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Determination of phosphoamino acids derivatized with 5-(4,6-dichloro-s-triazin-2-ylamino)fluorescein by micellar electrokinetic chromatography.
J Chromatogr A. 2004 Sep 17; 1049(1-2):237-42.JC

Abstract

A sensitive analytical protocol for determining phosphoamino acids using capillary electrophoresis coupled with laser-induced fluorescence detection has been developed. The technique involved the derivatization of the phosphoamino acids with fluorescent reagent 5-(4,6-dichloro-s-triazin-2-ylamino)fluorescein (DTAF) and the analyses of the derivatives by micellar electrokinetic chromatography with laser induced fluorescence detection (MEKC-LIF). Different variables that affect derivatization (DTAF concentration, pH, temperature and time) and separation (kind of surfactant, pH and concentration of buffer) were studied. The baseline separation of three phosphoamino acids could be obtained in less than 11 min with good reproducibility. There was a linear relationship between the peak area of the analyte and its concentration, with correlation coefficients in the range of 0.9979-0.9997. The concentration detection limits (signal to noise = 3) with respect to each single phosphoamino acid were in the range of 0.5-1 nM. The developed method was successfully applied for the determination of phosphoamino acids in the hydrolyzed phosphorylated protein samples.

Authors+Show Affiliations

Liu X
Key Laboratory of MOE for Plant Developmental Biology, College of Life Sciences, Wuhan University, Wuhan 430072, PR China.
Hu YQ
No affiliation info available
Ma L
No affiliation info available
Lu YT
No affiliation info available

MeSH

BuffersCalibrationChromatography, Micellar Electrokinetic CapillaryFluoresceinsFluorescent DyesHydrogen-Ion ConcentrationHydrolysisIndicators and ReagentsPhosphoamino AcidsProtein HydrolysatesProtein KinasesReproducibility of ResultsTobacco

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15499940

Citation

Liu, Xin, et al. "Determination of Phosphoamino Acids Derivatized With 5-(4,6-dichloro-s-triazin-2-ylamino)fluorescein By Micellar Electrokinetic Chromatography." Journal of Chromatography. A, vol. 1049, no. 1-2, 2004, pp. 237-42.
Liu X, Hu YQ, Ma L, et al. Determination of phosphoamino acids derivatized with 5-(4,6-dichloro-s-triazin-2-ylamino)fluorescein by micellar electrokinetic chromatography. J Chromatogr A. 2004;1049(1-2):237-42.
Liu, X., Hu, Y. Q., Ma, L., & Lu, Y. T. (2004). Determination of phosphoamino acids derivatized with 5-(4,6-dichloro-s-triazin-2-ylamino)fluorescein by micellar electrokinetic chromatography. Journal of Chromatography. A, 1049(1-2), 237-42.
Liu X, et al. Determination of Phosphoamino Acids Derivatized With 5-(4,6-dichloro-s-triazin-2-ylamino)fluorescein By Micellar Electrokinetic Chromatography. J Chromatogr A. 2004 Sep 17;1049(1-2):237-42. PubMed PMID: 15499940.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Determination of phosphoamino acids derivatized with 5-(4,6-dichloro-s-triazin-2-ylamino)fluorescein by micellar electrokinetic chromatography. AU - Liu,Xin, AU - Hu,Ye-Qin, AU - Ma,Li, AU - Lu,Ying-Tang, PY - 2004/10/27/pubmed PY - 2005/3/10/medline PY - 2004/10/27/entrez SP - 237 EP - 42 JF - Journal of chromatography. A JO - J Chromatogr A VL - 1049 IS - 1-2 N2 - A sensitive analytical protocol for determining phosphoamino acids using capillary electrophoresis coupled with laser-induced fluorescence detection has been developed. The technique involved the derivatization of the phosphoamino acids with fluorescent reagent 5-(4,6-dichloro-s-triazin-2-ylamino)fluorescein (DTAF) and the analyses of the derivatives by micellar electrokinetic chromatography with laser induced fluorescence detection (MEKC-LIF). Different variables that affect derivatization (DTAF concentration, pH, temperature and time) and separation (kind of surfactant, pH and concentration of buffer) were studied. The baseline separation of three phosphoamino acids could be obtained in less than 11 min with good reproducibility. There was a linear relationship between the peak area of the analyte and its concentration, with correlation coefficients in the range of 0.9979-0.9997. The concentration detection limits (signal to noise = 3) with respect to each single phosphoamino acid were in the range of 0.5-1 nM. The developed method was successfully applied for the determination of phosphoamino acids in the hydrolyzed phosphorylated protein samples. SN - 0021-9673 UR - https://www.unboundmedicine.com/medline/citation/15499940/Determination_of_phosphoamino_acids_derivatized_with_5__46_dichloro_s_triazin_2_ylamino_fluorescein_by_micellar_electrokinetic_chromatography_ DB - PRIME DP - Unbound Medicine ER -
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