采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、双抗夹心酶联免疫吸附(enzyme-linked immunosorbent assay,ELISA)等方法,以Jug r 1含量为指标,对云南7个不同产地的泡核桃(Juglans sigillata)进行筛选,进一步利用硫酸铵分级沉淀、凝胶过滤层...采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、双抗夹心酶联免疫吸附(enzyme-linked immunosorbent assay,ELISA)等方法,以Jug r 1含量为指标,对云南7个不同产地的泡核桃(Juglans sigillata)进行筛选,进一步利用硫酸铵分级沉淀、凝胶过滤层析、液相色谱-串联质谱等手段制备Jug r 1并进行优化和鉴定,通过圆二色谱对Jug r 1的结构进行表征,最后采用双抗夹心ELISA法测定其含量。结果表明:保山隆阳产地核桃的总蛋白含量和Jug r 1含量均最高,用于后续实验;经优化得到Jug r 1的最佳硫酸铵分级沉淀分离区间为40%~80%,进一步对其进行凝胶过滤层析的最佳条件为上样质量浓度30 mg/mL、上样体积4 mL、洗脱流速1 mL/min;在此条件下纯化得到Jug r 1含量达19.90 mg/120 mg上样量,蛋白得率为16.58%;质谱分析表明该蛋白符合Jug r 1的典型特征;圆二色谱显示Jug r 1的二级结构以α-螺旋为主,多种构象共存;经“两步法”分离纯化后可获得蛋白纯度占总蛋白96%以上的Jug r 1。本结果可为深入研究Jug r 1提供科学基础,并为其他坚果致敏蛋白的分离纯化提供参考。展开更多
Separation of shelled walnut particles was studied on two varieties of Persian walnut,Poost-Kaghazi and Poost-Sangi using pneumatic method.The moisture contents of the samples were determined.The particles were consid...Separation of shelled walnut particles was studied on two varieties of Persian walnut,Poost-Kaghazi and Poost-Sangi using pneumatic method.The moisture contents of the samples were determined.The particles were considered in three categories of shell,kernel and shell-kernel together.Each category was manually classified based on their size,in three portions of 1/8,1/4,and 1/2,as well as the whole kernel and whole walnut.The terminal velocity of each group was determined.The shelled walnuts were sieved and classified in three groups of small,medium and large.The effects of separation time(5,10 and 15 seconds),feeding value(50 to 80 gr)and air velocity on separation of the kernels and shells were studied for both varieties.The interaction effects were also studied for three walnut sizes(small,medium and large).The terminal velocity was the highest for the whole walnut and the whole kernel while it was lowest for 1/4 and 1/8 of the shell.The best separation was performed at air velocities of 9.20,10.04 and 10.94 m/s with 98.2%,98.9%and 98.2%,respectively.展开更多
通过吸附、解吸试验,筛选适合分离纯化核桃青皮多酚的大孔树脂,并对分离纯化条件进行优化。以没食子酸、原儿茶素等9种标准品绘制标准曲线,采用HPLC对所得核桃青皮多酚物质进行定性和定量分析。结果表明:NKA-9树脂是分离纯化核桃青皮多...通过吸附、解吸试验,筛选适合分离纯化核桃青皮多酚的大孔树脂,并对分离纯化条件进行优化。以没食子酸、原儿茶素等9种标准品绘制标准曲线,采用HPLC对所得核桃青皮多酚物质进行定性和定量分析。结果表明:NKA-9树脂是分离纯化核桃青皮多酚的最佳树脂,吸附条件为上样液多酚浓度0.574 mg/m L,p H 1.0,上样速率1.3 m L/min,吸附率可达90.1%;洗脱条件为体积分数50%的乙醇洗脱剂,p H 3.0,洗脱速率0.43 m L/min,解吸率达87.9%。经HPLC鉴定和分析,核桃青皮酚类物质主要由原儿茶素、绿原酸、咖啡酸、表儿茶素、阿魏酸、芦丁等单体组成,其中含量较高的为表儿茶素、原儿茶素和芦丁。展开更多
比较6种不同极性大孔树脂对核桃青皮提取物中胡桃醌的静态吸附和解吸性能,筛选出最佳的大孔树脂。通过与胡桃醌标准品HPLC保留时间相比较的方法对胡桃醌含量进行分析。结果表明:D101树脂为纯化核桃青皮胡桃醌的最佳树脂,较佳的动态吸附...比较6种不同极性大孔树脂对核桃青皮提取物中胡桃醌的静态吸附和解吸性能,筛选出最佳的大孔树脂。通过与胡桃醌标准品HPLC保留时间相比较的方法对胡桃醌含量进行分析。结果表明:D101树脂为纯化核桃青皮胡桃醌的最佳树脂,较佳的动态吸附条件为上样液胡桃醌浓度0.114 mg/m L,p H 2.0,上样速率为1.0 m L/min,吸附率可达81.85%;较佳的洗脱条件为体积分数95%的丙酮,p H 4.0,洗脱速率为0.33 m L/min,解吸率为76.5%。经HPLC鉴定和分析,D101分离纯化核桃青皮胡桃醌不会造成组成变化和较大损失。D101大孔树脂对核桃青皮胡桃醌有较好的分离纯化效果。展开更多
文摘采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、双抗夹心酶联免疫吸附(enzyme-linked immunosorbent assay,ELISA)等方法,以Jug r 1含量为指标,对云南7个不同产地的泡核桃(Juglans sigillata)进行筛选,进一步利用硫酸铵分级沉淀、凝胶过滤层析、液相色谱-串联质谱等手段制备Jug r 1并进行优化和鉴定,通过圆二色谱对Jug r 1的结构进行表征,最后采用双抗夹心ELISA法测定其含量。结果表明:保山隆阳产地核桃的总蛋白含量和Jug r 1含量均最高,用于后续实验;经优化得到Jug r 1的最佳硫酸铵分级沉淀分离区间为40%~80%,进一步对其进行凝胶过滤层析的最佳条件为上样质量浓度30 mg/mL、上样体积4 mL、洗脱流速1 mL/min;在此条件下纯化得到Jug r 1含量达19.90 mg/120 mg上样量,蛋白得率为16.58%;质谱分析表明该蛋白符合Jug r 1的典型特征;圆二色谱显示Jug r 1的二级结构以α-螺旋为主,多种构象共存;经“两步法”分离纯化后可获得蛋白纯度占总蛋白96%以上的Jug r 1。本结果可为深入研究Jug r 1提供科学基础,并为其他坚果致敏蛋白的分离纯化提供参考。
文摘Separation of shelled walnut particles was studied on two varieties of Persian walnut,Poost-Kaghazi and Poost-Sangi using pneumatic method.The moisture contents of the samples were determined.The particles were considered in three categories of shell,kernel and shell-kernel together.Each category was manually classified based on their size,in three portions of 1/8,1/4,and 1/2,as well as the whole kernel and whole walnut.The terminal velocity of each group was determined.The shelled walnuts were sieved and classified in three groups of small,medium and large.The effects of separation time(5,10 and 15 seconds),feeding value(50 to 80 gr)and air velocity on separation of the kernels and shells were studied for both varieties.The interaction effects were also studied for three walnut sizes(small,medium and large).The terminal velocity was the highest for the whole walnut and the whole kernel while it was lowest for 1/4 and 1/8 of the shell.The best separation was performed at air velocities of 9.20,10.04 and 10.94 m/s with 98.2%,98.9%and 98.2%,respectively.
文摘通过吸附、解吸试验,筛选适合分离纯化核桃青皮多酚的大孔树脂,并对分离纯化条件进行优化。以没食子酸、原儿茶素等9种标准品绘制标准曲线,采用HPLC对所得核桃青皮多酚物质进行定性和定量分析。结果表明:NKA-9树脂是分离纯化核桃青皮多酚的最佳树脂,吸附条件为上样液多酚浓度0.574 mg/m L,p H 1.0,上样速率1.3 m L/min,吸附率可达90.1%;洗脱条件为体积分数50%的乙醇洗脱剂,p H 3.0,洗脱速率0.43 m L/min,解吸率达87.9%。经HPLC鉴定和分析,核桃青皮酚类物质主要由原儿茶素、绿原酸、咖啡酸、表儿茶素、阿魏酸、芦丁等单体组成,其中含量较高的为表儿茶素、原儿茶素和芦丁。
文摘比较6种不同极性大孔树脂对核桃青皮提取物中胡桃醌的静态吸附和解吸性能,筛选出最佳的大孔树脂。通过与胡桃醌标准品HPLC保留时间相比较的方法对胡桃醌含量进行分析。结果表明:D101树脂为纯化核桃青皮胡桃醌的最佳树脂,较佳的动态吸附条件为上样液胡桃醌浓度0.114 mg/m L,p H 2.0,上样速率为1.0 m L/min,吸附率可达81.85%;较佳的洗脱条件为体积分数95%的丙酮,p H 4.0,洗脱速率为0.33 m L/min,解吸率为76.5%。经HPLC鉴定和分析,D101分离纯化核桃青皮胡桃醌不会造成组成变化和较大损失。D101大孔树脂对核桃青皮胡桃醌有较好的分离纯化效果。
