Ultra‐performance liquid chromatography coupled to mass spectrometry as a sensitive and powerful technology for metabolomic studies

Abstract

Metabolomics is the comprehensive assessment of endogenous metabolites of a biological system. These large-scale analyses of metabolites are intimately bound to advancements in ultra-performance liquid chromatography-electrospray (UPLC) technologies and have emerged in parallel with the development of novel mass analyzers and hyphenated techniques. Recently, the combination of UPLC with MS covers a number of polar metabolites, thus enlarging the number of detected analytes in the widely used separation sciences. This technology has rapidly been accepted by the analytical community and is being gradually applied to various fields such as metabolomics and traditional Chinese medicine (TCM). Given the power of the technology, metabolomics has become increasingly popular in drug development, molecular medicine, traditional medicine and other biotechnology fields, since it profiles directly the phenotype and changes thereof in contrast to other "-omics" technologies. Hyphenated UPLC/MS technique is becoming a useful tool in the study of body fluids, represents a promising hyphenated microseparation platform in metabolomics and has a strong potential to contribute to disease diagnosis. This review describes the applications of UPLC/MS in metabolomic research, and comparison role of HPLC/MS, NMR and GC/MS, highlights its advantages and limitations with certain characteristic examples in the life and TCM sciences.

Qualitative and Quantitative Analysis of Major Triterpenoids in Alismatis Rhizoma by High Performance Liquid Chromatography/Diode-Array Detector/Quadrupole-Time-of-Flight Mass Spectrometry and Ultra-Performance Liquid Chromatography/Triple Quadrupole Mass Spectrometry

“…The total ion chromatogram in the positive mode of AMR is shown in Figure 2. Based on the structural characteristics of the 25 compounds, they were divided into seven types (as shown in Figure 3): type I includes 16-oxo-11-anhydro-alisol A (6), alisol L (14) and alisol L 23-acetate (20) (19); type VII includes 11-deoxy-alisol C (13) and a new compound named16-oxo-11-deoxy-alisol A (7). All 25 triterpenoids exhibited characteristic fragment ion through dissociation rearrangement between the C-23-C-24; in addition, successive or simultaneous losses of H2O (18 Da) and/ or HAc (60 Da) were observed clearly in their mass spectra, which yielded product Owing to the similar fragmentation pathway of Alisol L, compounds 6 and 20 were identified as 16-oxo-11-anhydro-alisol A and alisol L 23-acetate, respectively, based on UV spectra, retention times and MS data, referring to reference standards and previous reports [13].…”

“…Moreover, triple quadrupole mass spectrometry (QqQ MS) has higher sensitivity than ELSD and UV detections, especially for the non-UV-absorbing such as triterpenoids present in AMR. UPLC coupled with triple quadrupole mass spectrometry (UPLC QqQ MS) with high sensitivity and effectiveness provides a reliable quantification of multi-components in TCM [20,21].…”

Metabolic profiling of human blood

“…По сравнению с газовой хроматографией, ВЭЖХ имеет более низкое хроматографическое разрешение, но это компенсируется более широким диапазоном соединений, которые потенциально могут быть проанализированы. Увеличение разрешающей способности ВЭЖХ достигается за счёт использования более высокой скорости потока, мобильной фазы, увеличения давления и уменьшения диаметра частиц стационарной фазы (менее 2 мкм) (так называемая ультраэффективная жидкостная хроматография (УЭЖХ)) [16]. Применение УЭЖХ разделения на обращенофазовом носителе (С8 или С18) позволяет детектировать масс-спектрометрически до 1000 метаболитов в одном образце [17].…”

Metabolomics: a novel approach to identify potential diagnostic biomarkers and pathogenesis in Alzheimer’s disease

“…(2) to its high throughput, soft ionization, and good coverage of metabolites, as well as its sensitivity and quantitative reproducibility [24] . Similar to HPLC, UPLC allows greater separation and resolution of compounds by decreasing the column size and increasing chromatography pressures [25] .…”