Because of the greatly different physicochemical properties of metabolites, comprehensive metabolite profiling analysis has always been a challenging task. Reversed-phase liquid chromatography (RPLC) and hydrophilic interaction liquid chromatography (HILIC) have been used to the analysis of nonpolar metabolites and polar metabolites, respectively. In this work, an alternate HILIC/RPLC-mass spectrometry (MS) approach was developed for the comprehensive and high-throughput analysis of polar and nonpolar metabolites. HILIC and RPLC are respectively performed on two ultra-high performance LC (UHPLC) systems, and coupled to one mass spectrometer to acquire the data. When HILIC gradient elution is running RPLC is in a washing and equilibration state, and vice versa. As a result, the total analysis time was reduced by about one third to 25.4 min. Two hundred and eight representative standards including at least twelve types of commonly met metabolites, SRM 1950 plasma, serum, urine and liver tissue samples were used to test the established alternate HILIC/RPLC-MS method. The results demonstrated that the method possessed high metabolite coverage. The developed method was validated to have good linearity and repeatability. As an example of application, 61 significantly changed metabolites in the colon cancer tissues were defined by this established method.

由于代谢物的理化性质差异很大，因此全面的代谢物谱分析一直是一项艰巨的任务。反相液相色谱（RPLC）和亲水作用液相色谱（HILIC）已分别用于分析非极性代谢物和极性代谢物。在这项工作中，开发了另一种HILIC / RPLC质谱（MS）方法，用于极性和非极性代谢物的全面和高通量分析。HILIC和RPLC分别在两个超高性能LC（UHPLC）系统上执行，并与一个质谱仪耦合以获取数据。运行HILIC梯度洗脱时，RPLC处于洗涤和平衡状态，反之亦然。结果，总分析时间减少了约三分之一，至25.4分钟。包括至少十二种常见代谢物，SRM 1950血浆，血清，尿液和肝组织样品在内的208种代表性标准品用于测试已建立的替代HILIC / RPLC-MS方法。结果表明该方法具有较高的代谢物覆盖率。经验证，所开发的方法具有良好的线性和可重复性。作为应用实例，通过这种既定方法确定了结肠癌组织中61种显着变化的代谢产物。经验证，所开发的方法具有良好的线性和可重复性。作为应用实例，通过这种既定方法确定了结肠癌组织中61种显着变化的代谢产物。经验证，所开发的方法具有良好的线性和可重复性。作为应用实例，通过这种既定方法确定了结肠癌组织中61种显着变化的代谢产物。