Target separation of flavonoids from Saxifraga tangutica using two‐dimensional hydrophilic interaction chromatography/reversed‐phase liquid chromatography

Abstract

An orthogonally (80.3%) preparative two‐dimensional hydrophilic interaction chromatography/reversed‐phase liquid chromatography method has been established for the isolation and purification of flavonoids from Saxifraga tangutica. Initially, flavonoids were enriched by means of a middle‐pressure chromatographic tower (containing middle chromatogram isolated gel). In the first dimension, a XION preparative column was used to separate the flavonoid fractions under the guidance of characteristic ultraviolet absorption spectra of flavonoids and nine flavonoid fractions were obtained. Then, the coeluted flavonoid fractions were selected for further purification via reversed‐phase liquid chromatography with the parent ion peak of quercetin (303), kaempferol (287), or isorhamnetin (317). Several flavonoids could be separated from each hydrophilic interaction chromatography fraction; furthermore, flavonoids with poor resolution in one‐dimensional liquid chromatography were isolated in two‐dimensional liquid chromatography due to the orthogonality. In addition, this technique was valuable for trace flavonoids, which were concentrated in the first stage and separated in the second stage. In total, 18 flavonoids with either quercetin, kaempferol, or isorhamnetin parent nuclei were targetedly obtained, and 15 flavonoids were obtained for the first time from S. tangutica. These results established that the off‐line two‐dimensional hydrophilic interaction chromatography/reversed‐phase liquid chromatography technique was efficient for the isolation of flavonoids from Saxifraga tangutica.

Preparative isolation of 1,1‐diphenyl‐2‐picrylhydrazyl inhibitors from Ribes himalense using medium‐pressure and two‐dimensional reversed‐phase/reversed‐phase liquid chromatography guided by an online HPLC‐1, 1‐diphenyl‐2‐picrylhydrazyl assay

“…Two‐dimensional reversed‐phase/reversed‐phase liquid chromatography (2D RPLC/RPLC) may not be able to effectively separate many sample types owing to the similar separation mechanisms used. However, it is capable of achieving greater peak capacities and degrees of separation than other 2D combinations when eluting at different flow phase pH values or using different organic modifiers [23]. In the present study, a chromatographic column called PTAS was combined with a ReproSil‐Pur C18 AQ chromatographic column to develop a novel 2D RPLC/RPLC purification approach.…”

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

“…Therefore, the NPs including antioxidants that regulate oxidative stress are crucial for preventing or delaying the onset of these diseases. Saxifraga tangutica (S. tangutica) or Song Di, is a famous traditional Tibet Medicine, which is a perennial herb belonging to the Saxifragaceae family [14][15][16]. It is widely distributed at high elevations > 2900 m in Qinghai, Sichuan, and Xizang provinces of West China [17,18].…”

“…It has been used for treating hepatitis, cholecystitis, fever, and trauma. Until now, several compounds, including flavonoids [14], diarylheptanoids [15,18], and phenylpropanoids [15] had been isolated from S. tangutica. However, only eight antioxidants had been isolated from S. tangutica using the fractions activity-oriented strategy [19].…”

Simple and efficient preparation of high‐purity trehalulose from the waste syrup of isomaltulose production using solid‐phase extraction followed by hydrophilic interaction chromatography

“…Though the commercial HILIC TSK Amide-80 and amino column have been reported for the separation of isomaltulose and trehalulose on an analytical scale [17,18], it is difficult to be applied for the purification of trehalulose on a preparative scale due to the low resolution. In 2012, Jin et al developed a silica-based cysteine-bonded zwitterionic HILIC stationary phase (Click TE-Cys) which showed better hydrophilic ability than TSK Amide-80 material [19], and it has been applied for the separation of aminoglycoside antibiotics, anthocyanins, and flavonoids [20][21][22]. However, to the best of our knowledge HILIC has not been described to purify trehalulose from isomaltulose on a semi-preparative scale.…”