著者

Chiharu ISHII Tetsuya MIYAMOTO Shoto ISHIGO Yurika MIYOSHI Masashi MITA Hiroshi HOMMA Tadashi UEDA Kenji HAMASE

出版者

クロマトグラフィー科学会

雑誌

CHROMATOGRAPHY (ISSN:13428284)

巻号頁・発行日

pp.2017.009, (Released:2017-06-04)

参考文献数

46

被引用文献数

15

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The formation of D-amino acid residues in proteins is considered as one of the deterioration processes, and the determination of these D-amino acid residues is highly expected for the screening of new biomarkers under various disease conditions. In the present study, a two-dimensional (2D) HPLC-MS/MS system following the hydrolysis with deuterium chloride (2HCl/2H2O) and derivatization of amino acids with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) has been designed/developed and applied to the analysis of proteins stored under various conditions. As the target, 5 major D-amino acid residues (Ala, Asp, Glu, Pro and Ser) were selected. The analytical procedure was validated using a model peptide, NH2-Gly-Pro-Glu-Ala-Asp-Ser-Gly-OH, and the obtained calibration lines of %D for the 5 target amino acids were linear with correlation coefficients greater than 0.998. The RSD values for the intra-day precision and inter-day precision were lower than 5%. In most of the proteins tested, the amounts of the D-Ser and D-Asp residues increased during storage, and the highest value (14%, D-Ser) was observed in ovalbumin (OVA) after storage at pH 9.5 for 4 weeks.

著者

LIM Lee Wah

出版者

クロマトグラフィー科学会

雑誌

CHROMATOGRAPHY (ISSN:13428284)

巻号頁・発行日

pp.2015.011, (Released:2015-04-13)

参考文献数

77

被引用文献数

1 1

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This focusing review summarizes our recent works on the dissemination of capillary liquid chromatography through various approaches, i.e. simple stepwise-gradient elution system, on-line sample enrichment system, alternate-pumping recycle chromatography system, on-line immobilized-enzyme reactor, development of novel stationary phases, etc. Capillary liquid chromatography saves energy and waste, and it possesses several advantages over the conventional liquid chromatography; however, it has not been much popularized since its appearance and most users still prefer to use conventional liquid chromatography if there is a choice between them. Our research focuses on, but not limited to, developing versatile capillary liquid chromatography systems that are not only environmental-friendly but also user-friendly, and novel stationary phases that could be operated under mixed modes conditions by manipulating the eluent, which are applicable to real samples analyses will therefore eventually switch the users from “conventional” to “capillary”.

著者

Kenji HAMASE Yusuke NAKAUCHI Yurika MIYOSHI Reiko KOGA Nao KUSANO Hirohisa ONIGAHARA Hiroshi NARAOKA Hajime MITA Yasuhiko KADOTA Yasuhiro NISHIO Masashi MITA Wolfgang LINDNER

出版者

クロマトグラフィー科学会

雑誌

CHROMATOGRAPHY (ISSN:13428284)

巻号頁・発行日

vol.35, no.2, pp.103-110, 2014-08-10 (Released:2014-08-26)

参考文献数

39

被引用文献数

4 33

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A two-dimensional chiral high-performance liquid chromatographic (2D-HPLC) system has been established for the analysis of extraterrestrial amino acids. As the targets, 8 chiral amino acids (alanine (Ala), valine (Val), 2-aminobutyric acid (2AB), norvaline (nVal), N-methylalanine (N-MeAla), isovaline (iVal), 3AB and 3-aminoisobutyric acid (3AIB)) and 5 non-chiral amino acids (glycine (Gly), β-Ala, γ-aminobutyric acid (GABA), sarcosine (Sar) and 2AIB) were selected. These amino acids were tagged with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), and non-enantioselectively separated by a capillary monolithic ODS column in the first dimension. The target fractions were automatically introduced into the second dimension and further separated by Pirkle-type enantioselective columns. By using this system, the 2D-HPLC separation of 21 components in small particles of a carbonaceous chondrite (Yamato 791191, Antarctic CM2 meteorite) could be successfully performed, and all of the target amino acids were observed. The D/L ratios of the chiral molecules are almost 50/50 for all of the tested proteinogenic and non-proteinogenic amino acids.

著者

Yoshinori INOUE Atsushi YAMAMOTO

出版者

クロマトグラフィー科学会

雑誌

CHROMATOGRAPHY (ISSN:13428284)

巻号頁・発行日

vol.35, no.2, pp.63-72, 2014-08-10 (Released:2014-08-26)

参考文献数

94

被引用文献数

6

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High-performance liquid chromatography (HPLC) has great contribution in the analysis of the carbohydrates. Various separation modes such as ion exchange chromatography, size exclusion chromatography and partition chromatography have been applied to HPLC of mono- and oligosaccharides. Among them, partition chromatography with a polar stationary phase and aqueous mobile phase, called aqueous normal phase partition (ANP) chromatography, has been used in various fields, and is widely used for the separation of reducing sugars. Currently, this separation mode is considered to be a type of "hydrophilic interaction chromatography (HILIC)". As the use of HILIC mode increases, many novel stationary phases being available for the separation of reducing sugars have been developed. This article focuses on the separation of reducing sugars by ANP (HILIC) mode, and summarizes the traditional and/or recent stationary phases for the separation of reducing sugars. The applicability of various kinds of stationary phases to solid-phase extraction (SPE) of saccharides is also described.