Rapid analysis of amino acids in Japanese green tea by microchip electrophoresis using plastic microchip and fluorescence detection.
Microchip electrophoresis for the short-time analysis of amino acids in Japanese green tea was developed. The amino acids in Japanese green tea were derivatized with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F). The derivatives were filtered and directly analyzed by electrophoresis on a plastic microchip with a 31-mm long separation channel with fluorescence detection. Amino acid analysis of Japanese green tea was improved by removing polyphenols using a polyvinylpolypyrrolidone pretreatment. Elution profiles of NBD-amino acids were examined under different running buffer conditions, and the sodium dodecyl sulphate in the running buffer exhibited a dramatically high-separation efficiency of amino acids by inhibiting their adsorption on the channel walls. Under the optimized conditions (5 mM phosphate buffer (pH 5.5) containing 0.05 mM sodium dodecylsulfate as running buffer), the main amino acids contained in Japanese green tea were well separated within 2 min, and theanine (1475 mg/100 g tea leaf), Arg (408 mg/100 g tea leaf) and Gln (217 mg/100 g tea leaf) were detected in Japanese green tea.
Kato
Masaru
M
Department of Analytical Chemistry, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada Shizuoka, Shizuoka 422-8526, Japan.
Gyoten
Yukari
Y
Sakai-Kato
Kumiko
K
Toyo'oka
Toshimasa
T
D000097:Acetonitriles / Q000737:chemistry; D000596:Amino Acids / Q000032:analysis*; D002021:Buffers; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D008904:Miniaturization; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D013662:Tea / Q000737:chemistry*
D000097:Acetonitriles; D000596:Amino Acids; D002021:Buffers; D013662:Tea; D012967:Sodium Dodecyl Sulfate; C032159:acetonitrile