Development of new method for simultaneous quantitative determination of five opines found in marine invertebrates

许多海洋无脊椎动物群体会产生独特的亚氨基酸，也就是所谓的“opine”。鸦片类药物被认为是其厌氧糖酵解的代谢物，包括中-丙氨酸、D-司托宾、D-牛磺酸、β-丙氨酸和D-章鱼碱。目前，已有几种定量测定石油类药物的方法。然而，目前还没有一种方法能够同时确定这五种观点。这种情况使得对观点的研究变得有些麻烦。因此，我们尝试建立一种同时测定五种观点的方法，通过邻苯二甲醛(OPA)衍生化和随后的Sep-Pak Plus PS-1色谱柱分离去除干扰的主要氨基酸。这种前处理使阳离子交换高效液相色谱-柱后NaClO/OPA检测同时测定这五种观点成为可能。该方法的灵敏度在20pmol-1.0nmol范围内。通过对几种海洋动物体内OPINS含量的分析，证实了该方法在生物样品中的适用性。

Many groups of marine invertebrates produce unique imino acids so called 'opine'. Opines are considered as their metabolites of anaerobic glycolysis, and include meso- alanopine, D-strombine, D-tauropine, β-alanopine, and D-octopine. At present, several methods for quantitative determination of opines are available. However, there is no method capable of determining the five opines simultaneously. Such the situation makes the study of opine somewhat troublesome. Thus we attempted the development of a method determining the five opines simultaneously.Interfering primary amino acids were removed by o-phthalaldehyde (OPA) derivatization and the following fractionation using a Sep-Pak Plus PS-1 cartridge. This pre-treatment made possible to determine the five opines simultaneously by a cation-exchange HPLC with post-column NaClO/OPA detection. The sensitivity of this method is in the range 20 pmol - 1.0 nmol. The applicability of this method to biological samples was confirmed by analyzing the levels of opines in several marine animals.