Preparative isolation of antioxidative gallic acid derivatives from Saxifraga tangutica using a class separation method based on medium‐pressure liquid chromatography and reversed‐phase liquid chromatography

Abstract

Saxifraga tangutica is widely used as a medicinal herb to treat hepatic diseases. Here, we developed a class separation method to separate gallic acid derivatives 1,1‐diphenyl‐2‐picrylhydrazyl inhibitors from the methanol extract of Saxifraga tangutica. Firstly, an MCI GEL CHP20P medium‐pressure liquid chromatography was used to pretreat the crude extract from Saxifraga tangutica (500 g) and the target sample (fraction Fr1, 1.7 g) was obtained. Then, an online reversed‐phase liquid chromatography–1,1‐diphenyl‐2‐picrylhydrazyl assay was employed for recognizing potential 1,1‐diphenyl‐2‐picrylhydrazyl inhibitors and six 1,1‐diphenyl‐2‐picrylhydrazyl inhibitors fractions were recognized from fraction Fr1. Subsequently, the six 1,1‐diphenyl‐2‐picrylhydrazyl inhibitors fractions were isolated via a ReproSil‐Pur C18 AQ preparative column. During the separation process, the hydrophilic liquid chromatography was used to enrich the target compounds (Fr1‐3‐1‐1 and Fr1‐3‐1‐2) from the fraction Fr1‐3, which were hardly isolated only by one step reversed‐phase liquid chromatography. Finally, six gallic acid derivatives were obtained and identified as gallic acid (Fr1‐1‐1), gallic acid 3‐O‐β‐D‐glucoside (Fr1‐1‐2), protocatechuic acid (Fr1‐2), 4‐O‐galloyl‐(‐)‐shikimic acid (Fr1‐3‐1‐1), 5‐O‐galloyl‐(‐)‐shikimic acid (Fr1‐3‐1‐2), and 3‐O‐galloyl‐shikimic acid (Fr1‐4), respectively. Thus, the present study indicated that this method was highly efficient for the preparative separation of gallic acid derivatives 1,1‐diphenyl‐2‐picrylhydrazyl inhibitors from natural products.

Screening and Isolation of Potential Anti-Inflammatory Compounds from Saxifraga atrata via Affinity Ultrafiltration-HPLC and Multi-Target Molecular Docking Analyses

“…Instead, target compounds must first be enriched from the crude extracts via sample pretreatment, while non-target components should be eliminated. Based on the principle of analytical liquid chromatography, which takes advantage of the fact that different substances have different partition coefficients in a system consisting of a stationary phase and a mobile phase, medium pressure liquid chromatography (MPLC) represents an effective pretreatment approach [21][22][23]. As such, the preferred approach to the large-scale preparation of high-purity bioactive compounds is a combination of MPLC and prep-HPLC [24,25].…”

Separation Methods of Phenolic Compounds from Plant Extract as Antioxidant Agents Candidate

“…The MPLC system can be applied to adsorption, partition, affinity, or ion-exchange chromatography [80]. The materials used in the stationary phase include silica gel, MCI GEL ® CHP20P, and Sephadex LH-20 [81][82][83]. The choice of column type is critical in MPLC because it can affect the purity and yield.…”

High‐resolution monophenolase/diphenolase/radical scavenging profiling for rapid screening of natural whitening candidates from Rosa rugosa cv. ‘Plena’

“…HPLC combined with microfractionation bioactive screening method is widely employed to identify bioactive components from complex natural products [15]. This method belongs to post‐column principles of biological screening strategies [16].…”