A confirmatory method for the determination of tetracyclines in muscle using high-performance liquid chromatography with diode-array detection.
Using high-performance liquid chromatography with diode-array detection (HPLC-DAD) technique, a confirmatory method for the determination of trace levels of tetracyclines (oxytetracycline, tetracycline, chlortetracycline and doxycycline) and their 4-epimers (4-epioxytetracycline, 4-epitetracycline and 4-epichlortetracycline) in animal tissues (muscle) was developed. The samples are extracted with a mixture of succinic acid 0.1M (pH 4) and methanol after the addition of metacycline as internal standard. The clean-up is carried out by metal chelate affinity chromatography with a following concentration step on an OASIS HLB polymeric reversed phase column. The chromatographic separation of the seven analytes is achieved in 10 min on a short monolithic column (50 mm x 4.6 mm i.d.) using a gradient elution. The method was validated in bovine muscle following the Commission Decision 2002/657/EC criteria: samples spiked at four concentration levels (0.25, 0.5, 1 and 1.5 times the maximum residue limit) were analysed. Method trueness and precision (repeatability and intra-laboratory reproducibility) as well as decision limits (CCalpha) and detection capabilities (CCbeta) are reported.
Cristofani
Elisa
E
Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche, Via G. Salvemini, 1, I-06126 Perugia, Italy.
Antonini
Chiara
C
Tovo
Gloria
G
Fioroni
Laura
L
Piersanti
Arianna
A
Galarini
Roberta
R
D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis* / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004350:Drug Residues / Q000032:analysis* / Q000302:isolation & purification; D009132:Muscles / Q000737:chemistry*; D015203:Reproducibility of Results; D013754:Tetracyclines / Q000032:analysis* / Q000302:isolation & purification
D000900:Anti-Bacterial Agents; D013754:Tetracyclines