Determination of fluoroquinolones in chicken feces - a new liquid-liquid extraction method combined with LC-MS/MS.

Determination of fluoroquinolones in chicken feces - a new liquid-liquid extraction method combined with LC-MS/MS.

Environmental toxicology and pharmacology (2014-10-13)

Franziska Janusch, Gesine Scherz, Siegrun A I Mohring, Gerd Hamscher

ABSTRACT

The application of antibiotics including fluoroquinolones to farming animals is widespread and may lead to the development of antibiotic resistance and other environmental effects. To calculate environmental loads and for a proper risk assessment it is necessary to determine the antibiotic concentration in feces. Therefore, a new liquid-liquid extraction method combined with HPLC-MS/MS for the detection of marbofloxacin, ciprofloxacin, enrofloxacin and difloxacin in chicken feces was developed. Recoveries ranged from 51.0% to 83.5%. LOQs were between 0.10 and 1.09μg/kg. Feces of chickens treated with an enrofloxacin dosage of 10mg/kg bodyweight revealed maximum enrofloxacin and ciprofloxacin concentrations of 61.3 and 18.8mg/kg. Both antibiotics could be detected in feces up to two days after the last application in notable amounts (∼1mg/kg). Thus, feces of recently medicated chickens should not be used as a fertilizer without any further processing.

MATERIALS

Product Number

Brand

Product Description

79617

Sigma-Aldrich

Phosphoric acid, BioUltra, ≥85% (T)

03609

Sigma-Aldrich

Ethylenediaminetetraacetic acid, BioUltra, ≥99.0% (KT)

W248703

Sigma-Aldrich

Formic acid, ≥95%, FCC, FG

W241407

Sigma-Aldrich

Ethyl acetate, ≥99%, FCC, FG

W241415

Sigma-Aldrich

Ethyl acetate, natural, ≥99%, FCC, FG

02575

Supelco

Dichloromethane, analytical standard

17850

Sigma-Aldrich

Ciprofloxacin, ≥98% (HPLC)

58958

Supelco

Ethyl acetate, analytical standard

60353

Sigma-Aldrich

Potassium phosphate dibasic, anhydrous, suitable for luminescence, Molecular Biology, BioUltra, ≥99.0% (T)

06473

Sigma-Aldrich

Formic acid solution, BioUltra, 1.0 M in H₂O

17849

Sigma-Aldrich

Enrofloxacin, ≥99.0%

91237

Supelco

2-Propanol, analytical standard

295426

Sigma-Aldrich

Hydrogen chloride, ReagentPlus^®, ≥99%

270989

Sigma-Aldrich

Ethyl acetate, anhydrous, 99.8%

270997

Sigma-Aldrich

Dichloromethane, anhydrous, ≥99.8%, contains 40-150 ppm amylene as stabilizer

59304

Sigma-Aldrich

2-Propanol, BioUltra, Molecular Biology, ≥99.5% (GC)

03690

Sigma-Aldrich

Ethylenediaminetetraacetic acid disodium salt solution, BioUltra, pH 8.0, ~0.5 M in H₂O

537446

Sigma-Aldrich

Ethyl acetate, ReagentPlus^®, ≥99.8%

675431

Sigma-Aldrich

2-Propanol, HPLC Plus, for HPLC, GC, and residue analysis, 99.9%, poly coated bottles

466123

Sigma-Aldrich

Phosphoric acid, crystalline, ≥99.999% trace metals basis

345245

Sigma-Aldrich

Phosphoric acid, 85 wt. % in H₂O, 99.99% trace metals basis

278475

Sigma-Aldrich

2-Propanol, anhydrous, 99.5%

439223

Sigma-Aldrich

Dichloromethane, suitable for HPLC, ≥99.9%, contains 40-150 ppm amylene as stabilizer

431788

Sigma-Aldrich

Ethylenediaminetetraacetic acid, 99.995% trace metals basis

450200

Sigma-Aldrich

Potassium phosphate dibasic, 99.95% trace metals basis

452289

Sigma-Aldrich

Phosphoric acid, ≥85 wt. % in H₂O, ≥99.999% trace metals basis

S3139

Sigma-Aldrich

Sodium phosphate monobasic, BioReagent, Molecular Biology, anhydrous, ≥98%

EDS

Sigma-Aldrich

Ethylenediaminetetraacetic acid, BioUltra, anhydrous, ≥99% (titration)

E9884

Sigma-Aldrich

Ethylenediaminetetraacetic acid, ACS reagent, 99.4-100.6%, powder

S5011

Sigma-Aldrich

Sodium phosphate monobasic, BioPerformance Certified, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0% (titration)