Abstract
Microchip electrophoresis (ME) was applied for the separation of two physiologically important imidazole dipeptides—carnosine and anserine. The capacitively coupled contactless conductivity detector (C4D) was employed for quantification of both dipeptides after separation in a new home-built ME unit. The separation parameters were optimized as follows to enable quantitative, baseline separation of both dipeptides: injection time 16 s, injection voltage 900 V/cm, and separation voltage 377.1 V/cm. The C4D detector responded linearly to both imidazole dipeptides in the range 0–20 mg L−1. The known addition methodology was applied to test the accuracy of the measurement of imidazole dipeptides in a complex sample. The recoveries for measurement of carnosine in the mixture ranged from 96.1 to 105.0%, whereas those for anserine amounted to 96.6 to 102.0%. This method was also applied to real biological samples. The results exhibited a satisfactory agreement with a standard HPLC method. The proposed ME method represents a cheap, fast, and simple alternative to the existing, more complicated and expensive HPLC methods. This method does not demand either the optical detectors nor tedious derivatization of sample, which are unavoidable in HPLC methods. The method was succesfuly applied for animal species determination in unknown meat samples using the carnosine/anserine ratio, and subsequently, it could be used in a food fraud prevention process.
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Acknowledgements
The authors wish to acknowledge the Centre of Excellence, Josip Juraj Strossmayer University of Osijek for providing the support of these investigations. For schematic drawings and electronic circuit schema the authors wish to thank D. Rukavina from Faculty of Electrical Engineering, J. J. Strossmayer University of Osijek.
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The animals involved in this study were managed in agreement with the European Directive 86/609/ EEC regarding the protection of animals used for experimental or other scientific purposes.
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Prof. Dr. Milan Sak-Bosnar regrettably deceased on October 17, 2017.
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Jozanović, M., Sakač, N., Sak-Bosnar, M. et al. A simple and reliable new microchip electrophoresis method for fast measurements of imidazole dipeptides in meat from different animal species. Anal Bioanal Chem 410, 4359–4369 (2018). https://doi.org/10.1007/s00216-018-1087-6
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DOI: https://doi.org/10.1007/s00216-018-1087-6