RESUMO
Objective:To study on the material basis of Sanguisorbae Radix by column chromatography and liquid chromatography-ion trap-time-of-flight mass spectrometry (LCMS-IT-TOF), and analyze the distribution of different components in Sanguisorbae Radix water extract on D101 macroporous resin and polyamide resin. Method:Sanguisorbae Radix water extract was separated by D101 macroporous resin and polyamide resin, and LCMS-IT-TOF was used for detection, chromatography separation was achieved on an ACQUITY UPLC HSS T3 column (2.1 mm×100 mm, 1.8 μm) with the mobile phase consisted of water (A) and acetonitrile (B) for gradient elution (0-10 min, 5%-20%B; 10-18 min, 20%-35%B; 18-23 min, 35%-50%B; 23-28 min, 50%-90%B; 28-30 min, 90%B; 30-33 min, 90%-5%B; 33-35 min, 5%B), the flow rate was 0.3 mL·min-1, the column temperature was 30 ℃. Data acquisition was carried out in electrospray ionization (ESI) under the positive and negative ion modes, the scanning range was m/z 100-1 200. According to mass spectrometry data such as accurate molecular mass and fragment information, combined with literature, different chemical components in loading effluents and ethanol eluents of Sanguisorbae Radix water extract were identified. A heat map of the distribution of components in each fraction was drawn by extracting mass spectrum peak intensity data of each sample. The elution rules of various components were compared visually. Result:The enrichment and separation of D101 macroporous resin and polyamide resin were obvious. Tannins in Sanguisorbae Radix water extract was mainly concentrated in loading effluent of macroporous resin and its water eluent, triterpenoids were mainly distributed in the 90% ethanol eluent of macroporous resin. In the above effluents and eluents, a total of 63 compounds (including isomers) were identified. Among them, 6 compounds, ellagic acid-4-pyranoarabinoside or its isomer, 6-O-galloylnorbergerin, 3-O-galloylnorbergerin, (6-acetyloxy-5,7-dihydroxy-8-methoxy-4-oxochromen-2-yl) acetate, ethyl 2-methyl-5,6-bis (sulfooxy) benzofuran-3- carboxylate were first discovered in Sanguisorbae Radix. Conclusion:The method can quickly and accurately identify the distribution of components in aqueous extract of Sanguisorbae Radix after column chromatography, providing experimental basis for exploring the pharmacodynamic components and mechanism of Sanguisorbae Radix.
RESUMO
Objective: Response surface methodology was used to optimize the purification process of naringin from Acanthopanax evodiaefolius leaves by polyamide resin. Methods: The optimum technological conditions for the purification of naringin in the leaves of Acanthopanax evodiaefolius were screened by single factor investigation and response surface design with five factors, including the concentration of sample, sample loading, the elution system, the amount of eluent, and the flow rate. Results: The optimum purification conditions of naringin in the leaves of A. evodiaefolius were as follow: the concentration of the sample was 4.0 mg/mL, the sample volume was 3.5 BV, the elution system was 30% methanol, the eluant volume was 3.0 BV, and the elution flow rate was 8.0 BV/h. Under this condition, the purity of naringin was improved from 5.08% to 56.12%, and the yield was 41.69%. And mass fraction reached more than 90% after recrystallization, which met the requirements of pharmaceutical raw materials. Conclusion: Purification of naringin from the leaves of A. evodiaefolius by polyamide resin chromatography has the advantages of good purification effect, simple operation, high efficiency, and good stability, which can be used for industrial production.