DB43/T2704—2023 43 湖 南 省 地 方 标 准ICS CCS 65.120 B 46 发 布 湖南省市场监督管理局饲料脱霉剂霉菌毒素吸附率测定 液相色谱串联质谱法 2023-08发布-22 2023-11实施-22Determination of mycotoxin adsorption rate of feed mycotoxin adsorbents by liquid chromatography-tandem mass spectrometry (LC-MS) method DB43/T 2704—2023 I 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 试剂与材料 ································ ································ ································ ············· 1 6 仪器设备 ································ ································ ································ ················ 2 7 试验步骤 ································ ································ ································ ················ 2 7.1 取样 ································ ································ ································ ··············· 2 7.2 混合吸附 ································ ································ ································ ········ 2 7.3 提取 ································ ································ ································ ··············· 2 7.4 霉菌毒素标准溶液净化液制备 ································ ································ ············· 3 7.5 测定 ································ ································ ································ ··············· 3 8 结果计算 ································ ································ ································ ················ 3 9 重复性 ································ ································ ································ ··················· 4 附录A（规范性） 霉菌毒素相关名称、分子式等信息对照表 ································ ················ 5 附录B（资料性） 霉菌毒素相关检测指标信息 ································ ································ ·· 6 DB43/T 2704—2023 II DB43/T 2704—2023 III 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。 本文件由湖南省农业农村厅提出。 本文件由湖南省农业标准化技术委员会归口。 本文件起草单位：湖南农业大学、江苏神力特生物科技有限公司、湖南生物机电职业技术学院、普 洱天澜农牧科技有限公司、湖南百宜科技集团有限公司。 本文件主要起草人：唐圣果、宋泽和、康萌、黄珂、黄正君、尹鑫、袁旭鹏、钱伟锋、罗文锋。 DB43/T 2704—2023 IV DB43/T 2704—2023 1 饲料脱霉剂霉菌毒素吸附率测定 液相色谱串联质谱法 1 范围 本文件规定了液相色 谱串联质谱法测定饲料脱霉剂霉菌毒素吸附率的原理、试剂与材料、仪器 设备、试验步骤、结果计算和重复性。 本文件适用于饲料脱霉剂对黄曲霉毒素（ AFB）B1、B2、G1和G2，玉米赤霉烯酮（ ZEA） ，伏马 毒素B1、B2、B3，脱氧雪腐镰刀菌烯醇（ DON） ，赭曲霉毒素 A、B和T-2毒素吸附率的测定。 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 14699.1 饲料采样 3 术语和定义 下列术语和定义适用于本文件。 3.1 霉菌毒素吸附率 mycotoxin adsorption rate 在pH=3的缓冲溶液中饲料脱霉剂对霉菌毒素的吸附比率，以吸附前后标准溶液中霉菌毒素浓度的 差值与吸附前霉菌毒素标准溶液浓度的比值表示。 4 原理 在pH=3模拟胃内环境的缓冲溶液中饲料脱霉剂与毒素标准液混合 1小时后，离心取上清液，测定 毒素的浓度，通过吸附前后标准溶液毒素浓度之差与吸附前标准溶液毒素浓度的比 值计算。 5 试剂与材料 所用试剂除另有规定外，均使用分析纯试剂： a） 水：GB/T 6682 ，一级； b） 浓盐酸； c） 氯化钾； d） 磷酸二氢钾； DB43/T 2704—2023 2 e） 十二水磷酸氢钠； f） 氯化钠； g） 甲酸：质谱纯； h） 乙腈：质谱纯； i） 甲醇：质谱纯； j） 霉菌毒素标准品：纯度及相关信息见附录 A； k） 盐酸溶液（ 2 mol/L） ：移取16.8 mL盐酸于1 L容量瓶中，加水定容至刻度，混匀后备用； l） 磷酸缓冲液（ pH=3） ：称取0.2 g氯化钾， 0.2 g磷酸二氢钾， 2.92 g十二水磷酸氢二钠， 8 g 氯化钠，加 950 mL水溶解，使用 2 mol/L盐酸溶液调整至 pH=3，最后用纯水定容到 1000 mL； m） 提取液：量取 159 mL水、840 mL乙腈和1 mL甲酸，混匀； n） 霉菌毒素储备液：按照附录 A称取黄曲霉毒素 B1、B2、G1、G2的标准品约 0.0003 g，称取赭 曲霉毒素 A、B，T-2毒素，玉米赤霉烯酮，脱氧雪腐镰刀菌烯醇，伏马霉素 B1、B2、B3的标 准品约0.003，置于10 mL容量瓶中，用乙腈溶解、定容，配制成混合霉菌毒素储备液；储备 液中黄曲霉毒素的浓度约为 30 μg/mL，其余霉菌毒素浓度约为 300 μg/mL； o） 霉菌毒素标准溶液：准确移取 100 μL霉菌毒素储备液于 10 mL