高液相色谱在研究人工核酸酶中的应用

Application of HPLC in Study of Synthetic Nucleases

目的:用反相高效液相色谱梯度洗脱法测定了用Cu（Ⅱ）组氨酸配合物切割DNA-二核苷酸2’-脱氧腺嘌吟（3’→5’）-2’-脱氧腺嘌呤核苷酸的产物。方法:色谱柱Hypersil BDS C18 150mm×4.6mm;流动相为C:0.2mol·L-1磷酸二氢铵溶液（pH5.5）,B:甲醇-水（3:2）;流速:0.85mL·min-1;梯度为B:0 0→5min;B:0→50％ 5→17min;B,50％→100％17→18min;B:100％19min 停止。紫外检测波长260nm。结果:在本文中,用HPLC检测可得到直接有效的数据。结论:本方法准确、灵敏。

Objective: A RP - HPLC method with gradient elution has been studied to determine the cleavage products of the dinucleotide by Cu (Ⅱ) complex. Methods: Hypersil BDS C18 150 mm ×4.6 mm column; mobile phase, C: 0.2 mol·L-1NH4H2PO4 (PH5.5), B: CH3OH - H2O (3:2); flow rate, 0.85 mL·min-1; gradient, B:00→5 min; B: 0→50% 5→17 min; B: 50%→100% 17→18 min; B:100%19min. Detected at 260 nm. Result: In our report, direct, available data were gained by HPLC . Conclusion: This method is accurate and sensitive.