7. Croubels S.M. Vanoosthuyze K.E.I. Van Peteghem C.H.: Use of metal chelate affinity chromatography and membranebased ion-exchange as clean-up procedure for trace residue analysis of tetracyclines in animal tissues and egg. J Chromatogr A 1997 690 173-179.

  • 8. Furusawa N.: Isolation of tetracyclines in milk using a solidphase extracting column and water eluent. Talanta 2003 59 155-159.

  • 9. Gajda A. Posyniak A. Żmudzki J. Tomczyk G.: Determination of doxycycline in chicken fat by liqud chromatogarphy with UV detection and liquid chromatography-tandem mass spectrometry. J Chromatogr B 2013 928 113-120.

  • 10. Hirsch R. Terens T.A. Haberer K. Mehlich A. Ballwanz F. Kratz K.L.: Determination of antibiotics in different water compartments via liquid chromatography-electrospray tandem mass spectrometry. J Chromatogr A 1998 815 213-223.

  • 11. Jia A. Xiao Y. Hu J. Asami M. Kunikane S.: Simultaneoous determination of tetracyclines and their degradation products in envirnmental waters by liquid chromatography-electrospray tandem mass spectrometry. J Chromatogr A 2009 1216 4655-4662.

  • 12. Monser L. Darghouth F.: Rapid liquid chromatographic method for simultaneous determination of tetracyclines antibiotics an 6-Epi-doxycycline in pharmaceutical products using porous graphitic carbon column. J Pharm Biomed Anal 2000 23 353-362.

  • 13. Nozal L. Arce L. Simont B.M. Rios A. Valcarcel M.: Rapid determination of trace levels of tetracyclines in surface water using a continuous flow manifold coupled to a capillary electrophoresis system. In: Chemical Analysis for Antibiotics Used in Agriculture. Edited by Oka H. Nakazawa H. Harada K.I. MacNeil J.D. AOAC Inter Arlington USA 1995 p. 333.

  • 14. Pena A. Carmona A. Barbosa A. Lino C. Silveira I. Castillo B.: Determiantion of tetracycline and its major degradation products by liquid chromatography with fluorescence detection. J Chromatogr A 2003 987 227-233.

  • 15. Posyniak A. Mitrowska K. Żmudzki J. Niedzielska J.: Analytical procedure for the determination of chloroteracycline and 4-epi-chlorotetracykline in pig kidneys. J Chromatogr A 2005 1088 169-174

  • 16. Reverte S. Borrull F. Pocurull E. Marce R.M.: Determination of antibiotic compounds in water by solid-phase extraction high performance liquid chromatography (electrospray) mass spectrometry. J Chromatogr A 2003 1010 225-232.

  • 17. Rick W. Fedeniuk S.R. McCurdy A.R.: Application of reversed-phase liquid chromatography and prepacked C18 cartridges for the analysis of oxytetracycline and related compounds. J Chromatogr B 1996 677 291-297.

  • 18. Rodrigez Diaz R.C. Aguilar-Cabalos M.P. Gomez Hens A.: Simultaneous determination of ciprofloxacin and tetracyclines in biological fluids based on dual-lanthanide sensitized luminescence using dry reagent chemical technology. J Pharm Biomed Anal 2005 37 1101-1104.

  • 19. Rudek M.A. March C.L. Bauer K.S. Pluda J.M. Figg W.D.: High-performance liquid chromatography with mass spectrometry detection for quantitating COL-3 a chemically modified tetracycline in human plasma. J Pharm Biomed Anal 2000 22 1003-1014.

  • 20. Tjornelund J. Honor-Hansen S.: Use of metal complexation in non aqueous capillary electrophoresis system for the separation and improved detection of tetracyclines. J Chroamtogr A 1997 779 235-243.

  • 21. Walsh J.R. Walker L.V. Webber J.J.: Determination of tetracyclines in bovine and porcine muscle by high-performance liquid chromatography using solid-phase extraction J Chromatogr A 1992 596 211-216.

  • 22. Valverde S.R. Gil Garcia M.D. Martinez Galera M. Goicoechea H.C.: Determination of tetracyclines in surface water by partial least squares using multivariate calibration transfer to correct the effect of solid phase preconcentration in photochemically induced fluorescence signals. Anal Chim Acta 2006 562 85-93.

  • 23. Vienneau D.S. Kindberg C.G.: Development and validation of a sensitive method for tetracycline in gingival crevicular fluid by HPLC using fluorescence detection J Pharm Biomed Anal 1997 16 111-117.

  • 24. Vinas P. Balsalobre N. Lopez-Erroz C. Hernandez-Cordoba M.: Liquid chromatography with ultraviolet absorbance detection for the analysis of tetracycline residue in honey. J Chromatogr A 2004 1022 125-129.

  • 25. Yang Xio A.J. Hu J. Asami M. Kunikane S.: Simultaneous determination of tetracyclines and their degradation products in environmental waters by liquid chromatography-electrospray tandem mass spectrometry. J Chromatogr A 2009 1216 4655-4662.

  • 26. Zhu J. Snow D.D. Cassada D.A. Monson S.J. Spalding R.F.: Analysis of oxytetracycline tetracycline and chlorotetracycline in water using solid-phase extraction and liquid chromatography tandem mass spectrometry. J Chromatogr A 2001 928 177-186.

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