用9-芴基甲基氯甲酸酯进行柱前衍生化并结合反相高效液相色谱法测定多胺
Determination of polyamines by precolumn derivatization with 9-fluorenylmethyl chloroformate and reverse-phase high-performance liquid chromatography.
作者信息
Sabri M I, Soiefer A I, Kisby G E, Spencer P S
机构信息
Center for Research on Occupational and Environmental Toxicology, Oregon Health Sciences University, Portland 97201.
出版信息
J Neurosci Methods. 1989 Jul;29(1):27-31. doi: 10.1016/0165-0270(89)90105-2.
A high-performance liquid chromatography (HPLC) method for the determination of picomole levels of polyamines (putrescine, spermidine, and spermine) is described. Amino groups in polyamines react with 9-fluorenylmethyl chloroformate (FMOC) to form stable and highly fluorescent derivatives which can be separated and quantitatively estimated by HPLC in about 12 min. The mean relative elution times (n = 14) for putrescine, spermidine and spermine are 4.21 +/- 0.02, 10.09 +/- 0.02 and 11.19 +/- 0.04 min, respectively. The method has been applied to determine polyamine concentration in rat dorsal root ganglia (DRG) without interference with endogenous amino acids. Polyamine content of individual rat DRG has been calculated and the values are as follows: putrescine, 36.8 +/- 2.01, spermidine, 1652 +/- 131.0 and spermine 388.5 +/- 38.4 pmol/DRG. Information on polyamine concentrations in DRG may be useful in understanding the mechanism of action of toxic chemicals on nervous system.
本文描述了一种用于测定皮摩尔水平多胺（腐胺、亚精胺和精胺）的高效液相色谱（HPLC）方法。多胺中的氨基与9-芴基甲基氯甲酸酯（FMOC）反应形成稳定且高荧光的衍生物，这些衍生物可通过HPLC在约12分钟内进行分离和定量测定。腐胺、亚精胺和精胺的平均相对洗脱时间（n = 14）分别为4.21±0.02、10.09±0.02和11.19±0.04分钟。该方法已应用于测定大鼠背根神经节（DRG）中的多胺浓度，且不受内源性氨基酸的干扰。已计算出单个大鼠DRG的多胺含量，结果如下：腐胺为36.8±2.01、亚精胺为1652±131.0、精胺为388.5±38.4 pmol/DRG。DRG中多胺浓度的信息可能有助于理解有毒化学物质对神经系统的作用机制。
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