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

Synthesization of novel aliphatic amine fluorescence labeling reagents and derivative determination of micromolecule aliphatic amine

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

中文摘要:

      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。

英文摘要:

      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.