Analysis of O-Phosphoamino Acids in Biological Samples by Gas Chromatography with Flame Photometric Detection

Kataoka, Hiroyuki; Sakiyama, Norihisa; Ueno, Yukizo; Nakai, Kiyohiko; Makita, Masami

doi:10.1385/1-59259-047-0:183

Cited by 1 publication

(1 citation statement)

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“…A number of analytical techniques have been exploited for phosphoamino acids analysis, as found in two review articles 4, 5, such as radioactive labeling 6, antibody recognition 7, HPLC 3, 8–10, GC 11, 12, CE 13–20, and MS 21. In comparison to other methods, CE represents a sensitive and high‐resolution separation method that is well suited for the analysis of trace levels of phosphoamino acids in biological samples due to its rapid analysis times, minimal band dispersion, and small sample requirements.…”

Section: Introductionmentioning

confidence: 99%

Rapid analysis of phosphoamino acids from phosvitin by near‐infrared cyanine 1‐(ε‐succinimydyl‐hexanoate)‐1′‐methyl‐3,3,3′,3′‐tetramethyl‐indocarbocyanine‐5,5′‐disulfonate potassium derivatization and polyacrylamide‐coated CE with LIF detection

Wang

Zhang

2011

Electrophoresis

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A polyacrylamide-coated CE method with LIF detection was developed for analyzing three phosphoamino acids including phosphotyrosine (p-Tyr), phosphothreonine (p-Thr), and phosphoserine (p-Ser). A near-infrared dye, 1-(ε-succinimydyl-hexanoate)-1'-methyl-3,3,3',3'-tetramethyl-indocarbocyanine-5,5'-disulfonate potassium (MeCy5-OSu) was employed for derivatization of these phosphoamino acids. Results indicated that the complete baseline resolution of each phosphoamino acid was obtained within 6.1 min, using 10 mmol/L phosphate buffer (pH 4.0) containing 60 mmol/L SDS as running buffer. The highest derivatization efficiency was achieved in 0.2 mol/L borate buffer (pH 8.8) for 30 min at 30 °C. Linearity of response was found in the range of 0.05-1 μmol/L. The correlation coefficients for these phosphoamino acids were from 0.9940 to 0.9976. The LODs for phosphotyrosine, phosphothreonine, and phosphoserine were about 6, 8, and 8 nmol/L, respectively. The proposed method has been successfully applied to the determination of phosphoamino acids in the hydrolysis sample of a phosphorylated phosvitin. Average recoveries for phosvitin sample were in the range of 94.0-98.0% and coefficients of variation ranged from 2.7 to 4.8%.

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