RP-LC/ESI-MS和HILIC/ESI-MS相结合的大鼠尿液成分检测方法的建立  

Establishment of RP-LC/ESI-MS Combained with HILIC/ESI-MS Method for Research of Metabonomics of Rat Urine

作　　者：高晓燕[1] 白玉静[1] 

机构地区：[1]北京中医药大学科研实验中心,北京100029

出　　处：《中国实验方剂学杂志》2012年第20期119-121,共3页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81173649/H2817);北京中医药大学自主选题项目(5221010061055)

摘　　要：目的:建立反相液相色谱-电喷雾质谱(RP-LC/ESI-MS)和亲水作用液相色谱-电喷雾质谱(HILIC/ESI-MS)相结合的尿液尽量多化学成分检测数据采集方法。方法:RP-LC最佳色谱条件为Prevail C18色谱柱(4.6 mm×250 mm,5μm),柱温25℃,流动相A 10 mmol.L-1甲酸铵缓冲液(甲酸调pH 4.0),B 0.1%甲酸乙腈,A与B梯度洗脱;HILIC最佳色谱条件为Alltima HP HILIC色谱柱(2.1 mm×150 mm,3μm),柱温25℃,流动相A 10 mmol.L-1甲酸铵缓冲液(甲酸调pH 4.0),B0.1%甲酸乙腈,A与B梯度洗脱;质谱条件为ESI离子源,离子源温度350℃,毛细管电压3.5 kV,雾化压力50.0 psi,干燥气流速11.0 L.min-1,质谱扫描范围m/z 15～2 200,正、负离子检出模式。结果:RP-LC/ESI-MS和HILIC/ESI-MS两种色谱系统同时应用对大鼠尿液中极性不同的内源性物质实现了较为全面的检测。结论:该方法可为代谢组学研究中数据采集的全面性提供参考。Objective： To establish a RP-LC/ESI-MS combined with HILIC/ESI-MS method for the research of metabonomics. Method： For RP-LC/ESI-MS, the assay was conducted on a Prevail C18 （4.6 mm x 250 mm, 5 txm）. The mobile phase A was 10 mmol .L-1 ammonium formate （adjusted to pH 4 with formic acid） in water, B was acetonitrile-formic acid （99.9： 0. 1 ）. A gradient elution mode composed of A and B was used as the mobile phase. For HILIC/ESI-MS, the assay was conducted on a Alhima HP HILIC （2. 1 mm x 150 mm, 3 p,m）. The mobile phase A was 10 mmol L-1 ammonium formate （adjusted to pH 4 with formic acid） in water, B was acetonitrile-formic acid （99.9：0.1 ）. A gradient elution mode composed of A and B was also used as the mobile phase. Electrospray ionization （ESI） in the positive and negative ion mode was used as the ionization source. Nitrogen was used as the nebulizer gas and was maintained at a flow of 11.0 L mln-1 with a nebulizer pressure of 50.0 psi. The fragmentor voltage was set at 275.8 V. Mass spectra were recorded covering m/z 15- 2200. Result： Endogenous compounds with different polarity in rat urine could be detected by using RP-LC/ESI- MS and HILIC/ESI-MS. Conclusion： This method could offer a reference for metabonomics study.

关 键 词：反相液相色谱-电喷雾质谱 亲水作用液相色谱-电喷雾质谱 大鼠尿液 全成分检测 

分 类 号：R284.1[医药卫生—中药学]