蜂花粉总多酚的检测　福林酚试剂 比色法 Detection of total polyphenols in bee pollen—Colorimetric method using Folin-Ciocalteu reagent  ICS  65.140 CCS  B 47   中华人民共和国国家标准 GB/T 44349—2024 2024-09-2 9发布 2025-04-0 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     仪器设备   ······························································································   2 7     分析步骤   ······························································································   2      7.1     试样制备   ·························································································   2      7.2     试样前处理   ······················································································   2      7.3     空白试验   ·························································································   2      7.4     标准曲线的制作   ·················································································   2      7.5     试样测定   ·························································································   2 8     结果计算   ······························································································   3 9     定量限   ································································································   3 10     精密度   ·······························································································   3 附录A （资料性）    福林酚试剂（Folin﹘Ciocalteu reagent）配制方法   ································   4 参考文献   ··································································································   5 GB/T 44349—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。 本文件由中华全国供销合作总社提出。 本文件由全国蜂产品标准化技术委员会（SAC/TC 601）归口。 本文件起草单位：中国计量大学、安徽省王巢食品有限公司、广东正当年生物科技股份有限公司、 中科标准（宁德）科技有限公司、睿科集团（厦门）股份有限公司、深圳职业技术大学、秦皇岛海关技 术中心、阜阳职业技术学院、南京众谱生物科技有限公司、浙江省农业科学院、浙江省中医药研究院、 中国农业科学院蜜蜂研究所、青海省花宝蜂业股份合作公司、杭州天厨蜜源保健品有限公司、兰溪市产 品商品质量检测研究院、山东世通检测评价技术服务有限公司、陕西国蜂大健康产业有限公司、江西汪 氏蜜蜂园有限公司。 本文件主要起草人：李红亮、胡童霞、岳振峰、吴帆、崔宗岩、徐锦忠、曹联飞、韩亚超、 张敬群、王磊、赵光培、蒋先志、陈璇、李强强、董捷、李平、林影、李斌、车延年、陈云、汪玲。   GB/T 44349—2024     Ⅲ 蜂花粉总多酚的检测　福林酚试剂 比色法 1　范围   本文件描述了测定蜂花粉中总多酚的福林酚试剂比色方法。 本文件适用于蜂花粉中总多酚含量的测定。 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 5009.3—2016　食品安全国家标准　食品中水分的测定 GB/T 6682　分析实验室用水规格和试验方法。 3　术语和定义   下列术语和定义适用于本文件。 3.1　 蜂花粉　bee pollen 工蜂采集花粉，用唾液和花蜜混合后形成的物质。 [来源：GB/T 30359—2021，3.2] 4　原理   蜂花粉经75%乙醇溶液超声提取的总多酚物质的酚羟基在碱性条件下，与福林酚试剂中磷钨钼酸的 六价钨离子（W6+）反应生成蓝色的五价钨离子（W5+），可在765 nm波长处产生最大吸收。在一定浓 度范围内，其吸光度与总多酚类化合物含量成正比。与没食子酸标准曲线比较，可定量测定总多酚化合 物的含量。 5　试剂和材料   除非另有规定，仅使用分析纯试剂。 5.1　水，GB/T 6682，三级。 5.2　无水乙醇。 5.3　无水碳酸钠。 5.4　福林酚试剂（Folin﹘Ciocalteu reagent）：1 mol/L （或1 N），以棕色瓶分装，4 ℃避光保存。直 接购买或配制，配制方法见附录A。 5.5　75%（体积分数）乙醇溶液：加25 mL水至75 mL无水乙醇（5.2）中，混合摇匀。 5.6　碳酸钠溶液（10.0%）：称取10.00 g±0.02 g无水碳酸钠（5.3），加水溶解并稀释至100 mL， GB/T 44349—2024     1 摇匀。 5.7　没食子酸（C7H6O5）标准品：CAS号为149﹘91﹘7，纯度≥99.0%，或经国家认证并授予标准物质 证书的标准品。 5.8　没食子酸标准储备液（5.0 mg/mL）：准确称取0.500 0 g±0.000 1 g没食子酸标准品（5.7）于小烧 杯中，用约5 mL无水乙醇进行溶解，转移至100 mL容量瓶中，加水稀释并定容至刻度，摇匀。 6　仪器设备   6.1　分光光度计：波长范围为400 nm~800 nm，配1 cm标准比色皿。 6.2　离心机：配50 mL离心管容量转子，最高转速大于4 000 r/min。 6.3　电子天平：感量0.000 1 g和0.01 g各1台。 6.4　超声波清洗器：功率不小于200 W，水浴温度能恒定保持至50 ℃。 6.5　食品粉碎机：功率不小于200 W。 7　分析步骤  7.1　试样制备   取适量蜂花粉，用食品粉碎机完全研磨为粉状。 7.2　试样前处理   用电子天平称取研磨好的样品质量（m）为1.000 g±0.005 g的蜂花粉试样至50 mL具盖离心管中， 加入20 mL 75%乙醇溶液（5.