Development of a rapid method for the determination of antibiotic residues in honey using UPLC-ESI-MS/MS

Abstract

An accurate, reliable and fast multianalyte/multiclass ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the simultaneous analysis of 23 pharmaceuticals, belonging to different classes amphenicols, sulfonamides, tetracyclines, in honey samples. The method developed consists of ultrasonic extraction followed by UPLC-ESI-MS/MS with electrospray ionization in both positive mode and negative mode. The influence of the extraction solvents and mobile phase composition on the sensitivity of the method, and the optimum conditions for sample weight and extraction temperature in terms of analyte recovery were extensively studied. The identification of antibiotics is fulfilled by simultaneous use of chromatographic separation using an Acquity BEH C 18 (100 mm x 2.1 mm, 1.7 µm) analytical column with a gradient elution of mobile phases and tandem mass spectrometry with an electrospray ionization. Finally, the method developed was applied to the determination of target analytes in honey samples obtained from the local markets and several beekeepers in Muğla, Turkey. Ultrasonic-extraction of pharmaceuticals from honey samples is a well-established technique by UPLC-ESI-MS/MS, the uniqueness of this study lies in the simultaneous determination of a remarkable number of compounds belonging to 23 drug at the sub-nanogram per kilogram level.Keywords: antibiotics; fast method; honey; UPLC-ESI-MS/MS; chloramphenicol.Practical Application: Honey is one of the healthy foods across the world. Food safety is an important issue regarding residues of pharmaceutical drugs. The developed method proposed simultaneous analysis of analysis of twenty-three veterinary pharmaceuticals in honey. A fast and sensitive multianalyte/multiclass ultra-performance liquid chromatography-tandem mass spectrometry with electrospray ionization (UPLC-ESI-MS/MS) method provides the analysis of 50 honey samples in 50 minutes for food and laboratory sectors, and benefits safe food consumption.

Residue Determination in Honey

“…Moreover, after derivatization with fluorescamine, sulphonamides give fluorescence and some procedures apply LC-FLD (with pre-or post-column derivatization), reaching limits of detections (LOD/CCβ) comparable to those of LC-MS methods. In Table 9, some example of these applications are listed [69][70][71][72][73][74][75][76][77][78]. Since considerable amounts of SAs are bound to honey sugars, in 2000 Schwaiger and Schuch [79] demonstrated the need of an acidic hydrolysis prior to the residue analysis.…”

“…Therefore, at one hand, exploiting their basic moiety, some procedures use strong cation exchange (SCX-SPE) to isolate sulphonamides from the acidic honey extracts [71][72][73]77]. On the other hand, to successfully apply RP-SPE or LLE clean-up, some researchers buffered honey extract in the pH range about 4-6 in which the neutral form of sulphonamides prevails [70,[74][75][76]78]. In this interval, the distribution coefficients (D) reach their maximum and the compound lipophilicity is enhanced, as shown in Figure 3 for sulphathiazole.…”

A genetically engineered Escherichia coli that senses and degrades tetracycline antibiotic residue

“…Antibiotics are widely used in hospitals, animal husbandry and aquaculture industry to prevent bacterial infections and to boost growth. As a result, antibiotics remain in meat products [ [1] , [2] , [3] , [4] , [5] ], honey [ [5] , [6] , [7] , [8] , [9] , [10] , [11] , [12] , [13] , [14] ] and milk supplies throughout the world [ [15] , [16] , [17] , [18] , [19] , [20] ]. Waste water from above facilities is emitted into the environment [ 21 ] without proper treatment, thus leading to a large amount of environmental antibiotic residues.…”

Development and Validation of a Simple and Fast Method for Sulfonamides, Tetracyclines and Macrolides in Honey Using Lc-MS/MS

“…The use of antibacterials in foodproducing animals may leave residues in foodstuffs of animal origin like meat, milk, honey and eggs (Lee et al, 2001; Kibroyesfa and Naol, 2017). Honey is one of the healthy food across the world used for thousands of years in world-wide, being rich in minerals, antioxidants, and simple sugars (Kivrak et al, 2016). Sulfonamides, tetracyclines, nitrofurans and macrolides are used to prevent and combat diseases in bees, and the use of these substances has been known since 1940 (Benetti et al, 2006; Barganska et al, 2011).…”

“…Several methods were described in the literature about antibacterial residues in honey, but there are few reports for multiclass methods for this matrix. Kivrak et al (2016), developed a method for amphenicols and sulfonamides in honey, using dissolution with acetic acid 0.5% and ultrasonic bath at 50 C for 30 minutes. The analysis of sulfonamides and chloranphenicol in honey was developed, using Solid-Phase Extraction (SPE) (Sheridan et al, 2008).…”