Cao Liwei, Wang Hong, Ma Ming, Zhang Huashan
Department of Chemistry, Wuhan University, Wuhan, PR China.
Electrophoresis. 2006 Feb;27(4):827-36. doi: 10.1002/elps.200500301.
An MEKC-LIF method using 6-oxy-(N-succinimidyl acetate)-9-(2'-methoxy-carbonyl) fluorescein (SAMF) newly synthesized in our lab as a labeling reagent for the separation and determination of eight typical biogenic amines was proposed. After careful study of the derivatization condition such as pH value, reagent concentration, temperature, and reaction time, derivatization reaction was accomplished as quickly as 10 min with stable yield. Optimal separation of SAMF-labeled amines was achieved with a running buffer (pH 9.3) containing 30 mM boric acid, 25 mM SDS, and 20% v/v ACN. The proposed method allowed biogenic amines to be determined with LODs as low as 0.25-2.5 nmol/L and RSD values from 0.4 to 4.5%. The present method has been successfully used to monitor biogenic amines in HeLa cells and fish samples. This study exploits the potential of MEKC-LIF with SAMF labeling as a tool for monitoring biogenic amines involved in complex physiological and behavioral processes in various matrices.
提出了一种使用在我们实验室新合成的6-氧代-（N-琥珀酰亚胺基乙酸酯）-9-（2'-甲氧基羰基）荧光素（SAMF）作为标记试剂的毛细管胶束电动色谱-激光诱导荧光（MEKC-LIF）方法，用于分离和测定8种典型生物胺。在仔细研究衍生化条件如pH值、试剂浓度、温度和反应时间后，衍生化反应可在10分钟内快速完成，产率稳定。使用含有30 mM硼酸、25 mM十二烷基硫酸钠（SDS）和20%（v/v）乙腈的运行缓冲液（pH 9.3）实现了SAMF标记胺的最佳分离。该方法能够测定生物胺，检测限低至0.25 - 2.5 nmol/L，相对标准偏差（RSD）值为0.4%至4.5%。本方法已成功用于监测HeLa细胞和鱼类样品中的生物胺。本研究利用了SAMF标记的MEKC-LIF作为监测各种基质中参与复杂生理和行为过程的生物胺的工具的潜力。
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