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Introduction
Honey bees are easily attacked by microorganisms and are highly susceptible to infections caused due to parasitic mites, fungi, bacteria and numerous other viruses due to their very weak immune system. Continuous attack of these pathogens induces a lot of stress to honey bees which results in the reduction of bee colonies worldwide. So, generally veterinary drugs are administered to honey bees, to prevent it from diseases and to limit the losses in apiculture1.But on the other hand due to the resistance of diseases against certain veterinary drugs, the veterinary treatments are administered to bee colonies at huge amounts for a longer period of time which results in leaving the traces of these drug residues in honey2.
Among all the veterinary drugs, Sulfonamides contribute to major portion as they have superior curative properties and also easily available3. Traces of these veterinary drug residues in food system causes a lot of negative health effects such as antimicrobial drug resistance, disruption of intestinal microflora , carcinogenety etc4,5,6. Due to this, Food safety and standards (contaminants, toxins and residues) regulations, 2011(India) has recommended the maximum limit of quantification when determined by LC-MS/MS (Liquid Chromatography- Tandem Mass Spectrometry) shall not exceed 5µg/kg for sulfonamide and its metabolites7.
LC-MS/MS is one of the most widely used analytical methods for the determination of veterinary drug residues in food systems8. Generally, sample preparation techniques based on solvent extraction has been highly reported in honey samples9,10. Recently papers have been published on increasing the efficiency of sample preparation conditions through ultrasonication. The low frequency sound waves create mechanical energy in the form of shear and cavitation which helps in increasing the recovery rate11-13. The present work reports a sensitive method developed for the determination of Sulfonamides in honey samples. The main objective here is to optimise the sample preparation conditions such that the optimised conditions can provide better extraction efficiency and yield higher recovery.
Material and methods
Chemicals and Materials
Veterinary drug standards (Sulfamethizole, Trimethoprim) and formic acid were purchased from Sigma (Dorset, UK). Methanol was supplied by Merck (Darmstadt, Germany). Bondesil- PSA (primary-secondary amine) and C18 endcapped bulk sorbents were supplied by Agilent, Santa Clara,...