TY - JOUR T1 - Capillary electrophoresis of phosphorylated amino acids with fluorescence detection. AU - Zhang,Li-Yao, AU - Sun,Meng-Xiang, Y1 - 2007/09/22/ PY - 2007/07/10/received PY - 2007/08/25/revised PY - 2007/09/02/accepted PY - 2007/10/10/pubmed PY - 2008/1/31/medline PY - 2007/10/10/entrez SP - 30 EP - 6 JF - Journal of chromatography. B, Analytical technologies in the biomedical and life sciences JO - J Chromatogr B Analyt Technol Biomed Life Sci VL - 859 IS - 1 N2 - A rapid and sensitive capillary electrophoresis (CE) method coupled with fluorescence detection was developed for identification of protein phosphorylation by determination of phosphoamino acids. Naphthalene-2,3-dicarboxaldehyde (NDA), a fluorescence derivatization reagent, was used to label protein hydrolysate. The optimal derivatization reaction was performed with 3.5mM NDA, 40 mM NaCN and 20mM borate buffer (pH 10.0) for 15 min. The baseline separation of three phosphorylated amino acids could be obtained in less than 180 s with good repeatability by using 30 mM borate (pH 9.2) containing 2.0mM beta-cyclodextrin (beta-CD) as the running buffer. The detection limits for phosphothreonine, phosphotyrosine and phosphoserine were 7.0 x 10(-9)M, 5.6 x 10(-9)M and 7.2 x 10(-9)M, respectively (S/N=3). Also, the interference from other protein amino acids with large molar excess over that of phosphoamino acids was studied. With beta-casein as the analysis protein, this method was successfully validated. SN - 1570-0232 UR - https://www.unboundmedicine.com/medline/citation/17923447/Capillary_electrophoresis_of_phosphorylated_amino_acids_with_fluorescence_detection_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1570-0232(07)00650-2 DB - PRIME DP - Unbound Medicine ER -