A New strategy for Quantitative Analysis of Ergothioneine in Fermentation Broth by RP-HPLC
An efficient and sensitive method was established for quantitative analysis of L-Ergothioneine during fermentation analyze by reversed-phase-high performance liquid chromatography (RP-HPLC). The method was carried out on two C18 columns (4.6 × 250 mm, 5 μ
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A New strategy for Quantitative Analysis of Ergothioneine in Fermentation Broth by RP-HPLC Tao Zhou, Qi Liu, Wenxia Jiang and Ning Chen
Abstract An efficient and sensitive method was established for quantitative analysis of L-Ergothioneine during fermentation analyze by reversed-phase-high performance liquid chromatography (RP-HPLC). The method was carried out on two C18 columns (4.6 9 250 mm, 5 lm), and the isocratic mobile phase was 1 % methanol containing boric acid adjusted to a pH of 5.0 with a flow rate of 0.7 mL/ min. An UV–VIS detector equipped with a wavelength of 257 nm was employed. The injection volume was 5 lL, with the columns temperature being 25 °C. The linearity, recovery, limit of detection (LOD) and quantification (LOQ), precision, repeatability, stability, and recovery were all tested and good results were obtained. The method was simple, rapid, accurate, and high sensitivity and could be utilized for the research and development of L-Ergothioneine in industry. Keywords RP-HPLC
L-Ergothioneine Fermentation broth
32.1 Introduction L-Ergothioneine (2-Thiol-L-histidine-betaine, EGT) is a naturally occurring chiral amino acid biosynthesized in some bacteria and fungi but not in higher plants and animals [1]. A great number of rationales have been proposed to explain the universal presence of EGT and its accumulation and function in tissues, including T. Zhou N. Chen College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300222, China T. Zhou Q. Liu W. Jiang (&) Tianjin Key Laboratory for Industrial Biological Systems and Bioprocessing Engineering, Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China e-mail: [email protected]
T.-C. Zhang et al. (eds.), Proceedings of the 2012 International Conference on Applied Biotechnology (ICAB 2012), Lecture Notes in Electrical Engineering 249, DOI: 10.1007/978-3-642-37916-1_32, Ó Springer-Verlag Berlin Heidelberg 2014
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a role for EGT as a factor in bioenergetics [2], immune regulator [3, 4], regulator of gene expression [5], divalent metal chelator [6–8], and ultraviolet ray filter [9], while its most important function is as an physiologic antioxidant and physiologic cytoprotectant [10–16]. EGT was first isolated from ergot fungus that devastated rye grain [17]. In aqueous solution, EGT exists as a thione-thiol tautomer, with the thione form predominating at physiological pH [18]. Therefore, EGT can instead other antioxidants (such as glutathione) and antiseptics due to its nontoxic and stability and would be applied to organ transplantation technology, cell preservation technology, pharmaceuticals industry, food software industry, functional food industry, animal feeding stuffs industry, cosmetic industry, biotechnology, and so on. There were several methods for assaying EGT including chemical assay, spectrophotometry [19], and HPLC [20]. Currently, methods that utilize spectrophotometric analysis t
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