Kutlán D, Presits P, Molnár-Perl I
Institute of Inorganic and Analytical Chemistry, L. Eötvös University, Budapest, Hungary.
J Chromatogr A. 2002 Mar 8;949(1-2):235-48. doi: 10.1016/s0021-9673(01)01610-7.
A comprehensive evaluation of papers dealing with the HPLC quantitation of amines as o-phthaldialdehyde (OPA) derivatives has been given and discussed in details. The stability and characteristics of selected representatives of mono [methyl-, ethyl-, n-/isopropyl, n-/isobutyl-, tert.-butyl-, sec.-butyl-, isoamyl amines and ethanolamine), di- and polyamines (ethylenediamine, 1,2-propylenediamine, 1,3-propylenediamine, agmatine, tyramine, putrescine, cadaverine, histamine, spermine, spermidine, and bis(hexamethylene)triamine] have been investigated as their OPA/3-mercaptopropionic acid (MPA) and OPA/N-acetyl-L-cysteine (NAC) derivatives, from an analytical point of view, performing photodiode array and fluorescence detection, simultaneously. All amines having in their original structure the NH2-CH2-R moiety, in accord with the amino acids of the same structure, furnished more than one OPA derivative: their initially formed species transformed to further ones. On the basis of on-line HPLC-MS the transformed derivatives were proved to be the corresponding isoindoles that contain an additional OPA molecule. In order to achieve optimum analytical conditions derivatization reagents have been applied in different composition, in parallel. The OPA and the SH-group additive contents of the reagents have been varied in the mole ratios of OPA/MPA(NAC)=1:3 and OPA/MPA(NAC)=1:50. Data obtained proved that performing derivatizations by means of the OPA/MPA(NAC)=1:50 reagents resulted in two benefits: both the stability of derivatives could have been increased and the number of the transformed derivatives decreased. In case of aliphatic amines and in ethanolamine, the transformation of the initially formed derivative can be either quantitatively avoided as in the case of ethanolamine, or considerably decreased, below 1%, as in the cases of the other aliphatic monoamines investigated. As to the behavior of di- and polyamines the stability of derivatives has been considerably improved, the number of species have been decreased from four to two with the exception of spermidine. Stability values characterized both by the UV and fluorescence responses, as a function of the reaction time (from 90 s up to 6 h) have been given in details.
已对有关将胺类作为邻苯二甲醛（OPA）衍生物进行高效液相色谱定量分析的论文进行了全面评估，并进行了详细讨论。从分析角度出发，通过同时进行光电二极管阵列和荧光检测，研究了单胺[甲基胺、乙胺、正/异丙胺、正/异丁胺、叔丁胺、仲丁胺、异戊胺和乙醇胺]、二胺和多胺[乙二胺、1,2-丙二胺、1,3-丙二胺、胍丁胺、酪胺、腐胺、尸胺、组胺、精胺、亚精胺和双（六亚甲基）三胺]中选定代表物作为其OPA/3-巯基丙酸（MPA）和OPA/N-乙酰-L-半胱氨酸（NAC）衍生物的稳定性和特性。所有在其原始结构中具有NH2-CH2-R部分的胺类，与具有相同结构的氨基酸一致，会产生不止一种OPA衍生物：它们最初形成的物种会转化为其他物种。基于在线高效液相色谱-质谱分析，已证明转化后的衍生物是含有额外OPA分子的相应异吲哚。为了实现最佳分析条件，已并行应用了不同组成的衍生化试剂。试剂中OPA和SH基团添加剂的含量在OPA/MPA（NAC）=1:3和OPA/MPA（NAC）=1:50的摩尔比下变化。获得的数据证明，使用OPA/MPA（NAC）=1:50的试剂进行衍生化有两个好处：既可以提高衍生物的稳定性，又可以减少转化后衍生物的数量。对于脂肪族胺和乙醇胺，最初形成的衍生物的转化要么可以像乙醇胺那样定量避免，要么可以大幅降低至1%以下，就像所研究的其他脂肪族单胺的情况一样。至于二胺和多胺的行为，衍生物的稳定性已得到显著提高，除亚精胺外，物种数量已从四种减少到两种。已详细给出了以紫外和荧光响应表征的稳定性值作为反应时间（从90秒到6小时）的函数。
