Hantzsch反应衍生HPLC法测定酒中的微量甲醛.docVIP

稿件修改说明，稿件编号：ca130433稿件编号：ca130433antzsch反应衍生高效液相色谱法测定酒中的微量甲醛 徐莉莉，邰 超(，武俐，赵同谦, 韩 丹 （河南理工大学资源环境学院，焦作 454000） (摘 要 建立了Hantzsch反应衍生高效液相色谱法测定啤酒和白酒中微量甲醛的方法。试验考察了衍生条件对Hantzsch反应的影响，确定了最优衍生条件为pH 5.5，反应温度50℃，时间20 min，衍生剂乙酰丙酮（0.2%）0.5mL，浓度0.5mol L-1。该方法的线性关系良好，线性相关系数为0.99，相对标准偏差为4.7%，检出限为：0.2μg L-1。啤酒和白酒中的平均HCHO含量分别为106.3177.8μg L-1，加标回收率为81.2%～101.6%。 关键词 甲醛；高效液相色谱；Hantzsch反应；啤酒白酒文献Determination of Formaldehyde in Alcohol by High Performance Liquid Chromatography after Hantzsch Reaction Derivatization XU Li-li，TAI Chao，WUZHAO Tong-qian，HAN Dan (Institute of Resources and Environment, Henan Polytechnic University，Jiaozuo 454000Abstract: The high-performance liquid chromatography (HPLC) method was established to quantify formaldehyde in alcohol basing on Hantzsch reaction. The experimental conditions were optimized. Under the optimized condition, which were pH 5.5, reaction temperature 50 ℃, derivative agent acetyl acetone (0.2%) 0.5 mL, ammonium salt concentration 0.5 mol L-1 and reaction time 20 min, there was good linear relationship between the formaldehyde concentration and the peak area with the formaldehyde concentration ranging of 2 ~ 200 μg/L (R= 0.9989). The limit of detection of this method was 0.2 μg L-1 and the RSD was 4.7%. The recovery rate of formaldehyde was 81.2%～101.6%. method has been applied into the determination of formaldehyde in actual alcohol drink and the results were satisfactory. Keywords: formaldehyde; high performance liquid chromatograph; Hantzsch reaction; beer; liquor 为了延长保质期，降低成本,在啤酒生产的糖化过程中添加了甲醛作为稳定剂。而在啤酒的本身酵母发酵过程中，也会产生微量的甲醛。甲醛已被确定为可疑致癌物[1]，国家也明令啤酒行业禁止使用甲醛。目前，测定酒中微量甲醛的标准方法主要是乙酰丙酮比色法[2]和AHMT [3]，乙酰丙酮在甲醛含量低于0.2mg/L 时检出值准确性差，AHMT法在蒸馏过程中误差大，二者已很难满足甲醛的需要。另外已有不少研究报道了采用高效液相色谱法和气相色谱法检测食品、空气等样品中的甲醛[4-1]。乙酰丙酮只与甲醛和乙醛发生衍生反应，且在HPLC色谱条件下易分离出二者的衍生产物[12]。1 试验部分 1.1 仪器与试剂 Agilent 1200高效液相色谱、Milipore超纯水系统； 甲醛标准溶液1000 mg/L，临用时用超纯水稀释至所需不同浓度的标准工作溶液，冷藏于冰柜4℃；乙酰丙酮溶液，；溶液5 mol L-1；缓冲溶液NaH2PO4-Na2HPO4（0.2mol L，pH=6.8）；试剂均为分析纯，用水均为超纯水。 1.2 色谱分离条件 C18色谱柱；流动相：乙腈纯水（2