Derivative spectrum chromatographic method for the determination of trimethoprim in honey samples using an on-line solid-phase extraction technique |
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Authors: | Uchiyama Kazuhisa Kondo Mari Yokochi Rika Takeuchi Yuri Yamamoto Atsushi Inoue Yoshinori |
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Affiliation: | College of Bioscience and Biotechnology, Chubu University, Kasugai-shi, Aichi, Japan. |
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Abstract: | A simple, selective and rapid analytical method for determination of trimethoprim (TMP) in honey samples was developed and validated. This method is based on a SPE technique followed by HPLC with photodiode array detection. After dilution and filtration, aliquots of 500 μL honey samples were directly injected to an on-line SPE HPLC system. TMP was extracted on an RP SPE column, and separated on a hydrophilic interaction chromatography column during HPLC analysis. At the first detection step, the noise level of the photodiode array data was reduced with two-dimensional equalizer filtering, and then the smoothed data were subjected to derivative spectrum chromatography. On the second-derivative chromatogram at 254 nm, the limit of detection and the limit of quantification of TMP in a honey sample were 5 and 10 ng/g, respectively. The proposed method showed high accuracy (60-103%) with adequate sensitivity for TMP monitoring in honey samples. |
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Keywords: | Derivative spectrum chromatography Honey HPLC‐photodiode array On‐line SPE Trimethoprim |
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