超声和酶解除杂质法测定烟草中总细胞壁物质含量  

Determination of total cell wall substances content in leaf tobacco by ultrasonic and enzymatic impurity removal method

作　　者：杨蕾[1,2] 陶自伟 潘纯祥 李赛群[2] 李菁菁[2] 郑杰[2] 毕晨曦[2] 侯英[2] 

机构地区：[1]昆明理工大学环境科学与工程学院 [2]云南瑞升烟草技术(集团)有限公司

出　　处：《中国烟草学报》2016年第2期108-114,共7页Acta Tabacaria Sinica

摘　　要：为了减少高毒试剂苯的使用,消除淀粉、蛋白质、灰分对测定总细胞壁物质含量的干扰,本方法采用乙醚-乙醇混合液（乙醚：乙醇=1：15）进行超声脱脂后,加入α-高温淀粉酶,用热水提取,再用中性蛋白酶水解蛋白质,残渣在510℃灰化。结果表明：采用30 m L乙醚-乙醇混合液进行超声脱脂1h与常规脱脂方法一致;热水提取1 h能除去糖类、淀粉的干扰;用2m L中性蛋白酶水解30 min能除去绝大部分蛋白质;方法回收率为98.31%～99.52%,日内RSD为1.21%～1.31%,日间RSD为1.39%。可以应用于烟草中总细胞壁物质含量的测定。In order to reduce the use of highly toxic reagent benzene and to eliminate interference of starch,protein and ash in the analysis of total cell wall substances content,this study used mixed solvent（ethyl ether：ethanol =1：15） treated sample by ultrasound degreasing.Thermostable α-amylase was added and sample was extracted with hot water.Protein was hydrolyzed by neutral protein enzyme,while residue was turned to ash at 510℃.Results showed that the 30 ml mixed solvent of ethanol and ethyl ether used for ultrasound degreasing was consistent with conventional degreasing method.Hot water extraction for 1h eliminated the interference of carbohydrate and starch,and hydrolysis with 2ml neutral protein enzyme for 30 min removed most protein in the sample.The recovery rate was 98.31%～99.52% with intraday RSD atl.21%～1.31% and interday RSD at 1.39%,indicating that the method is reliable and accurate and can be used for analysis of total cell wall substances content in leaf tobacco.

关 键 词：烟草 总细胞壁物质含量 除杂质 超声法 酶解法 

分 类 号：TS413[农业科学—烟草工业]