【作者】 贾雪峰；

【导师】 钟耕；

【作者基本信息】 西南大学 ， 农产品贮藏与加工工程， 2007， 硕士

【摘要】 为了综合利用苦荞麦资源，本研究对处于盛花期（播种后第58天）苦荞麦叶干燥粉碎物中的黄酮进行了定性定量分析，并利用苦荞麦叶为原料生产苦荞黄酮精粉（tartary buckwheat flavonoids powder，TBFP），对提取、纯化工艺进行了优化，测定了TBFP的总黄酮含量、芦丁含量，研究了TBFP功能性，并采用结晶法制备了纯化芦丁，完善了一个快速准确地测定苦荞麦中芦丁含量的高效液相色谱法。实验结论如下：（1）超声波乙醇回流浸提工艺是苦荞麦叶黄酮提取的理想工艺，最佳条件为：超声波（20kHz）处理40min，料液比1:30、pH=8的60％乙醇溶液70℃浸提120min，浸提率达95.26％。（2）本试验中苦荞麦叶黄酮的纯化工艺采用了DA—201小型大孔吸附树脂柱，最佳工艺是：将浸提液真空（60℃，0.09MPa）浓缩；200mL丙酮（AR级）萃取浓缩液，除去杂质；上样液中黄酮浓度为0.5mg／mL以1mL／min通过DA—201树脂柱（f20mm×300mm）；150 mL80％的乙醇（AR级）以1 mL／min洗脱并收集洗脱液；真空（60℃，0.09MPa）浓缩，制得苦荞麦叶黄酮精粉（TBFP）。（3）用苦荞麦叶黄酮精粉（TBFP）进一步结晶精制芦丁工艺：用甲醇加热回流溶解TBFP，过滤，滤液放冷，结晶，过滤得湿粗芦丁。母液回收甲醇，放冷结晶，过滤得湿芦丁，将2次得到的湿芦丁于烘箱中40℃，0.09MPa避光条件烘去大部分甲醇，然后逐渐升温至90℃烘干，然后利用正丁醇加热至80℃左右溶解前步制得的芦丁，过滤，滤液加5mL去离子水，放冷结晶，过滤得湿芦丁，置于0.09MPa避光条件烘箱中90℃烘干得纯芦丁。本工艺制得的纯化芦丁含量和总黄酮含量分别为95.13％、97.89％。（4）HPLC测定苦荞麦中芦丁含量的色谱条件是：色谱柱：Agilent C₁₈，10μm，4.6×250nm不锈钢柱；流动相：甲醇—水—冰乙酸（40:60:1）；流速：1.0mL／min；检测波长：330 nm，柱温：室温（20℃）。芦丁峰形对称，与相邻杂质峰之间能达到基线分离，灵敏度和精密度高，方法简便，芦丁浓度与峰面积显著相关，回归方程为：y=806.16x+297.45，R²=0.999。（5）苦荞麦叶提取物（TBFP）具有清除O₂、清除羟自由基以及还原的能力，且随着黄酮含量的增加而增加，但其清除能力及还原能力要比抗坏血酸的能力弱一些；苦荞麦叶提取物对亚硝酸钠有消除作用，对亚硝酸钠的最大清除率为67.18％。结果表明，此提取物具有较好的阻断清除亚硝酸钠的效果。更多还原

【Abstract】 The leaf which contained the hightest flavonoids among the materials, so the flavonoids from tartary buckwheat leaves were investigated on the character and quantity for the multi-utilization of tartary buckwheat resource in this paper. And tartary buckwheat leaves was selected to produce tartary buckwheat flavonoids powder （TBFP）, and the extraction and purification procedure of TBFP was optimized. A method of extraction and purification of rutin from tartary buckwheat was estabilished. The results were as follow:1. The best technology of extracting flavonoids from tartary buckwheat leaves was as follow: the leaves（5g） was extracted with 150mL, pH=8, 60% ethanol under ultrasonic（20kHz） for 40 minutes, and reflux 120min in a water bath at 70℃.The extraction ratio of flavonoids was 95.26%.2. The column of DA-201 resin was applied in the purification of flavonoids extract, the best technology was as follows:First, condensing the extract to slurry, removing the oiliness by petroleum ether, extracting flavonoids from the slurry for 8h with acetone, filtrating in vacuum, and adapting the content of total flavonoids of extracted solution to 0.5mg/mL.Second, taking 150 mL sample solution （0.5mg/mL） to pass the column at the flow speed of 1.0mL/min.Third, eluting the flavonoids by 150 mL 80% ethanol-water solution at the velocity of 1.0 mL/min.3. Condensing and drying the eluted solution under the vacuum. Refined craft of the buckwheat leaf purification: The methyl alcohol heated the backset current is dissolved, filtrate and put coldly, crystallization. Mother liquor retrieve methyl alcohol, put coldly crystallization, filter, wet 40℃in the oven of rutin 2 times, 0.09MPa photophobic condition bakes most methyl alcohol, then dry to 90℃gradually, the step made rutins to heat and dissolve to about 70℃of then utilized n-butyl alcohol to dissolve, filter, filtrate it with 5mL deionized water, the cold crystallization puts, filter rutins wetly, put 90℃in 0.09MPa photophobic condition oven dry rutins purly. The contents of rutin and total flavonoids of tartary buckwheat flavonoids powder were 95.13%, 97.89%, separately4. The condition of HPLC was: column, Kromasil C18, 10μm, 4.6×250mm stainless steel; mobile phase, methanol—water—acetic acid（40: 60: 1）; flow rate: 1.0 mL/ min; scanning wave-length 330 nm; temperature of column: room temperature（20℃）. The absorbance peak of rutin is symmetry and seperated on the radical line. The conelation of content of rutin and area of peak was y = 806.16x + 297.45 R² = 0.999, the correlation was significant..5. The antioxidizing activity of the extract of shattered leaves was better in scavenging superoxide anion, hydroxyl radical and deoxidizing Fe³⁺. And the capacity of scavenging enhance with the raise of content of total flavonoids in extracting from tartary buckwheat leaves. But the capacity of scavenging of total flavonoids in extracting flavonoids from tartary buckwheat leaves was inferior to Vc. The capability of scavenging sodium nitrite by extract of extracting from tartary buckwheat leaves was 67.18%. The results showed that the extract has high capabilities of scavenging sodium nitrite.更多还原