番木瓜籽异硫氰酸酯的制备及其特性研究
发布时间：2019-04-09 12:02
【摘要】：异硫氰酸酯(ITCs)是硫代葡萄糖苷(GLS)在植物内源芥子酶作用下的水解产物,主要存在于十字花科芸苔属植物中,作为天然的抗癌、抑菌活性物质备受关注。近年来,在番木瓜籽中也发现含有一定量的硫代葡萄糖苷类物质,本研究以番木瓜籽为原料,研究了苄基硫代葡萄糖苷的酶解工艺,异硫氰酸苄酯的制备工艺,并对提取物进行了包埋。通过气质联用分析推断出番木瓜籽中只含有一种硫代葡萄糖苷,即苄基硫代葡萄糖苷：从结果还可以看出,苄基硫代葡萄糖苷的酶解产物除了异硫氰酸苄酯,还有少量的异氰酸苄酯、苄基腈、硫氰酸苄酯。在单因素试验的基础上进行Box-Behnken试验设计及响应面分析,得出苄基硫代葡萄糖的最佳酶解条件为：原料粒径≥100目,料液比1：20 w/v、酶解缓冲液pH 4.8、酶解时间40 min、酶解温度27℃,此时苄基硫代葡萄糖苷酶解率为94.0%,异硫氰酸苄酯得率为1.35%。通过比较有机溶剂萃取、水蒸汽蒸馏、同时蒸馏萃取三种提取方法,选取同时蒸馏萃取法作为提取方法,提取条件为：二氯甲烷为萃取剂,萃取时间为2 h。粗提物中异硫氰酸苄酯的分离纯化采用硅胶柱层析法,粗提物通过柱层析后纯度由87.8%提高到95.0%。对异硫氰酸苄酯的包埋工艺进行了研究,最佳的包埋工艺为芯材比0.5、包埋温度40℃、包埋时间3 h。通过不同的方法对包埋前后异硫氰酸苄酯的特性进行了研究。相溶解曲线显示,经过包埋后异硫氰酸苄酯的溶解度得到了一定程度的提高；通过热重分析可以看出,包埋后异硫氰酸苄酯的热稳定性得到了提升；X射线衍射图谱显示包埋后β-环糊精的晶体结构发生了明显改变,通过扫描电镜观察到的晶体结构也有明显区别,通过激光光散射仪测得的粒径结果显示,包埋物的粒径比为包埋前环糊精的粒径大,红外光谱和核磁共振结果可用于预测包埋物的分子结构,通过综合分析认为,异硫氰酸苄酯分子结构上的苯环可能部分或者完全进入到环糊精的空腔内,而-N=C=S基团可能被β-环糊精部分包埋。本课题通过对番木瓜籽中苄基硫代葡萄糖苷酶解条件进行研究,实现了异硫氰酸苄酯的提取、分离、纯化和包埋,获得了异硫氰酸苄酯包埋物,为提高番木瓜籽的附加值,研究番木瓜籽中异硫氰酸苄酯的功能性奠定基础。
[Abstract]:Isothiocyanate (ITCs) is the hydrolyzed product of glucosinolate (GLS) under the action of endogenous mustard enzymes in plants. It mainly exists in brassica plants of Cruciferae family. As a natural anticancer substance, its antibacterial activity has attracted much attention. In recent years, a certain amount of glucosinolates was found in papaya seeds. In this study, the enzymatic hydrolysis of benzylthioglucosides and the preparation of benzyl isothiocyanate were studied using papaya seeds as raw materials. And the extract was buried. It was concluded by GC-MS that there was only one glucosinolate in papaya seeds, that is, benzylthioglucosides. The results also showed that the enzymatic hydrolysate of benzylthioglucosides except benzyl isothiocyanate was found to contain only one kind of glucosinolates, that is, benzyl thioglycosides. There are also a small amount of benzyl isocyanate, benzyl nitrile, benzyl thiocyanate. On the basis of single factor test, Box-Behnken test design and response surface analysis were carried out. The optimum conditions for enzymatic hydrolysis of benzylthioglucose were obtained as follows: raw material diameter 鈮，
本文编号：2455164
[Abstract]:Isothiocyanate (ITCs) is the hydrolyzed product of glucosinolate (GLS) under the action of endogenous mustard enzymes in plants. It mainly exists in brassica plants of Cruciferae family. As a natural anticancer substance, its antibacterial activity has attracted much attention. In recent years, a certain amount of glucosinolates was found in papaya seeds. In this study, the enzymatic hydrolysis of benzylthioglucosides and the preparation of benzyl isothiocyanate were studied using papaya seeds as raw materials. And the extract was buried. It was concluded by GC-MS that there was only one glucosinolate in papaya seeds, that is, benzylthioglucosides. The results also showed that the enzymatic hydrolysate of benzylthioglucosides except benzyl isothiocyanate was found to contain only one kind of glucosinolates, that is, benzyl thioglycosides. There are also a small amount of benzyl isocyanate, benzyl nitrile, benzyl thiocyanate. On the basis of single factor test, Box-Behnken test design and response surface analysis were carried out. The optimum conditions for enzymatic hydrolysis of benzylthioglucose were obtained as follows: raw material diameter 鈮，
本文编号：2455164
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