高效液相色谱手性冠醚固定相拆分丙氨酸、正缬氨酸和犬尿氨酸对映体

Enantioseparation of Alanine,Norvaline and Kynurenine by High Performance Liquid Chromatography Using Chiral Crown ether Stationary Phase

采用高效液相色谱法在手性冠醚柱上拆分丙氨酸、正缬氨酸和犬尿氨酸3种对映体。使用CROWN-PAK CR(+)柱(150 mm×4 mm,5μm),以HClO4溶液(pH1.0)为流动相,柱温20℃,流速1.0 mL/min,丙氨酸、正缬氨酸和犬尿氨酸的检测波长分别为200、220和254 nm,3种氨基酸都得到手性拆分。考察了流动相流速、温度及pH对分离的影响,并通过对选择性、保留时间和柱效的考察判断影响分离度的因素。结果表明:降低温度有助于提高手性选择性,但会降低柱效,从而影响分离度。在0～20℃柱温范围内研究了lnk′、lnα与1/T的关系,计算了3种氨基酸对映体与固定相相互作用的热力学函数。

Enantiomers of alanine, norvaline and kynurenine were separated by HPLC using chiral CROWN- PAK CR（ ＋ ） column. Three chiral samples can be separated using pH 1.0 perchloric acid aqueous solution as mobile phase, 20 ℃ as column temperature, and 1.0 mL/min as flow rate. Alanine, norvaline and kynurenine were detected at the wavelengths of 200, 220 and 254 nm, respectively. The effects of flow rate, temperature and pH for separation were investigated. Experimental results show that the lower temperature helps to chiral selectivity, but it also reduces column performance, thereby affectes chiral resolution. The rela- tionship between lnk＇, lnα and 1/T was studied in 0 -20 ℃ o column temperature. The enatioseparation thermodynamic functions of the three enantiomers were calculated.