【作者】 付陈梅；

【导师】 阚建全； 刘雄； 陈宗道；

【作者基本信息】 西南农业大学 ， 农产品加工及贮藏工程， 2004， 硕士

【摘要】 本文以重庆江津产青花椒(Z. Schinifolium sieb. et zucc)为原料，利用现代有效成分分离纯化技术、仪器分析鉴定技术对青花椒的呈味物质(花椒麻素)进行了分离、纯化和鉴定；再将纯化得到的晶体作为标样，建立了高效液相色谱法、气相色谱法、紫外分光光度法等定量分析测定方法。结果表明： (1)花椒麻素纯化的工艺流程： 花椒油树脂→麻味物质粗分离→逆流干柱层析→合并2、3部分浸提液→液相色谱柱层析→冷冻结晶→溶解→制备型高效液相色谱纯化→冷冻结晶→结晶体(PC1、PC2、PC3) (2)花椒麻素的理化特征：结晶体均为淡黄色和白色晶体物质，具有强烈的辛麻味；在室温、大气条件下结晶体立即变为黄色粘胶状物质；在充氮、0℃条件下，放置15min，结晶体表面有少量转变为黄色粘胶物质；在-20℃的石油醚溶液中，结晶体稳定。结品体PC1、PC2、PC3的得率分别为0.15％、0.19％、0.09％。 (3)应用薄层色谱检测结晶体纯度，结果发现结晶体(PC1、PC2、PC3)均只有一个色点，其R_f=0.35；应用气-质联谱仪检测，PC2有8个主要色谱峰，其质谱图与花椒麻素的主体物质——不饱和酰胺类物质的质谱图一致。通过鉴定得出，江津青花椒中花椒麻素是由一系列不饱和酰胺类物质组成，主要有8种。结晶体PC2花椒麻素的纯度最高，为81.04％。 (3)以纯度最高的花椒麻素结晶体PC2为标样，建立花椒麻素含量的气相色谱(GC)检测方法。该方法的技术指标：检出限为0.0550μg／mL，回收率范围为87.80％--104.30％，重复性实验中花椒麻素含量的变异系数为1.97％。经多次验证，该方法稳定性强、准确度高，灵敏度高。 (4)以花椒麻素结晶体PC2为标样，建立花椒麻素含量的高效液相色谱(HPLC)检测方法。该方法的技术指标：检出限为0.0440μg／mL，回收率范围为88.10％--104.90％，重复性实验中花椒麻素含量的变异系数为1.57％。目前，该方法已在重庆四面山花椒开发有限公司实际应用了7个月，分析了76批样品，得到652个数据，其准确度和精密度都较好。 (5)以花椒麻素结晶体PC2为标样，建立花椒麻素含量的紫外分光光度检测方法。该方法的技术指标：检出限为0.692μg／mL，回收率范围为81.3％--110.2％，重复性实验测中花椒麻素含量的变异系数为3.71％。经多次验证，该方法的稳定性较好、准确性较高，灵敏度较高，对设备的要求低。 (6)采用所建立的花椒麻素的GC法、HPLC法、及分光光度计法对花椒、花椒调味油以及花椒油树脂中花椒麻素含量进行测定，结果如下：应用GC法、HPLC法、分光光度计法测得花椒中麻素含量分别为15.39mg／g、15.15mg／g、14.71mg／g，测得花椒调味油中麻素含量为1.74mg／g、1.68mg／g、1.85mg／g，花椒油树脂中麻素含量为200.42mg／g、196.65mg／g、西南农业大学硕士学位论文1 88.81m眺。对测定结果进行显著性检验，三种方法测得花椒麻素的均值无显著性差异。 本文为国家高技术研究发展计划(863计划)“高附加值天然调味料及香精制造技术与产品开发(2001AA248021)”和国家2003年度三峡科技移民开发专项“江津花椒产业化及加〕技术开发(2003EP090004)”的部分内容。更多还原

【Abstract】 Used the fruits of Z Schinifolium sieb. et zucc as materials, the numb-taste components were isolated and purified with a series modern technology such as modern isolation and purification technology and instrument analysis technology. Used the purification of numb-taste components as comparison, the quantities of numb-taste components from fruits of Zanthorylum bungemm, oil, and resin were inspected by HPLC, GC and ultraviolet spectrometer. The quantities of water, volatile oil, resin and numb-taste components of fruits with different picking-time were determined. The result indicated that:1. The technology of separating and purifying numb-tasted components:Zanthorylwn bungenum resin →separated with solvent →separated in dry silica gel column with contrary-direction methods → collected the second and third groups →separated in silica gel column eluted with different solvents →collected the peak groups detected by ultraviolet spectrometer →frozen in crystallize in petroleum ether →crystal solved by methyl alcohol →purifled by pre-HPLC →collected the peaks group detected by ultraviolet spectrometer →frozen in crystallize in petroleum ether →crystal (PCl,PC2, PC3)2.The crystals are white or yellow substance which have strong pungent, and the yield rate of PCl, PC2, PC3 reach 0.15%,0.19%, 0.09%. When the crystals are placed in a circumstance with normal temperature and atmosphere, they will turn into yellow viscous substance; when the temperature is 0℃ and nitrogen is filled, only a little of them turn into yellow viscous substance in 15mSnutes; when they are placed in petroleum ether, it is very stable.3. Identified crystal (PCl, PC2, PC3) by thin-layer chromatography, there are only one pot on the thin-layer and the Rf value is 0.35; Identified by HPLC, there are 3 peaks; identified by GC-MS, there are 8 peaks and the mass spectra of crystal are similar to unsaturated alkylamides’. The numb-taste materials of the fruits of Z Schinifolium sieb. et zucc are a mixture of 8 unsaturated alkylamides. The pureness of crystal PC2 is 81.04%, which is the highest of the three.3. Used crystal PC2 as comparison, the GC method was built up to decide the quantities of numb-taste components. The average rate of recovery is 87.8%-104.3%; the coefficient of variation of repeatability test is 1.57%; the limit of detection is 0.055 ug/mL.4. Used crystal PC2 as comparison, the HPLC method was built up to decide the quantities of numb-taste components. The average rate of recovery is 88.1%-104.3%; the coefficient of variation of repeatability test is 1.57%; the limit of detection is 0.044 u g/mL. This method has been used in Chong Qing Si-mian mountain prickly-ash exploit limited company for 7 months, and 76 batches of samples have been analyzed, and 652 data have got, which induced the method is reliable.5. Used crystal PC2 as comparison, the ultraviolet spectrometer method was built up to decide the quantities of numb-taste components. The average rate of recovery is 81.3%-110.2%, the efficient of variation of repeatability test is 3.71%, the limit of detection is 0.692 ug/mL.6. Used GC method, HPLC method and ultraviolet spectrometer method, the contents of numb-taste substances in pericarps of Zanthorylum bungenum are 15.39 mg/g, 15.15 mg/g, and 14.71mg/g, which in oil are 1.74mg/g, 1.68mg/g and 1.85mg/g , which in resin are 200.42mg/g , 196.65mg/g and 188.81 mg/g respectively. By test of significance ,the results are not significant when the significance level is 0.05 .更多还原