高效液相色谱法测定体液及组织样本中D-核糖浓度  

Determination of D-ribose in body fluids and tissue samples by high performance liquid chromatography

作　　者：张泽虹 侯坤 张卿 邰玉 蒲云霞 苏燕[1] ZHANG Ze-hong;HOU Kun;ZHANG Qing;TAI Yu;PU Yun-xia;SU Yan(Baotou Medical College,Baoutou,Inner Mongolia 014000,China;不详)

机构地区：[1]包头医学院,内蒙古自治区包头014000 [2]内蒙古自治区综合疾病预防控制中心,内蒙古自治区呼和浩特010030

出　　处：《中国卫生检验杂志》2022年第15期1793-1795,1815,共4页Chinese Journal of Health Laboratory Technology

基　　金：国家自然科学基金地区基金项目(81860029);内蒙古自然科学基金(2020MS08008);包头医学院2021年研究生科研创新项目(20120202003020103);2021年卫生健康科技计划项目(202201394);2021年卫生健康科技计划项目(202201384)。

摘　　要：目的 改进优化目前尿液及血浆中的D-核糖(D-Ribose)测定的高效液相色谱法(high performance liquid chromatography,HPLC)。方法 在D-核糖-MOPBA衍生法的基础上,通过调整高效液相色谱法流动相的组成、比例以及洗脱时间来优化对D-核糖的分离效果。采用C_(18)色谱柱(4.6 mm×250 mm,5μm),以乙腈为流动相A、离子对试剂为流动相B,洗脱程序为0 min时乙腈溶液占比为10%,10 min时达到15%,18 min上升至100%,20 min~23 min时降至10%。在检测波长为271 nm下对D-核糖进行定性、定量。结果 改进后的D-核糖检测方法保留时间为(15.21±0.20)min,灵敏度更高,能实现目标峰与杂峰的有效分离,可以用于全血、尿液、孵育液及细胞等多种样本中D-核糖浓度的测定。结论 改进后的方法分离效果更好、检测灵敏度更高、适用范围更广,能用于多种体液及组织样本中的D-核糖水平的测定。Objective To improve and optimize the current high performance liquid chromatography(HPLC) method for the determination of D-ribose in urine and plasma.Methods Based on the D-ribose-MOPBA derivatization method,the separation effect of D-ribose-MOPBA was optimized by adjusting the composition,ratio and elution time of the mobile phase.C_(18)column(4.6 mm × 250 mm,5 μm) was used for separation,with acetonitrile as mobile phase A and ion pair reagent as mobile phase B;the proportion of acetonitrile solution is 10% when the elution program is 0 min,reaches 15% at 10 min,rises to100% at 18 min,and decreases down to 10% at 20 min-23 min.The D-ribose was characterized and quantified at the detection wavelength of 271 nm.Results The improved D-ribose detection method has a retention time of(15.21 ± 0.20) min,has high sensitivity and could achieve the effective separation of target and spurious peaks.Besides,it could be used for the determination of D-ribose concentrations in a variety of samples such as whole blood,urine,incubation solution and cells.Conclusion The improved method not only has high sensitivity,but also can achieve effective separation of target and impurity peaks,and also has a wider range of application and is suitable for the determination of D-ribose levels in a variety of body fluids and tissue samples.

关 键 词：D-核糖 高效液相色谱法 血清 尿液 D-核糖-MOPBA 

分 类 号：O657.72[理学—分析化学]