HPLC检测肠灌流模型中PMP衍生化葡寡糖含量的方法学建立

To evaluate the concentration of 1-Phenyl-3-methyl-5-pyrazolone Derivatized Oligosaccharides in rat intestinal perfusion model using HPLC

目的:建立肠灌流模型中1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生化葡寡糖含量的检测方法,并进行方法学考察。方法:采用HPLC法测定,色谱条件为,色谱柱:phenomenexC18(250×4.60mm,5μm);流动相:100mMNaAc/乙腈(梯度洗脱);流速:1.0mL·min-1;紫外检测波长245nm;进样量:20μL;柱温:40℃。结果:PMP衍生化葡寡糖在此色谱条件下获得良好分离,肠灌流液、血浆样品及酚红对10种化合物的检测均无影响。结论:该方法准确可靠,重复性良好,为寡糖的吸收研究提供了技术支持。

Objective：To establish the method to evaluate the concentrion of PMP derivatized oligosaccharides in rat intestinal perfusion using HPLC and study its methodology.Method：The durg was carried out by resersed-phase technique on a phenomenex C18 column eluted with 100mmol/L ammonium acetate buffer（pH5.0） and acetonitrile as mobile phases in gradient mode.The detection wavelength was 245nm,and the flow rate was 1.0mL·min-1.Result：PMP derivatized oligosaccharides were well separated under the experimental chromatographic conditions,and their detection were not affected by other solutions and reagents.Conclusion：The HPLC method is appropriate for the analysis of PMP derivatized oligosaccharides.This method is simple,quick,accurate,and will provide a technical support to studies on the absorption of oligosaccharide.