摘要
采用高效液相色谱法HPLC对微透析液进行痕量脯氨酸的检测,研究了痕量脯氨酸柱前衍生化的方法。采用苯异硫氰酸酯PITC-乙腈溶液和三乙胺-乙腈溶液对透析液中痕量脯氨酸进行衍生化后,通过环己烷萃取,得到透析液中痕量脯氨酸衍生化的最适方法,无需后续处理可直接进样,并确定了HPLC检测条件:岛津Inertsil ODS-SP色谱柱(250 mm ×4.6 mm,5μm);柱温:31℃;流动相A:0.05 mol·L-1醋酸钠溶液;流动相 B:乙腈-流动相 A(1:1);洗脱条件:A:B(50:50);流速:1.0 mL· min-1;检测波长:254 nm;进样量:20μL。脯氨酸加标回收率为95.1%~104.2%。同时,对3种杨树质外体透析液中的脯氨酸进行了测量。
Based on microdialysis experience in apoplast of poplar stems, the trace of proline in plant was detected by using the High -performance liquid chromatography ( HPLC ) method, precolumn derivatization method was studied. Phenyl isothiocyanate PITC- acetonitrile solution and triethylamine acetonitrile solution were applied to derivate the proline. Cyclohexane was used to extract proline, the sample could be directly injected in HPLC without subsequent pro-cessing. The detection conditions were determined: the SHIMADZU Inertsil ODS-SP column (250 mm × 4. 6 mm, 5 μm);column temperature:31 ℃;mobile phase A:0. 05mol·L-1 so-dium acetate solution, mobile phase B: acetonitrile-mobile phase A (1:1); Elution condi-tions:A:B (50:50);flow rate:1. 0 mL·min-1;detection wavelength:254nm;the injection volume was 20μL. The recovery rate of proline was 95. 1 -104. 2%. At last the proline con-tents of three poplar apoplast dialysis were detected by this method.
出处
《大连民族学院学报》
CAS
2015年第1期27-30,共4页
Journal of Dalian Nationalities University
基金
中央高校基本科研业务费专项资金资助项目(DC12010305
DC12010204)
大连民族学院大学生创新创业训练资助项目(X2013030)
关键词
脯氨酸
微透析
杨树
HPLC
proline
microdialysis
poplar
HPLC