Comprehensive profiling of Sanguisorba officinalis using offline two‐dimensional mixed‐mode liquid chromatography × reversed‐phase liquid chromatography, tandem high‐resolution mass spectrometry, and molecular network

Abstract

The profiling of natural products is important in modern biological sciences and new drug development. However, the separation and characterization of complex herbal extracts are significantly challenging for researchers in the biochemical field. Herein, an offline two-dimensional mixed-mode liquid chromatography × reversed-phase liquid chromatography system is developed. Our system exhibits high orthogonality and is composed of a newly prepared stationary phase in the first dimension and a traditional C 18 phase in the second dimension, and is operated in combination with a high-resolution mass spectrometry and molecular network. Sanguisorba officinalis L. is studied using the proposed method owing to its bioactivity. With the aid of orthogonal separation, the ionization of the individual components is improved. The number of detected compounds and separated peaks are significantly increased when one-dimensional liquid chromatography is upgraded to two-dimensional liquid chromatography.In addition, 270 compounds (127 of which are tentatively characterized as new compounds, and further confirmation is needed) are successfully characterized based on their fragmentation patterns under the guidance of molecular network, while only 95 compounds are characterized using one-dimensional liquid chromatography and high-resolution mass spectrometry. The results indicate that the developed offline two-dimensional mixed-mode liquid chromatography ×

Sanguisorba officinalis L. promotes diabetic wound healing in rats through inflammation response mediated by macrophage

“…In our recently published work, we developed an off‐line two‐dimensional mixed‐mode liquid chromatography reversed‐phase liquid chromatography coupled with high‐resolution mass spectrometry and a molecular network to exhaustively characterize the ESO. On the basis of their precise molecular mass, fragmentation patterns, and library searches, a total of 270 compounds were tentatively characterized (Dai et al, 2022). In this study, ESO was reanalyzed using the previously described technique, and the fingerprints of ESO and its representative compounds were determined (Figure 2 and Table 2).…”

Rapid identification of the compounds of Bushen Huoxue Prescription based on offline two‐dimensional liquid chromatography with high‐resolution mass spectrometry and molecular network technology

“…When the similarity between two mass spectra is higher than a threshold, they are connected in a MN [22]. This method can infer unknown compounds from known compounds according to the similarity relationship between compounds in the network, thus offsetting the shortcomings of conventional identification methods [11,23]. To improve the quality of an MN, we used Mzmine software to generate a FBMN.…”

“…These works of literature show that 2D-LC with MS improves separation efficiency, reduces ion suppression for subsequent MS detection, and enhances characterization. In addition, there are size exclusion chromatography × RPLC, ion exchange chromatography × RPLC, and other combination modes [11].…”

Pharmacokinetics of four tannin compounds from Sanguisorba officinalis L. before and after processing by ultra‐high‐performance liquid chromatography‐tandem mass spectrometry

“…Modern research indicates that S. officinalis is abundant in chemical components, including tannins, triterpenes, flavonoids, and polysaccharides [8]. Tannins, in particular, represent the highest proportion, accounting for around 17% of the overall composition [9], which can be classified into three main categories: hydrolyzable tannins, condensed tannins, and complex tannins [6].…”