Related Experiment Video
Updated: Apr 13, 2026

Large Scale Non-targeted Metabolomic Profiling of Serum by Ultra Performance Liquid Chromatography-Mass Spectrometry UPLC-MS
Published on: March 14, 2013
An enhanced chemical profiling characterization strategy integrating off-line two-dimensional counter-current
Peiqian Chen1, Yilei Fan2, Xiaoxia Ma1
1College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014, PR China; Zhejiang Provincial Key Laboratory of TCM for Innovative R & D and Digital Intelligent Manufacturing of TCM Great Health Products, PR China; Zhejiang Key Laboratory of Green Manufacturing Technology for Chemical Drug, PR China.
Abstract:
This study establishes a multidimensional analytical platform combining two-dimensional counter-current chromatography (CCC) × ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) with feature-based molecular networking (FBMN) for systematic phytochemical profiling of Bletilla striata fibrous root extract. The orthogonal separation system employed a biphasic solvent system (n-hexane/ethyl acetate/methanol/0.1% formic acid aqueous, 9:11:10:10, v/v) in the first dimension CCC, followed by UHPLC-MS/MS analysis for the secondary separation and characterization. This configuration achieved 80.47% spatial coverage. Total peak capacity was evaluated, and approximately 1328 peaks were obtained through the comprehensive off-line 2D CCC × UHPLC-MS/MS separation. Data-dependent acquisition mode (DDA) generated high-fidelity MS2 spectra, enabled comprehensive structural annotation through visualization, through an integrated workflow combining an in-house database, an online database, and a FBMN analysis. The methodology identified 263 compounds through targeted database matching, with FBMN analysis revealing 15 additional compounds (10 potentially novel natural product and 5 previously unreported in B. striata). Systematic classification identified 142 phenanthrenes, 37 bibenzyls, 16 anthraquinones, and 68 other compounds. Providing a more extensive chemical inventory reported for the fibrous root of B. striata to date (2D CCC × UHPLC-MS/MS 278 total vs. 1D UHPLC-MS/MS 80 total). Advanced visualization tools, including molecular networking and 2D contour plotting, provided more profound insight into structural relationships and chromatographic retention behaviors. This CCC × UHPLC-MS/MS-FBMN platform establishes a new paradigm for comprehensive phytochemical analysis with a broad applicability to natural product research and quality control applications.
More Related Videos
09:04Identifying Per- and Polyfluorinated Chemical Species with a Combined Targeted and Non-Targeted-Screening High-Resolution Mass Spectrometry Workflow
Published on: April 18, 2019
07:50Author Spotlight: Integrating 2D-HPLC-MS and Molecular Networking in Natural Medicine Analysis
Published on: December 8, 2023
Related Concept Videos
Mass Spectrometry: Complex Analysis
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
Peptide Identification Using Tandem Mass Spectrometry
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
Gas Chromatography–Mass Spectrometry (GC–MS)
A gas chromatograph consists of a long, narrow capillary column with a polysiloxane coating on the inner wall....
MALDI-TOF Mass Spectrometry
High-Resolution Mass Spectrometry (HRMS)
Tandem Mass Spectrometry