Kazarjan Jana, Mahlapuu Riina, Hansen Mats, Soomets Ursel, Kaljurand Mihkel, Vaher Merike
Department of Chemistry, Tallinn University of Technology, Tallinn, Estonia.
Institute of Biomedicine and Translational Medicine, Department of Biochemistry, Centre of Excellence for Translational Medicine, University of Tartu, Tartu, Estonia.
J Sep Sci. 2015 Oct;38(19):3461-8. doi: 10.1002/jssc.201500567. Epub 2015 Aug 21.
In the present study, a micellar electrokinetic chromatographic method was used to determine the retention factors of hydrophilic monomeric and homodimeric forms of glutathione analogues. Ionic-liquid-based surfactant, 1-tetradecyl-3-methylimidazolium chloride, as well as cetyltrimethylammonium bromide and phosphate buffer (pH 7.4) were employed in the experiments. Since the studied peptides possess a negative charge under physiological conditions, it is expected that the peptides interact with the oppositely charged 1-tetradecyl-3-methylimidazolium chloride and cetyltrimethylammonium bromide micelles via hydrophobically assisted electrostatic forces. The dependence of the retention factor on the micellar concentration of 1-tetradecyl-3-methylimidazolium chloride and cetyltrimethylammonium bromide is nonlinear and the obtained curves converge to a limiting value. The retention factor values of GSH analogues were in the range of 0.36-2.22 for glutathione analogues and -1.21 to 0.37 for glutathione when 1-tetradecyl-3-methylimidazolium chloride was used. When cetyltrimethylammonium bromide was employed, the retention factor values were in the range of 0.27-2.17 for glutathione analogues and -1.22 to 0.06 for glutathione. If sodium dodecyl sulfate was used, the retention factor values of glutathione analogues with carnosine moiety were in the range of -1.54 to 0.38.
在本研究中,采用胶束电动色谱法测定谷胱甘肽类似物的亲水性单体和同二聚体形式的保留因子。实验中使用了离子液体基表面活性剂1-十四烷基-3-甲基咪唑氯盐,以及十六烷基三甲基溴化铵和磷酸盐缓冲液(pH 7.4)。由于所研究的肽在生理条件下带负电荷,预计这些肽会通过疏水辅助静电力与带相反电荷的1-十四烷基-3-甲基咪唑氯盐和十六烷基三甲基溴化铵胶束相互作用。保留因子对1-十四烷基-3-甲基咪唑氯盐和十六烷基三甲基溴化铵胶束浓度的依赖性是非线性的,所得曲线收敛到一个极限值。当使用1-十四烷基-3-甲基咪唑氯盐时,谷胱甘肽类似物的保留因子值在0.36 - 2.22范围内,谷胱甘肽的保留因子值在 - 1.21至0.37范围内。当使用十六烷基三甲基溴化铵时,谷胱甘肽类似物的保留因子值在0.27 - 2.17范围内,谷胱甘肽的保留因子值在 - 1.22至0.06范围内。如果使用十二烷基硫酸钠,带有肌肽部分的谷胱甘肽类似物的保留因子值在 - 1.54至0.38范围内。