Elsevier

Journal of Chromatography A

Volume 1313, 25 October 2013, Pages 113-118
Journal of Chromatography A

Synthesis of molecularly imprinted polymer sorbents and application for the determination of aminoglycosides antibiotics in honey

https://doi.org/10.1016/j.chroma.2013.08.072Get rights and content

Highlights

Abstract

Aminoglycoside antibiotic (AA)-selective molecularly imprinted polymer (MIP) sorbent was synthesized by polymerization of methacrylic acid and ethylene glycol dimethacrylate in the presence of streptomycin as template molecule. The MIP sorbent was in detail characterized and its performance was evaluated by selecting four model AAs including streptomycin, gentamicin, spectinomycin and dihydrostreptomycin. Relative to non-imprinted polymer (NIP), the MIPs exhibited much higher recognizable capacity and specificity to the AAs and negligible adsorption for non-AAs as well. In combination of hydrophilic interaction liquid chromatography–tandem mass spectrometry (HILIC–MS/MS), the MIP sorbent-based solid phase extraction (SPE) was able to effectively determine the residue of four model AAs in honey samples with recovery ranging from 90% to 110%. The limit of detection (and the limit of quantitation) of streptomycin, gentamicin, spectinomycin and dihydrostreptomycin was 4.5 (15.0), 2.4 (8.0), 6.0 (20.0) and 1.8 (6.0) μg/kg, respectively. Relative standard deviations of intraday and inter-day assay under three spiked levels were 4.4–12.0% and 6.8–14.6%, respectively.

Introduction

Aminoglycoside antibiotics (AAs) are a large class of antibiotics composed of amino-modified sugars, such as streptomycin, dihydrostreptomycin, spectinomycin, and gentamicin. Since their some desirable features including cheap, broad-spectrum antimicrobial and antibacterial activity, AAs have widely been used in veterinary medicine and animal husbandry to promote the growth of animals. While due to their high toxicity, high nephrotoxicity and potential hazard to human's health [1], many countries have established the maximum residue limits (MRLs) of AAs in food [2]. Thus it is necessary to develop an effective analytical method to determine AAs residue in food. Owing to their highly hydrophilic property, AAs are always poorly retained on the common C18 column or sorbent and thus it is not easy to achieve their effective separation via common C18 column. To address the issue, the separation modes based on ultra-performance liquid chromatography (UPLC) [3], ion pair chromatography (IPC) [2] and hydrophilic interaction chromatography (HILIC) [4] have been used for separation of AAs recently.

For determining trace AAs residues in food, clean-up and preconcentration procedures are usually required prior to analysis. Selective extraction of target AAs from food samples is a troublesome task since very complicated matrix of food, thus the development of novel selective sample preparation methods for AAs has attracted much interest in recent years. Among many methods, solid phase extraction (SPE) is the most widely used one owing to its some advantages such as easy operation, simplicity and low cost. C18 and weak exchange resin-based SPE have been reported for pretreating AA-contained food samples [2], [3]. Their common drawback lies in limited selectivity, for example, 72.5%, 27.4% and 64.9% of the recoveries for streptomycin, streptomycin and dihydrostreptomycin were obtained by the use of C18 resin-based SPE cartridge [3], while for weak exchange resin-based SPE, 51-76% of recoveries for 11 AAs was obtained by Babin and Fortier [5] and 85.5% and 72.3% of recoveries for streptomycin and dihydrostreptomycin was reported by Gremilogianni et al. [2].

Molecularly imprinted polymers (MIPs) are synthetic polymeric macromolecules with specific recognition sites to template molecule [6]. The polymers are prepared by the copolymerization of functional monomers and cross-linkers in the presence of template molecule. After polymerization, template molecules are removed from the polymer network, leaving the specific cavities which can selectively rebind template or template analog molecules. Other advantages of MIP include wide tolerance against extreme pH and easy preparation. These favorable features make MIP a good candidate for SPE sorbent [7], [8], [9] or solid phase microextraction coating [10], [11], [12] or monolith [13], [14] or disk-shape [15]. Recently, several kinds of MIPs have been developed for extraction of antibiotics, such as β-lactam [16], fluoroquinolone [17], tetracycline [18] and penicillin G [19]. These MIPs demonstrated satisfactory efficiency to model analysts. However, to the best of our knowledge, no report has been made for the preparation of AA-selective MIP.

The present paper describes the synthesis and characterization of AA-selective MIP by using streptomycin as the template molecule, methacrylic acid (MAA) as the functional monomer, and ethylene glycol dimethacrylate (EDMA) as the cross-linker. Four AAs, including streptomycin, dihydrostreptomycin, spectinomycin, and gentamicin (their chemical structure see Support Information (SI) of SI-Fig. 1), were selected as model analytes for evaluation of the MIP. The utility of the MIP was demonstrated by simultaneous determination of AAs residues in honey samples under the HILIC mode. Due to lack of chromosphere or fluorophore groups on AA molecule structure, common absorbance or fluorescence detector equipped by HILIC system could be not used. Here tandem mass spectrometry combined with HILIC (HILIC–MS/MS) was chosen for monitoring AAs.

Section snippets

Chemicals

Streptomycin, dihydrostreptomycin, spectinomycin and gentamicin were purchased from National Institute for the Control of Pharmaceutical and Biological Products (Shanghai, China). Alkaline aluminum oxide, anhydrous sodium sulfate, sodium chloride, sodium hydroxide and ammonium formate were obtained from Sinopharm Chemical Corp. (Shanghai, China). Formic acid, acetone, ethanol, acetonitrile, methylene chloride and hydrochloric acid were from Tianjin Chemical Reagent Company (Tianjin, China).

Removal of template molecules

The effective removal of the template molecule is a critical step in the preparation of MIP [20]. The affinity of the imprinted cavities for the template molecule makes its removal hard. This is especially true for streptomycin used here. There are strong hydrogen bonding interaction between many amino and hydroxyl groups existing on streptomycin molecule and the carboxyl groups of MAA, then leading to difficult removal of streptomycin molecule from MIP network. Much effort has been made to

Conclusions

An AA-selective MIP sorbent was synthesized using streptomycin as the template molecule. Relative to NIPs, the MIPs exhibited much higher capacity and higher selectivity to four model AAs. The MIP-based SPE in combination of HILIC–MS–MS was able to effectively determine the residue of four model AAs in honey samples. The advantage of the MIP over the common sorbents is its high selectivity to the AAs. The possible drawback of MIP lies in its lengthy preparation process, but recyclable use of

Acknowledgements

The study was financially supported by National Natural Science Foundation of China (no. 21177040, 21075038) and National Major Science Instrument Project of the Ministry of Science and Technology of China (2012YQ090229). It was partly supported by Open Foundation of Key Laboratory of Industrial Ecology and Environmental Engineering.

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