新型脂肪胺荧光标记试剂的合成及小分子 脂肪胺的衍生检测
目的 合成一种新的以荧光素为荧光团, N-羟基琥珀酰亚胺活性酯为反应基团的脂肪胺荧光标记试剂, 并建立以新试剂标记短链脂肪胺并用高效液相色谱-荧光检测法进行分离的方法。方法 在pH 8.5的Na2B4O7-H3BO3缓冲溶液中, 25 ℃下, 衍生试剂浓度为50 μmol/L, 衍生反应进行10 min, 衍生物峰面积即可达到最大且稳定。结果 在优化的条件下, 建立的检测方法的线性范围为0.01~2 μmol/L, 线性回归系数＞0.999, 检测限达到0.1~1 μg/L。结论 衍生后的产物利用高效液相色谱-荧光检测法, 可以使得6种小分子脂肪胺得到有效分离, 方法具有衍生速度快, 灵敏度高等优点。
Objective To synthesize a new aliphatic amine labeling reagent, using fluorescein as fluorophore and N-hydroxysuccinimidyl acetive ester as reaction group, and establish a separation method by labeling short chain aliphatic amine with new reagent and using high performance liquid chromatography- fluorescence detection. Methods The derivatization reaction was achieved with the buffer of Na2B4O7-H3BO3 at pH 8.5, the reaction temperature at 25 ℃, the reagent solution at 50 μmol/L and the reaction time of 10 min. Results The linear range from 0.01 μmol/L to 2 μmol/L was observed for the proposed method. The limit of detection (LOD) of developed method was between 0.1 μg/L and 1 μg/L with the linear regression coefficient above 0.999. Conclusion The baseline separation of 6 small molecule aliphatic amine derivatives is achieved using optimized HPLC fluorescence detection. The method has the advantages of high derivative speed and sensitivity.
标题：新型脂肪胺荧光标记试剂的合成及小分子 脂肪胺的衍生检测
英文标题：Synthesization of novel aliphatic amine fluorescence labeling reagents and derivative determination of micromolecule aliphatic amine
作者：
肖玲艳 通标标准技术服务有限公司宁波分公司
李赛花 通标标准技术服务有限公司宁波分公司
马明 宁波出入境检验检疫局技术中心;宁波检验检疫科学技术研究院
肖道清 宁波出入境检验检疫局技术中心;宁波检验检疫科学技术研究院
陈丹超 宁波出入境检验检疫局技术中心
俞雄飞 宁波出入境检验检疫局技术中心
中文关键词:脂肪胺,荧光标记,高效液相色谱,
英文关键词:aliphatic amine,fluorescence labeling,high performance liquid chromatography,
发表日期：2015-11-18
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