title: Simultaneous determination of homocysteine, methionine and cysteine in maternal plasma after delivery by HPLC-fluorescence detection with DBD-F as a label.
abstract: An HPLC-fluorescence (FL) method for determination of sulfur-containing amino acids such as homocysteine (Hcy), methionine (Met) and cysteine (Cys) in human plasma was developed. The sulfur-containing amino acids were labeled with 4-(N,N-dimethylaminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole (DBD-F). Calibration curves in the range of 1-100 µm (Hcy and Met) and 5-500 µm (Cys) indicated good linearities (r ≥ 0.998). The limits of detection at a signal-to-noise ratio of 3 were 0.13 (Hcy), 0.02 (Met) and 0.11 µm (Cys), respectively. Acceptable results for accuracy and precision of intra- and inter-day measurements were obtained. The results of Hcy and Cys obtained by the proposed method indicated good correlations with the conventional method (r > 0.911, n = 20). Furthermore, the method was applied to determination of the sulfur-containing amino acids in maternal plasma (n = 200) after delivery. The concentrations of Hcy, Met and Cys as a median (inter quartile range, Q1 and Q3 ) were 5.37 (3.32-7.79) μm, 25.20 (20.10-31.06) μm and 147.25 (102.81-189.31) μm, respectively.
authors.affiliation: Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki, 852-8521, Japan.
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Wada
authors.forename: Maki
authors.initials: M
authors.lastname: Hirose
authors.forename: Mana
authors.initials: M
authors.lastname: Kuroki
authors.forename: Rie
authors.initials: R
authors.lastname: Ikeda
authors.forename: Yui
authors.initials: Y
authors.lastname: Sekitani
authors.forename: Noboru
authors.initials: N
authors.lastname: Takamura
authors.forename: Naotaka
authors.initials: N
authors.lastname: Kuroda
authors.forename: Kenichiro
authors.initials: K
authors.lastname: Nakashima
mesh_terms: D000328:Adult; D000603:Amino Acids, Sulfur / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D016014:Linear Models; D010080:Oxazoles / Q000737:chemistry*; D011181:Postnatal Care; D049590:Postpartum Period / Q000097:blood*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D013438:Sulfhydryl Compounds / Q000097:blood; D013449:Sulfonamides / Q000737:chemistry*
chemical_list: D000603:Amino Acids, Sulfur; D005456:Fluorescent Dyes; D010080:Oxazoles; D013438:Sulfhydryl Compounds; D013449:Sulfonamides; C060548:4-(N,N-dimethylaminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole