酪氨酸对映体的柱前手性衍生高效液相色谱-化学发光联用法研究  

Determination of tyrosine enantiomers by HPLC-CL with precolumn chiral derivatization

作　　者：徐豪杰 高山[1] 任艳丽[1] 荣守华[2] XU Hao-jie;GAO ShanI;REN Yan-li;RONG Shou-hua(Department of Pharmacy,The Third Affiliated Hospital,Zhengzhou University,Zhengzhou 450052,China;Department of Clinieal Laboratory,The Third Affiliated Hospital,Zhengzhou University,Zhengzhou,450052,China)

机构地区：[1]郑州大学第三附属医院药学部,郑州450052 [2]郑州大学第三附属医院检验科,郑州450052

出　　处：《药物分析杂志》2018年第11期1994-1999,共6页Chinese Journal of Pharmaceutical Analysis

基　　金：河南省教育厅基金资助项目(17A350015)

摘　　要：目的:建立酪氨酸对映体的柱前手性衍生高效液相色谱-化学发光联用新方法,并用于体内外酪氨酸对映体的高效检测。方法:利用柱前手性衍生高效液相色谱法对酪氨酸对映体进行分离,联用化学发光检测器对酪氨酸对映体进行检测。考察了色谱流动相配比、流速、柱温以及化学发光条件对酪氨酸对映体分离结果的影响。采用柱前手性衍生高效液相色谱-化学发光联用法对生物样品中的酪氨酸对映体进行分离分析。结果:最佳色谱条件下,流动相为甲醇-0.01 mol·L^(-1)磷酸二氢钾溶液(35∶65,p H 6.0),流速为0.7 mL·min^(-1),柱温为室温;最优化学发光体系为氢氧化钠、鲁米诺和高锰酸钾,其浓度分别为0.15、4.0×10^(-5)和4.0×10^(-5) mol·L^(-1)。在此优化条件下,酪氨酸对映体可以达到完全分离(RS>2.0),线性检测范围为2.5×10^(-7)~3.2×10^(-5) g·m L^(-1),L-/D-酪氨酸的检测下限分别为2.96×10^(-8)、3.59×10^(-8) g·m L^(-1)。结论:柱前手性衍生高效液相色谱-化学发光联用法可对酪氨酸对映体进行高选择性、高灵敏度检测。Objective：To establish a novel HPLC-CL method with precolumn chiral derivatization for the determine of tyrosine enantiomers,and to detect chiral substances in vivo and in vitro.Methods：The tyrosine enantiomers were separated by HPLC with precolumn chiral derivatization and detected by chemiluminescence detector;the chromatographic mobile phase,flow rate,column temperature and chemiluminescence conditions were investigated;the established method was used to analyze the tyrosine enantiomers in biological samples.Results：The chromatographic mobile phase consisted of methanol and 0.01 mol·L-1 monopotassium phosphate solution（35∶65,pH 6.0）,the flow rate was 0.7 mL·min-1,and the column temperature was room temperature.The chemiluminescence system was composed of sodium hydroxide（0.15 mol·L-1）,luminol（4.0×10-5 mol·L-1）and potassium permanganate（4.0×10-5 mol·L-1）.Under the optimized conditions,the tyrosine enantiomers could be completely separated（RS2.0）with the linear range of 2.5×10-7 ~3.2×10-5 g·mL-1,and the detection limits of L-and D-tyrosine were 2.96×10-8 and 3.59×10-8 g·mL-1,respectively.Conclusion：The established method can be used to selectively and sensitively determine tyrosine enantiomers.

关 键 词：柱前衍生化 手性色谱法 化学发光法 酪氨酸对映体 高灵敏 

分 类 号：R917[医药卫生—药物分析学]