刺玫果提取物化学成分分离及黄酮苷元的制备

发布时间：2018-04-19 23:17

  本文选题：刺玫果 + 黄酮苷元　； 参考：《吉林化工学院》2017年硕士论文

【摘要】：刺玫果为野生山刺玫的成熟果实,分布在东北、内蒙、山西等地。目前广泛地应用在食品添加剂、功能性饮料、保健食品及药品等领域。刺玫果中活性物质丰富,有黄酮、维生素、皂苷、多糖、三萜类等多种化学成分。药理学研究表明,刺玫果具有显著的抗氧化、抗衰老、促消化和抗辐射作用。本文对刺玫果黄酮苷元制备、黄酮苷元含量测定、提取物成分分离等方面进行了探索、研究,进而促进资源丰富、活性广泛的刺玫进一步开发利用。本文首先建立了刺玫果提取物中槲皮素、山奈酚含量的高效液相色谱测定法,并进行了方法学的考察;其次,在单因素考察的基础上,利用响应面法对纤维素酶酶解制备黄酮苷元的工艺条件进行了优化;最后,采用半制备液相、柱层析法、重结晶法等进行刺玫果提取物化学成分的分离,采用薄层色谱法及高效液相色谱法结合法对分离得到的成分进行了结构确证。确立了高效液相色谱同时测定刺玫果提取物中黄酮苷元--槲皮素、山奈酚含量的分析方法。研究结果表明:槲皮素在0.0080~0.6000μg的范围内时,其峰面积与进样量线性关系良好(r=0.9997);山奈酚在0.0025~0.1872μg的范围内时,其峰面积与进样量线性关系良好(r=0.9995)。槲皮素的平均加样回收率为103.92%(RSD=0.29%,n=6);山奈酚的平均加样回收率为95.90%(RSD=0.73%,n=6)。建立的高效液相色谱法操作简便、重现性好、结果准确,可用于刺玫果黄酮苷元的质量控制。以刺玫果提取物为原材料,以酶解后槲皮素峰面积为考察指标,在单因素考察的基础上,采用响应面法优化了酶解法制备刺玫果黄酮苷元的工艺条件。结果表明,当酶解温度为60.2℃、酶解时间为2.1 h、酶解pH值为3.2、酶用量为20.0mg/g、料液比为1:150(g:mL)时,刺玫果黄酮苷元的含量最高。采用生物酶解技术制备黄酮苷元,不但降低了反应温度,还提高了黄酮苷元的得率,具有较大的开发价值。采用半制备液相、柱层析、重结晶法从刺玫果提取物中分离得到四种化合物,通过薄层色谱法及高效液相色谱法对分离成分进行结构确证,四种化合物分别为芦丁、金丝桃苷、槲皮素、山奈酚,为刺玫果提取物的综合利用提供了分子学依据。
[Abstract]:Rosa davidiflora is the mature fruit of wild Rosa davidiana, distributed in Northeast, Inner Mongolia, Shanxi and other places. At present, it is widely used in food additive, functional beverage, health food and medicine. Rosa davidifolia fruit is rich in active substances, including flavonoids, vitamins, saponins, polysaccharides, triterpenoids and other chemical components. Pharmacological studies showed that Rosa davidifolia had significant antioxidant, anti-aging, digestive and radiation effects. In this paper, the preparation of flavone glycosides, the determination of flavonoid glycosides and the separation of extracts from Rosa davidifolia were studied, so as to promote the further development and utilization of Rosa davurica, which is rich in resources and wide in activity. In this paper, a HPLC method for the determination of quercetin and kaempferol in the extract of Rosa davidifolia L. was established, and the methodology was investigated; secondly, on the basis of single factor investigation, the content of quercetin and kaempferol was determined by HPLC. The reaction surface method was used to optimize the process conditions for the preparation of flavonoid glycosides by enzymatic hydrolysis of cellulase. Finally, the chemical constituents of extracts of Rosa davidiflora were separated by semi-prepared liquid phase, column chromatography and recrystallization. The structure of the isolated components was confirmed by TLC and HPLC. A high performance liquid chromatography (HPLC) method was established for the simultaneous determination of flavonoid glycosides quercetin and kaempferol in the extracts of Rosa davurica. The results show that when quercetin is in the range of 0.0080 ~ 0.6000 渭 g, the linear relationship between peak area and sample amount is good, and the peak area of kaempferol in the range of 0.0025 ~ 0.1872 渭 g has a good linear relationship with the sample quantity. The average recoveries of quercetin and kaempferol were 103.92 and 95.90, respectively. The established HPLC method is simple, reproducible and accurate. It can be used for the quality control of flavonoid glycosides in Rosa davidifolia. The extraction of Rosa davurica was used as the raw material and the peak area of quercetin after enzymatic hydrolysis was used as the index. On the basis of single factor investigation, the reaction surface method was used to optimize the process conditions for the preparation of flavonoid glycosides from Rosa davurica by enzymatic hydrolysis. The results showed that when the hydrolysis temperature was 60.2 鈩，

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