巴旦木青皮化学成分分析及生物活性评价

发布时间：2018-05-16 23:28

本文选题：巴旦木青皮 + 黄酮　；参考：《新疆大学》2016年硕士论文

【摘要】：新疆巴旦木(Amygdalus communis L.)资源丰富,但目前对巴旦木的利用仅仅局限于果仁及其粗加工品,其中占巴旦木果实20~30%的青皮作为废弃物没有被充分利用。因此,通过对巴旦木青皮中的黄酮类物质及三萜类物质等有效活性成分的研究,为巴旦木资源的综合利用提供理论基础和技术支持。本文用巴旦木青皮为研究试材,就巴旦木青皮中有效化学成分的初步提取与分离、活性物质含量的测定以及提取物的抗氧化与抑菌能力的评价展开研究。主要结论如下:1、对巴旦木青皮(新疆莎车县)中常规化学成分进行分析,测定出水分、灰分、粗脂肪、氨基酸、总糖、总多酚和总三萜含量分别为3.71±0.09%、13.30±0.18%、1.85±0.05%、2.08%、4.18±0.07%、0.59±0.01%和1.41±0.03%。2、采用超声波辅助有机溶剂提取法对巴旦木青皮中总黄酮进行提取,优化后的提取条件为:提取剂100%甲醇,超声时间30 min,料液比1:30(g/mL),提取次数3次,在该条件下总黄酮提取量可达31.94±0.36 mg/g,RSD为1.13%(n=3)。3、建立了分离巴旦木青皮中芦丁(Rut)、金丝桃苷(Hyp)和紫云英苷(Ast)的高效液相色谱(HPLC)分析方法。结果表明:Rut标准品的质量浓度和峰面积在0.1~100.0μg/mL的浓度范围内线性关系良好(R2=0.9999),回收率为93.24%~99.95%,RSD为1.92%~3.37%(n=3);Hyp标准品的质量浓度和峰面积在0.1~100.0μg/mL浓度范围内线性关系良好(R2=0.9998),回收率为94.95%~101.54%,RSD为1.17%~2.95%(n=3);Ast标准品的质量浓度和峰面积在0.1~100.0μg/mL的浓度范围内线性关系良好(R2=0.9999),回收率为92.37%~99.58%,RSD为1.24~3.41%(n=3);并采用高分辨质谱法(HRMS)对实验结果进一步进行了验证。4、建立了分离巴旦木青皮中齐墩果酸(OA)和熊果酸(UA)的HPLC分析方法。结果表明,OA标准品的质量浓度和峰面积在5~500μg/mL的浓度范围内线性关系良好(R2=0.9999),回收率为89.67%~94.85%,RSD为1.28%~3.16%(n=3);UA标准品的质量浓度和峰面积在5~500μg/m L的浓度范围内线性关系良好(R2=0.9999),回收率为88.67%~94.77%,RSD为1.07%~1.92%(n=3);并采用HRMS对实验结果进一步进行了验证。5、采用超声波辅助有机溶剂提取法对巴旦木青皮中的OA和UA进行提取,优化后的提取条件为:提取剂为95%乙醇、料液比为1:40(g/mL)、提取时间为40 min及提取次数为3次。在该提取条件下,OA的提取量可达1.01±0.003mg/g,RSD为2.97%(n=3),UA提取量可达0.68±0.002 mg/g,RSD为2.94%(n=3)。6、采用1,1-二苯基-2-三硝基苯肼(DPPH)自由基、羟基(OH)自由基、2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二胺盐(ABTS)自由基清除法以及还原力测定法等四种指标考察了巴旦木青皮甲醇提取物和乙醇提取物体外抗氧化能力的强弱。结果表明,提取物均具有一定的抗氧化能力,但与阳性对照物芦丁(Rutin)、维生素C(Vitamin C)、2,6-二叔丁基-4-甲基苯(BHT)相比,抗氧化能力低于对照组,可能是由于所含抗氧化物质的纯度和化学结构的不同导致。7、采用琼脂扩散法和生长速率法评价了巴旦木青皮甲醇提取物和乙醇提取物对金黄色葡萄球菌、枯草芽孢杆菌以及镰刀菌、大丽轮枝菌的抑制作用。结果表明,提取物对所选菌种具有一定的抑制作用。
[Abstract]:The Amygdalus communis L. (Amygdalus communis) is rich in resources, but at present, the use of pidanwood is limited to kernel and its crude products, and the peel of 20~30% from the fruit of the Panna is not fully used as waste. This paper provides a theoretical basis and technical support for the comprehensive utilization of the resources of the pericarwood. This paper studies the preliminary extraction and separation of the effective chemical components in the pericarpine green skin, the determination of the content of active substances and the evaluation of the antioxidative and bacteriostasis ability of the extract. The main conclusions are as follows: 1. The chemical composition of skin (Xinjiang Shache county) was analyzed. The content of water, ash, crude fat, amino acid, total sugar, total polyphenols and total three terpenes were 3.71 + 0.09%, 13.30 + 0.18%, 1.85 + 0.05%, 2.08%, 4.18 + 0.07%, 0.59 + 0.01% and 1.41 + 0.03%.2, respectively. The extraction conditions were as follows: 100% methanol extraction agent, 30 min ultrasonic time and 3 times of extract ratio at 1:30 (g/mL). The extraction amount of total flavonoids can reach 31.94 + 0.36 mg/g and RSD is 1.13% (n=3).3. The high performance liquid chromatography (HPLC) for separation of rutin (Rut), Hyperoside (Hyp) and cloud oside (Ast) in the peel of pericarwood is established. The results showed that the linear relationship between the mass concentration and peak area of Rut standard was good (R2=0.9999) in the concentration range of 0.1~100.0 g/mL, the recovery rate was 93.24%~99.95%, RSD was 1.92%~3.37% (n=3), and the quality concentration and peak area of Hyp standard products were well (R2=0.9998) in 0.1~100.0 um g/mL concentration range (R2=0.9998), and the recovery rate was 94.95. %~101.54%, RSD is 1.17%~2.95% (n=3); the mass concentration and peak area of Ast standard products have a good linear relationship with the concentration range of 0.1~100.0 mu g/mL (R2=0.9999), the recovery rate is 92.37%~99.58%, RSD is 1.24~3.41% (n=3), and the experimental results are further verified by high resolution mass spectrometry. The results of HPLC analysis of oleic acid (OA) and ursolic acid (UA) show that the mass concentration and peak area of OA standard products have a good linear relationship in the concentration range of 5~500 / g/mL (R2=0.9999), the recovery rate is 89.67%~94.85%, and RSD is 1.28%~3.16% (n=3), and the mass concentration and peak area of the UA standard products have a good linear relationship with the concentration range. (R2=0.9999), the recovery rate is 88.67%~94.77%, RSD is 1.07%~1.92% (n=3), and HRMS is used to further verify the experimental results.5, and the ultrasonic assisted organic solvent extraction is used to extract OA and UA in the pericarpine green skin. The optimum extraction conditions are as follows: the extraction agent is 95% ethanol, the ratio of material to liquid is 1:40 (g/mL), and the extraction time is 40 min. And the extraction times are 3 times. Under this extraction condition, the extraction of OA can reach 1.01 + 0.003mg/g, RSD is 2.97% (n=3), the extraction of UA is 0.68 + 0.002 mg/g, RSD is 2.94% (n=3).6, 1,1- two phenyl -2- three nitrophenyl hydrazine (DPPH) radical, hydroxyl radical and two amine salt free radical, two (ethyl benzothiazole sulfonic acid) two amine salt free radical Four indexes, such as division and reductive force determination, were used to investigate the antioxidant capacity of the extractive and ethanol extracts from the carbinol and ethanol extracts of pericarwood. The results showed that the extracts all had a certain antioxidant capacity, but compared with the positive control rutin (Rutin), vitamin C (Vitamin C), 2,6- ditert butyl -4- methyl benzene (BHT), antioxidant energy. The force is lower than the control group, which may be due to the purity of the antioxidants and the difference in the chemical structure that lead to.7. The inhibition effect of the methanol extract and ethanol extract from the peel of the pericarp on Staphylococcus aureus, Bacillus subtilis, Fusarium, Fusarium and dacydendron is evaluated by the agar diffusion method and the growth rate method. The extract has a certain inhibitory effect on the selected strains.
【学位授予单位】：新疆大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R29

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