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KMID : 0043319930160020099
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Archives of Pharmacal Research
1993 Volume.16 No. 2 p.99 ~ p.103
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Improved Phosphotyrosine Analysis by TLC and HPLC
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Song Young-Mi
Yoo Gyurng-Soo
Lee Seung-Ki
Choi Jung-Kap
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Abstract
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We describe here the conditions of thin layer chromatography (TLC) and high pressures liquid chromatography (HPLC) to improve the analytical method of phosphotyrosine (p-Tyr) in biological sample. TLC was performed on silica plate with the mixture of propanol and water (2.1 : 1 v/v) as a mobile phase and values were 0.42, 0.39 and 0.33 for phosphotyrosine, phosphothreonine and phosphoserine, respectively. HPLC was performed on column with a mobile phase of potassium biphosphate solution by UV deterction at 192 nm. The optimum condition of HPLC was obtained at 0.01 M, pH 4.5 with a clear separation within 12 min. These procedures have been applied to the analysis of phosphotyrosine obtained from tyrosine-phosphorylated enolase. Both TLC and HPLC methods were suitable to analyze tyrosine-phosphorylated protein without being affected by contaminants from hydrolysates.
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KEYWORD
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Phosphotyrosine, Phosphoamino acids, Phosphorylated protein, TLC, HPLC
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