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

PHR1167

Supelco

Ciprofloxacin, Pharmaceutical Secondary Standard; Certified Reference Material

466123

Sigma-Aldrich

Phosphoric acid, crystalline, ≥99.999% trace metals basis

537446

Sigma-Aldrich

Ethyl acetate, ReagentPlus^®, ≥99.8%

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

437549

Sigma-Aldrich

Ethyl acetate, ACS reagent, ≥99.5%

270989

Sigma-Aldrich

Ethyl acetate, anhydrous, 99.8%

295426

Sigma-Aldrich

Hydrogen chloride, ReagentPlus^®, ≥99%

278475

Sigma-Aldrich

2-Propanol, anhydrous, 99.5%

439223

Sigma-Aldrich

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

79617

Sigma-Aldrich

Phosphoric acid, BioUltra, ≥85% (T)

609781

Sigma-Aldrich

Phosphoric acid-¹⁶O₄ solution, 70 wt. % in D₂O, 99.9 atom % ¹⁶O

609765

Sigma-Aldrich

Sodium hydroxide-¹⁶O solution, 20 wt. % in H₂¹⁶O, 99.9 atom % ¹⁶O

E8008

Sigma-Aldrich

Ethylenediaminetetraacetic acid solution, 0.02% in DPBS (0.5 mM), sterile-filtered, BioReagent, suitable for cell culture

345245

Sigma-Aldrich

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

270997

Sigma-Aldrich

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

437522

Sigma-Aldrich

2-Propanol, ACS reagent, ≥99.5%

PHR1072

Supelco

2-Propanol, Pharmaceutical Secondary Standard; Certified Reference Material

24233

Sigma-Aldrich

Dichloromethane, puriss., meets analytical specification of Ph. Eur., NF, ≥99% (GC)

494518

Sigma-Aldrich

Ethyl acetate, biotech. grade, ≥99.8%

673773

Sigma-Aldrich

2-Propanol, ACS reagent, ≥99.5%

695076

Sigma-Aldrich

Formic acid, ACS reagent, ≥96%

27001

Sigma-Aldrich

Formic acid, puriss., meets analytical specifications of DAC, FCC, 98.0-100%

30417

Sigma-Aldrich

Phosphoric acid, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥85%

27227

Sigma-Aldrich

Ethyl acetate, puriss., meets analytical specification of Ph. Eur., BP, NF, ≥99.5% (GC)

320269

Sigma-Aldrich

Dichloromethane, ACS reagent, ≥99.5%, contains 40-150 ppm amylene as stabilizer

33015

Sigma-Aldrich

Formic acid, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥98%

33539

Sigma-Aldrich

2-Propanol, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.8% (GC)

24137

Sigma-Aldrich

2-Propanol, puriss., meets analytical specification of Ph. Eur., BP, USP, ≥99.5% (GC)

L092003

Sigma-Aldrich

Ethyl acetate