蛋白质的测定方法-中心实验室.docxVIP

蛋白质的测定方法Determination of protein修改记录Revision record序号No.修改时间Revision time修改页数Revision page描述Description版次Vision备注Remark123456789101．目的Purpose此份作业指导书为指导该实验提供依据，以确保实验数据的准确性。This operating instructions provide the basis for the guidance of the experiment to ensure the accuracy of the experimental data.2．适用范围Scope本方法适用于各类食品中蛋白质含量的测定。This method is applicable to the determination of the protein content of various types of food.3．相关文件Related Documents3.1 GB 5009.5-2010食品中蛋白质的测定Determination of protein in foods3.2 GB/T 5511 谷物和豆类 氮含量测定和粗蛋白质含量计算 凯氏法Cereals and legumes nitrogen content and calculation of crude protein content Kjeldahl method4．原理Principle食品中的蛋白质在催化加热条件下被分解，产生的氨与硫酸结合生成硫酸铵。碱化蒸馏使氨游离，用硼酸吸收后以硫酸或盐酸标准滴定溶液滴定，根据酸的消耗量乘以换算系数，即为蛋白质的含量。In food proteins are decomposed under heating conditions catalysis, binding of ammonia to form ammonium sulfate and sulfuric acid produced. Alkalized distilled ammonia free, after using boric acid absorption in sulfuric acid or hydrochloric acid standard volumetric titration, according to the consumption of acid multiplied by the conversion factor, that is, the protein content.5．试剂Reagents5.1 混合试剂：硫酸铜:硫酸钾=0.2:3；Mixed reagents: copper sulfate: potassium = 0.2:35.2 硫酸：98%；Sulfuric acid: 98%2%硼酸溶液：称取硼酸20g，加水溶解成1000ml；2% boric acid solution: Weigh boric acid 20g, dissolved in water to 1000ml.5.4 40%氢氧化钠：称取氢氧化钠200g，加水溶解成500ml；40% sodium hydroxide: NaOH weighed 200g, dissolved in water to 500ml.5.5混合指示液：1份0.1%甲基红乙醇溶液与5份0.1%溴甲酚绿乙醇溶液临用时混合；Mixed indicator solution: 1 part ethanol solution of 0.1% methyl red with 5 parts 0.1% bromocresol green mixed with ethanol temporary;5.6 C(1/2H2SO4)=0.1mol/L标准溶液或C(HCl)=0.1mol/L标准溶液。C (1/2H2SO4) = 0.1mol / L standard solution or a C (HCl) = 0.1mol / L standard solution.6．仪器Instrument6.1 凯氏定氮仪Kjeldahl apparatus6.2 分析天平Analytical Balance7．分析步骤Analysis step7.1 样品处理：称取充分均匀的固体试样0.20～2.00g、半固体试样2.00～5.00g或液体试样5.00～10.00ml于消化管中，加入混合试剂4.0g及10ml的硫酸，于消化炉消化,开始小心加热，待内容物全部炭化后，加大火力消化，直至液体蓝绿色出澄清透明后，继续加热0.5-1h。取下冷却后加入约40ml去离子水摇