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

许多种类的海洋无脊椎动物产生独特的亚氨基酸，即所谓的“鸦片”。冠瘿碱被认为是其厌氧糖酵解的代谢物，并且包括内消旋-阿拉诺平、D-斯特龙宾、D-牛磺平、β-阿拉诺平和D-章鱼碱。目前，有多种方法可用于定量测定观点。然而，没有一种方法能够同时确定这五种意见。这样的情况使得意见的研究有些麻烦。因此，我们尝试建立了一种同时测定五种阿片类药物的方法，通过邻苯二甲醛（OPA）衍生化和Sep-Pak Plus PS-1柱分离去除干扰的一级氨基酸。这种预处理使得有可能确定五个鸦片碱同时通过阳离子交换HPLC柱后NaClO/OPA检测。该方法的灵敏度在20 pmol - 1.0 nmol范围内。通过分析几种海洋动物中冠瘿碱的含量，证实了该方法对生物样品的适用性。

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.