Abstract
The objective of this work was to provide an economic and practical method for the purification of columbianetin-β-D-glucopyranoside from Angelicae Pubescentis Radix extract. In the static adsorption and desorption, the effects of resin type (D101, HP-20, AB-8, GDX-201, and DA201), contact time (10–360 min), and temperature (298–318 K) were assessed on columbianetin-β-D-glucopyranoside adsorption efficiency in laboratory. GDX-201 resin showed the best adsorption and desorption properties for columbianetin-β-D-glucopyranoside. The kinetic data revealed that the equilibrium time for columbianetin-β-D-glucopyranoside adsorption was achieved within 150 min. Moreover, the adsorption kinetic curve was well in accordance with the pseudo-second-order equation (R2 > 0.99). The rate controlling step of the adsorption process was the intraparticle diffusion. The Langmuir isotherm models (R2 > 0.99) could describe the whole adsorption process, which was exothermic and spontaneous according to the result of thermodynamics tests. In the dynamic adsorption and desorption process, the optimum loading flow (4, 5, and 6 BV/h), ethanol concentration (0–60%), and elution volume (10–230 mL) were optimized. Under optimal conditions of 4 BV/h loading flow, 6.7 BV loading volume, 25% ethanol, and 14 BV elution volume, the content of columbianetin-β-D-glucopyranoside in the product was increased 29.61-fold from 0.45% to 13.32 ± 0.64% with yield of 88.03 ± 2.76% by an experiment of lab-scale enlargement. Then, columbianetin-β-D-glucopyranoside was further purified by PHPLC and its purity was more than 98%. Additionally, the analgesic activity of the columbianetin-β-D-glucopyranoside was assessed by hot plate test. The experimental results showed that columbianetin-β-D-glucopyranoside significantly increased the latency of pain response in mice. This study demonstrated columbianetin-β-D-glucopyranoside could be as a potentially natural analgesic component. It could be summed up that the established method was successfully applied to purifying columbianetin-β-D-glucopyranoside from Angelicae Pubescentis Radix extract.
Highlights
Angelicae Pubescentis Radix (APR), the dried roots of Angelica pubescens Maxim. f. biserrata Shan et Yuan, is mainly distributed in Sichuan and Hubei provinces in China
Columbianetinβ-D-glucopyranoside (CBG) is one of coumarin compounds found in APR which has been proven to have the prevention platelet aggregation and protection against glutamate-induced toxicity [5, 6]
In order to get the best purification condition, both dynamic adsorption and desorption tests were executed with a glass column wetpacked with the GDX-201 resin (10 mm × 400 mm, 1 BV 15 mL). e dynamic adsorption tests were carried out by feeding APR extract solution (477.58 μg/mL) at different flow rates (4, 5, and 6 BV/h), and the optimal adsorption speed was screened on the foundation of the adsorbance of the GDX-201 resin column for CBG. e dynamic desorption tests were carried out by using ultrapure water (5 BV) to wash the GDX-201 resin columns firstly and eluted by using 5%–60% ethanol (5 BV) at a speed of 4 BV/h, respectively, and the optimal ethanol solution concentration was chosen based on desorption ratio of the CBG
Summary
Angelicae Pubescentis Radix (APR), the dried roots of Angelica pubescens Maxim. f. biserrata Shan et Yuan, is mainly distributed in Sichuan and Hubei provinces in China. E heating reflux extraction coupled with silica gel column method was usually used to extract and purify coumarin ingredients from Traditional Chinese Medicine [8, 9]. This method is not suitable for industrial scale-up production due to low recovery, being time-consuming, great waste of organic reagents, and environmental pollution. The enriched fraction was further separated and purified by PHPLC On this basis, a hot plate method was established to evaluate CBG analgesic effect. A hot plate method was established to evaluate CBG analgesic effect. e findings contribute to the MAR selection for CBG or other coumarins
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