Separation of nine novel triterpene saponins from Camellia japonica seeds using high‐performance countercurrent chromatography and reversed‐phase high‐performance liquid chromatography

Abstract

Introduction Camellia japonica L. (Theaceae) is an evergreen shrub, which is cultivated as a popular ornamental tree in Korea, China, and Japan and its seeds have been used as a source of cooking oil, in cosmetics and as a traditional medicine. Intensive phytochemical works have revealed that oleanane‐type saponins are the characteristic compounds of the seeds of C. japonica. Objective The purpose of the present study is to isolate and determine oleanane‐type saponins from C. japonica using high‐performance countercurrent chromatography (HPCCC) coupled with reversed‐phase high‐performance liquid chromatography (RP‐HPLC) and spectroscopic evidences, respectively. Methodology HPLC electrospray ionisation quadrupole time‐of‐flight mass spectrometry (ESI‐Q‐TOF‐MS) was applied to profile the saponin composition of an enriched saponin extract of C. japonica seeds. The enriched saponin extract was separated by HPCCC using a dichloromethane/methanol/isopropanol/water (9:6:1:4, v/v/v/v) system and RP‐HPLC. The structures of the isolates were determined utilising ESI‐Q‐TOF‐MS, one‐dimensional and two‐dimensional NMR and optical rotation. Results HPCCC on enriched saponin extract of C. japonica yielded four saponin fractions in the order of the number of sugars attached to the triterpene aglycone, and preparative RP‐HPLC on each saponin fraction led to the isolation of nine novel saponins, namely camoreoside A–I, along with six known ones. Conclusions This study indicates that combination of HPLC‐ESI‐Q‐TOF‐MS analysis and HPCCC coupled with RP‐HPLC are excellent tools for discovering saponins from natural sources.

Anti-carbapenamase activity of Camellia japonica essential oil against isolated carbapenem resistant klebsiella pneumoniae (MN396685)

“…The exact anti-bacterial compound from the EO was purified by using preparative HPLC using n-Hexane as a active solvent ( Rho et al, 2019 ). After purification, the purified EO material was analyzed by analyticle HPLC for identified the different groups of compounds.…”

Off‐line comprehensive two‐dimensional reversed‐phase countercurrent chromatography with high‐performance liquid chromatography: Orthogonality in separation of Polygonum cuspidatum Sieb. et Zucc

“…The orthogonality of CCC-HPLC was calculated to be 52.23% using Eqs. (1) and (2). The results demonstrated that the combination of RP CCC with HPLC in 2D separations was a powerful approach for the separation of mixtures containing various categories of compounds.…”

“…products including chemical components, peptides and polymers [1][2][3][4][5][6]. It has been widely used for preparative separation of effective components from natural products.…”

“…Combination of CCC and HPLC, also named as 2D chromatography, provides an even more robust and versatile methods for separation of complex matrix, which has been increasingly explored in separation of different natural Article Related Abbreviations: CCC, countercurrent chromatography; 1 D, first dimension; 2 D, second dimension. products including chemical components, peptides and polymers [1][2][3][4][5][6]. On-line comprehensive 2D chromatography has been conventionally applied in gas chromatography (GC × GC) and liquid chromatography (LC×LC) and they have been well studied because of their powerful separation efficiency for complex samples [7,8].…”

Separation of α‐glucosidase inhibitors from Potentilla kleiniana Wight et Arn using solvent and flow‐rate gradient high‐speed counter‐current chromatography target‐guided by ultrafiltration HPLC‐MS screening

“…As a continuous liquid–liquid partition chromatography method, complications of liquid–solid chromatography such as irreversible adsorption, low recovery, and deactivation or loss of activity of sample can be effectively eliminated due to lack of solid support matrix . Thus, HSCCC has been widely used for the separation and purification of active components from natural products, albeit, it is a big challenge to separate all target components directly from crude extract in a single run. Therefore, the stepwise elution mode was introduced into HSCCC separation to address this problem .…”