title: Quantitative mass spectrometric analysis of dipeptides in protein hydrolysate by a TNBS derivatization-aided standard addition method.
abstract: The aim of this study was to establish, through a standard addition method, a convenient quantification assay for dipeptides (GY, YG, SY, YS, and IY) in soybean hydrolysate using 2,4,6-trinitrobenzene sulfonate (TNBS) derivatization-aided LC-TOF-MS. Soybean hydrolysate samples (25.0 mg mL(-1)) spiked with target standards were subjected to TNBS derivatization. Under the optimal LC-MS conditions, five target dipeptides derivatized with TNBS were successfully detected. Examination of the standard addition curves, with a correlation coefficient of r(2) > 0.979, provided a reliable quantification of the target dipeptides, GY, YG, SY, YS, and IY, in soybean hydrolysate to be 424 ± 20, 184 ± 9, 2188 ± 199, 327 ± 16, and 2211 ± 133 μg g(-1) of hydrolysate, respectively. The proposed LC-MS assay is a reliable and convenient assay method, with no interference from matrix effects in hydrolysate, and with no requirement for the use of an isotope labeled internal standard.
authors.affiliation: Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School of Kyushu University, 6-10-1 Hakozaki, Fukuoka 812-8581, Japan.
authors.forename: Vu Thi
authors.initials: VT
authors.lastname: Hanh
authors.affiliation: Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School of Kyushu University, 6-10-1 Hakozaki, Fukuoka 812-8581, Japan.
authors.forename: Yutaro
authors.initials: Y
authors.lastname: Kobayashi
authors.affiliation: FUJI OIL CO., Ltd., Research and Development Division, 4-3, Kinunodai, Tsukuba, Ibaraki 300-2497, Japan.
authors.forename: Motohiro
authors.initials: M
authors.lastname: Maebuchi
authors.affiliation: FUJI OIL CO., Ltd., Research and Development Division, 4-3, Kinunodai, Tsukuba, Ibaraki 300-2497, Japan.
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Nakamori
authors.affiliation: Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School of Kyushu University, 6-10-1 Hakozaki, Fukuoka 812-8581, Japan.
authors.forename: Mitsuru
authors.initials: M
authors.lastname: Tanaka
authors.affiliation: Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School of Kyushu University, 6-10-1 Hakozaki, Fukuoka 812-8581, Japan. Electronic address: tmatsui@agr.kyushu-u.ac.jp.
authors.forename: Toshiro
authors.initials: T
authors.lastname: Matsui
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods; D004151:Dipeptides / Q000032:analysis*; D007554:Isotopes; D013058:Mass Spectrometry / Q000379:methods*; D011492:Protein Hydrolysates / Q000032:analysis*; D013025:Glycine max / Q000737:chemistry*; D014302:Trinitrobenzenesulfonic Acid / Q000031:analogs & derivatives
chemical_list: D004151:Dipeptides; D007554:Isotopes; D011492:Protein Hydrolysates; D014302:Trinitrobenzenesulfonic Acid
keywords: Dipeptide; LC–TOF-MS; Standard addition method; TNBS derivatization