高效液相色谱中苯甲醛的在线堆积研究及其应用

On-Line Stacking Effect of Benzaldehyde in High Performance Liquid Chromatography and Its Application

以乙腈和水为流动相,研究了苯甲醛在苯基柱和C18柱双柱串联体系中的在线堆积行为。结果表明,苯甲醛在苯基柱-C18柱组合体系中能实现有效堆积,且速度差越大堆积得越好。在实验最佳堆积条件下,堆积因子为4.6,峰高增加5.4倍,该堆积条件下保留时间,峰面积,峰高和半峰宽的RSD值分别为1.2%,4.2%,3.4%和0.80%。将苯甲醛的在线堆积效应用于单胺氧化酶(MAO)酶活性测定样品,信噪比提高了6.9倍,检测灵敏度显著提高。实际样品测定中保留时间,峰面积,峰高,半峰宽,塔板数和信噪比的RSD值分别为0.1%,4.6%,5.0%,0.7%,1.2%和4.67%,精密度满足分析测定的要求。

The on-line stacking effect of benzaldehyde in high performance liquid chromatography （HPLC） based on the velocity difference of the solute on the different stationary phase was discussed by the coupling of two different stationary phases, using H2O and aeetonitrile as mobile phase. The effective stacking was observed and the stacking factor increased with the increase of the velocity difference （the ratio of v1 to v2）. Under the optimal experimental conditions, the stacking factor was 4.6, and the peak height increased 5.4-fold. The RSD of retention time, peak area, peak height and peak width at half peak height was 1.2%, 4.2%,3.4% and 0. 80 % , respectively. Successful stacking was observed by the application of this stacking effect to the real sample in the monoamine oxidase （MAO） activity determination and the signal/noise （S/N） ratio increased 6. 9-fold. In real sample determination, the RSD of retention time, peak area, peak height, peak width at half peak height, plate number and S/N was 0.12%, 4.59%,5.01%,0.70%,1.24% and 4.65%,respectively, which indicated that the stacking effect can be successfully used in the real sample determination with improved sensitivity.