蚕丝中非蚕丝蛋白物质含量试验方法 Test method for non-protein content in silk  ICS  59.080.01 CCS  W 40   中华人民共和国国家标准 GB/T 44214—2024 2024-07-2 4发布 2025-02-0 1实施     国家市场监督管理总局 国家标准化管理委员会 发 布目　　次 前言   ·····································································································   Ⅲ 1     范围   ··································································································   1 2     规范性引用文件   ······················································································   1 3     术语和定义   ···························································································   1 4     原理   ··································································································   1 5     试剂和材料   ···························································································   1 6     仪器和设备   ···························································································   2 7     试验步骤   ······························································································   2      7.1     标准溶液配制   ····················································································   2      7.2     缓冲液配制   ······················································································   2      7.3     样品前处理   ······················································································   3      7.4     衍生化操作   ······················································································   3      7.5     萃取净化   ·························································································   3      7.6     测定   ······························································································   3 8     结果计算与表示   ······················································································   4      8.1     蚕丝中氨基酸含量分析   ··········································································   4      8.2     非蚕丝蛋白物质含量分析   ········································································   5 9     精密度   ································································································   5 10     试验报告   ····························································································   5 附录A （资料性）    四种氨基酸化合物信息   ····························································   6 GB/T 44214—2024     Ⅰ 前　　言 本文件按照GB/T 1.1—2020《标准化工作导则　第1部分：标准化文件的结构和起草规则》的规 定起草。 请注意本文件的某些内容可能涉及专利。本文件的发布机构不承担识别专利的责任。 本文件由中国纺织工业联合会提出。 本文件由全国丝绸标准化技术委员会（SAC/TC 401）归口。 本文件起草单位：浙江丝绸科技有限公司、黄山市信达丝线有限公司、浙江蚕缘家纺股份有限公 司、苏州太湖雪丝绸股份有限公司、嵊州陌桑高科股份有限公司、苏州大学、苏州市纤维检验院、南通 市纤维检验所、浙江巴贝领带有限公司、达利丝绸（浙江）有限公司、南充银海丝绸有限公司、四川安 泰茧丝绸集团有限公司、浙江省检验检疫科学技术研究院、杭州市质量技术监督检测院、上海市质量监 督检验技术研究院、浙江生态纺织品禁用染化料检测中心有限公司、江苏工程职业技术学院、中芯创惟 生物科技(杭州)有限公司、苏州市知识产权保护中心、绍兴市柯桥区中国轻纺城检验检测与标准化研 究院。 本文件主要起草人：吴国坚、伍冬平、陈思宇、孙晨晓、祁宁、刘雨、江朝辉、赵海浪、陈凤鸣、 刘伟、雷斌、倪学林、金丰、何国君、文朝钧、朱杨锐、杭志伟、李婷、曹石淼、陈志华、尹桂波、 潘璐璐、蒋伟、冯桂华、童珈珈。   GB/T 44214—2024     Ⅲ 蚕丝中非蚕丝蛋白物质含量试验方法 1　范围   本文件描述了测定蚕丝中非蚕丝蛋白物质含量的试验方法。 本文件适用于通过测定蚕丝中四种主要氨基酸（丝氨酸、甘氨酸、丙氨酸和酪氨酸）含量计算蚕丝 中非蚕丝蛋白物质的含量。 2　规范性引用文件   下列文件中的内容通过文中的规范性引用而构成本文件必不可少的条款。其中，注日期的引用文 件，仅该日期对应的版本适用于本文件；不注日期的引用文件，其最新版本（包括所有的修改单）适用 于本文件。 GB/T 6682　分析实验室用水规格和试验方法 GB/T 8170　数值修约规则与极限数值的表示和判定 GB/T 9995　纺织材料含水率和回潮率的测定　烘箱干燥法 3　术语和定义   下列术语和定义适用于本文件。 3.1　 非蚕丝蛋白物质　non-protein content 蚕丝经水解后，未分解成氨基酸的物质总量，包括超过正常范围的游离NH4+的量。 4　原理   蚕丝主要由丝素和丝胶组成，其在高温酸性溶液中可水解成游离氨基酸，与异硫氰酸苯酯反应后， 生成的氨基酸衍生物在紫外波长处有强吸收峰，经色谱柱分离后，采用高效液相色谱仪，对蚕丝中四种 主要氨基酸（丝氨酸、甘氨酸、丙氨酸、酪氨酸）进行测定。因蚕丝中氨基酸含量较为稳定，通过测定 蚕丝蛋白水解后氨基酸的含量，经过计算可得出蚕丝中非蚕丝蛋白物质的含量。 注：目前蚕丝中非蛋白物质主要有以甲基丙烯酰胺为单体的接枝聚合物。 5　试剂和材料   5.1　浓盐酸：优级纯。 5.2　乙腈：色谱纯。 5.3　异硫氰酸苯酯：纯度≥98%。 5.4　三乙胺：分析纯。 5.5　正己烷：色谱纯。 5.6　酸解溶液：6 mol/L 盐酸（5.1）溶液。 5.7　异硫氰酸苯酯﹘乙腈溶液（1 mol/L）：移取1.3 mL异硫氰酸苯酯于10 mL容量瓶中，用乙腈定容 GB/T 44214—2024     1 至刻度，混匀。 5.8　三乙胺﹘乙腈溶液（1 mol/L）：移取1.4 mL三乙胺于10 mL容量瓶中，用乙腈定容至刻度， 混匀。 5.9　水：符合GB/T 6682 二级水及以上要求。 5.10　氮气：纯度为99.99%。 5.11　氨基酸标准物质：丝氨酸（Ser）、甘氨酸（Gly）、丙氨酸（Ala）和酪氨酸（Tyr），化合物信 息见附录A，纯度均≥98%。 5.12　磷酸：纯度≥85%。 5.13　0.45 μm水相滤膜。 5.14　0.45 μm有机相滤膜。 6　仪器和设备   6.1　分