I. Milentyeva, A. Fedorova, T. Larichev, O. Altshuler
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The procedures for phytochemical analysis and isolation of biologically active substances involved such physicochemical research methods as high-performance chromatography (HPLC), thin-layer chromatography (TLC), UV spectrometry, and IR spectrometry. \nThe high performance liquid chromatography confirmed the presence of flavonoids represented by baicalein (5,6,7-trioxyflavone), baicalin (baicalein 7-O-glucuronide), scutellarein (5,6,7,4-tetraoxyflavone), scutellarin (7-O-glucuronide scutellarein), vagonin, and oroxylin. The spectral analyses also detected skutebaicalin. The highest total content of diterpene belonged to the samples extracted with 70% ethanol at 70°C. The content of diterpene was 0.09 mg/cm3 in terms of betulin. The biologically active substances were isolated from the callus extracts of S. baicalensis with a recovery rate of ≥ 80%. 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引用次数: 1
摘要
植物细胞和组织培养物是植物次生代谢物的来源。愈伤组织培养产生的物质可以扩大制药和食品生产的原料基础。然而,从药用植物中分离生物活性物质是一个费时费力的过程。因此,从愈伤组织培养物中提取新的高效技术工艺需求量很大,分离和纯化生物活性物质的新算法仍然是一项相关任务。以黄芩愈伤组织培养为研究对象。植物化学分析和生物活性物质分离的程序涉及到高效色谱法(HPLC)、薄层色谱法(TLC)、紫外光谱法和红外光谱法等理化研究方法。高效液相色谱法证实黄芩苷(5,6,7-三氧黄酮)、黄芩苷(黄芩苷7- o -葡萄糖醛酸盐)、黄芩苷(5,6,7,4-四氧黄酮)、黄芩苷(7- o -葡萄糖醛酸盐黄芩苷)、黄芩苷和芫荽素等黄酮类化合物的存在。光谱分析还检测到黄芩苷。70%乙醇在70℃条件下提取的样品中二萜总含量最高。二萜含量为0.09 mg/cm3(以白介木素计)。从黄芩愈伤组织提取物中分离得到生物活性物质,回收率≥80%。该纯化方案可以获得高纯度的单个生物活性化合物:反式肉桂酸、黄芩苷和oroxylin A的纯度≥95%;黄芩苷纯度≥97%;黄芩素和木犀草素达到≥96%。新工艺提取方法使黄芩愈伤组织的提取物成为可能,并对其成分进行了测定。所建立的分离算法和纯化方案得到的生物活性物质纯化度≥95%。
Biologically active compounds in Scutellaria baicalensis L. callus extract: Phytochemical analysis and isolation
Plant cells and tissue cultures are sources of secondary plant metabolites. Substances produced by callus cultures can expand the raw material base in pharmacy and food production. However, isolating biologically active substances from medicinal plants is a labor- and time-consuming process. As a result, new and efficient technological processes adapted for extraction from callus cultures are in high demand, and new algorithms of isolation and purification of biologically active substances remain a relevant task.
This research featured callus cultures of Scutellaria baicalensis. The procedures for phytochemical analysis and isolation of biologically active substances involved such physicochemical research methods as high-performance chromatography (HPLC), thin-layer chromatography (TLC), UV spectrometry, and IR spectrometry.
The high performance liquid chromatography confirmed the presence of flavonoids represented by baicalein (5,6,7-trioxyflavone), baicalin (baicalein 7-O-glucuronide), scutellarein (5,6,7,4-tetraoxyflavone), scutellarin (7-O-glucuronide scutellarein), vagonin, and oroxylin. The spectral analyses also detected skutebaicalin. The highest total content of diterpene belonged to the samples extracted with 70% ethanol at 70°C. The content of diterpene was 0.09 mg/cm3 in terms of betulin. The biologically active substances were isolated from the callus extracts of S. baicalensis with a recovery rate of ≥ 80%. The purification scheme made it possible to obtain highly-pure individual biologically active compounds: trans-cinnamic acid, baicalin, and oroxylin A had a purity of ≥ 95%; baicalein had a purity of ≥ 97%; scutellarin and luteolin reached ≥ 96%.
The new technological extraction method made it possible to obtain extracts from S. baicalensis callus cultures, which were tested for the component composition. The developed isolation algorithm and purification scheme yielded biologically active substances with a purification degree of ≥ 95%.
期刊介绍:
The journal «Foods and Raw Materials» is published from 2013. It is published in the English and German languages with periodicity of two volumes a year. The main concern of the journal «Foods and Raw Materials» is informing the scientific community on the works by the researchers from Russia and the CIS, strengthening the world position of the science they represent, showing the results of perspective scientific researches in the food industry and related branches. The main tasks of the Journal consist the publication of scientific research results and theoretical and experimental studies, carried out in the Russian and foreign organizations, as well as on the authors'' personal initiative; bringing together different categories of researchers, university and scientific intelligentsia; to create and maintain a common space of scientific communication, bridging the gap between the publications of regional, federal and international level.