Determination of Residual Fluoroquinolones in Honey by Liquid Chromatography Using Metal Chelate Affinity Chromatography

Abstract

A new analytical method for the simultaneous determination of seven fluoroquinolones, namely, norfloxacin, ciprofloxacin, danofloxacin, enrofloxacin, orbifloxacin, sarafloxacin, and difloxacin, especially in dark-colored honey, has been developed. Fluoroquinolone antibiotics were extracted from samples with MacIlvaine buffer solution (pH 4.0) containing EDTA disodium salt dihydrate. The extracts were treated with both a polymeric cartridge and a metal chelate affnity column preloaded with ferric ion (Fe3+). LC separation with fluorescence detection was performed at 40°C using an Inertsil ODS-4 analytical column (150 × 4.6 mm, 3 µm). The mobile phase was composed of 20 mM/L citrate buffer solution (pH 3.1)–acetonitrile mixture (70 + 30, v/v) containing 1 mM/L sodium dodecyl sulfate. Lomefloxacin was used as an internal standard. The developed method was validated according to the criteria of European Commission Decision 2002/657/EC. Decision limits and detection capabilities were below 2.9 and 4.4 μg/kg, respectively.

Residue Determination in Honey

“…Thanks to the high sensitivity of fluorescence detection and the lower cost of equipment compared to LC-MS, this technique is still used to detect and confirm quinolone residues in food. In Table 7, the most significant methods are listed [52][53][54][55][56][57][58]. Generally, the solubilized honey is purified by reversed-phase SPE [53,54] or by LLE [55][56][57].…”

“…In Table 7, the most significant methods are listed [52][53][54][55][56][57][58]. Generally, the solubilized honey is purified by reversed-phase SPE [53,54] or by LLE [55][56][57]. SPE sorbents, other than reversed-phase types, are reported in the papers published in 1998 by Rose et al [52] and in 2011 by Yatsukawa et al [54].…”

“…Generally, the solubilized honey is purified by reversed-phase SPE [53,54] or by LLE [55][56][57]. SPE sorbents, other than reversed-phase types, are reported in the papers published in 1998 by Rose et al [52] and in 2011 by Yatsukawa et al [54]. Rose et al describe two parallel sample treatment protocols using ion-exchange solid-phase extraction: one for nine amphoteric QNs (ciprofloxacin, danofloxacin, enoxacin, enrofloxacin, lomefloxacin, marbofloxacin, norfloxacin, ofloxacin and sarafloxacin) and another for three acidic ones (flumequine, nalidixic acid and oxolinic acid).…”

“…This is probably the only published application of MCAC to quinolone purification, exploiting their chelating properties. The achievable selectivity allows an efficient removal of interferences also in dark-coloured honey samples such as manuka and buckwheat [54]. On the other hand, MCAC is a well-known stationary phase to purify tetracycline antibiotics using copper (Cu 2+ ) as metal ion (see Section 5.9).…”

Analysis of danofloxacin, difloxacin, ciprofloxacin and sarafloxacin in honey using micellar liquid chromatography and validation according to the 2002/657/EC decision

“…26 However, only a few studies have been published about the analysis of quinolones in honey using LC-FLD. 27,28 Furthermore, most of the extraction procedures applied to analyze honey require clean-up procedures that are tedious and time-consuming, because of the viscosity and the presence of a large amount of sugars. The most usual methods are liquid/liquid 16,18,19 or solid/liquid 16,20,27,28 extraction, or precipitation of matrix compounds.…”