Rapid Isolation of Cyanidin 3-Glucoside and Peonidin 3-Glucoside from Black Rice (Oryza sativa) Using High-Performance Countercurrent Chromatography and Reversed-Phase Column Chromatography

Anthocyanins are water soluble plant pigments which are responsible for the blue, red, pink, violet colors in several plant organs such as flowers, fruits, leaves and roots. In recent years, anthocyanin-rich foods have been favored as dietary supplements and health care products due to diverse biological activities of anthocyanins including antioxidant, anti-allergic, anti-diabetic, anti-microbial, anti-cancer and preventing cardiovascular disease. High-performance countercurrent chromatography (HPCCC) coupled with reversed-phase medium pressure liquid chromatography (RP MPLC) method was applied for the rapid and efficient isolation of cyanidin 3-glucoside (C3G) and peonidin 3-glucoside (P3G) from black rice (Oryza sativa L., Poaceae). The crude black rice extract (500 mg) was subjected to HPCCC using two-phase solvent system composed of tert-butyl methyl ether/n-butanol/acetonitrile/0.01% trifluoroacetic acid (TBME/B/A/0.01% TFA, 1 : 3 : 1 : 5, v/v, flow rate - 4.5 mL/min, reversed phase mode) to give enriched anthocyanin extract (37.4 mg), and enriched anthocyanin extract was sequentially chromatographed on RP-MPLC to yield C3G (16.5 mg) and P3G (8.7 mg). The recovery rate and purity of isolated C3G were 76.0% and 98.2%, respectively, and those of P3G were 58.3% and 96.3%, respectively. The present study indicates that HPCCC coupled with RP-MPLC method is more rapid and efficient than multi-step conventional column chromatography for the separation of anthocyanins.

A New Procedure for Separation and Purification of Scutellarin from Erigeron multiradiatus / 中草药·英文版

Objective: To establish a new procedure for isolating scutellarin from Erigeron multiradiatus. Methods: A proposed method was developed by combining macroporous resins with ODS column. Firstly, E. multiradiatus was extracted by ultrasound with 80% methanol. Preliminary separation was performed on macroporous resin column. The performance and adsorption characteristics of three macroporous resins, D140, D141, and D605, were compared and the enrichment procedure was optimized. Further purification was carried out by medium pressure liquid chromatography (MPLC) with ODS column. Results: It was demonstrated that D141 had better extractive effects on scutellarin. The MPLC conditions were optimized as follows: 15% ethanol aqueous as mobile phase with flow rate at 2.5 mL/min. The yield and purity of the isolated scutellarin were 1.20 mg/g and 96.5%, respectively. Conclusion: The overall procedure is efficient and low-cost, which is considered suitable for the separation and purification of scutellarin from E. multiradiatus. The results provide the scientific basis for developing and using scutellarin in clinic. © 2013 Tianjin Press of Chinese Herbal Medicines.

Separation and preparation of polyphenols from leaves of Olea europaea by preparative medium pressure liquid chromatography / 中草药

Objective: To study the material basis of the anti-diabetic effective fraction in the leaves of Olea europaea. Methods: Preparative medium pressure liquid chromatography (PMPLC) was used for the separation and purification of the compounds. Their structures were elucidated by spectral analyses. Results: The structures of the compounds were identified by various spectroscopic methods combined with the data from some published literatures. There were five compounds which were extracted from the anti-diabetic effective fraction in the leaves of O. europaea and identified as decaffeoylverbascoside (1), luteolin-7-O-β-D-glucoside (2), 6″-O-β-D-glucopyranosyl oleuropein (3), oleuroside (4), and oleuropein (5). Additionally, decaffeoylverbascoside is isolated from this plant for the first time and the purities of the five compounds were all above 90% by HPLC detection. Conclusion: This research provides technological support for the development of new drug based on the leaves of O. europaea.