Spinochrome Identification and Quantification in Pacific Sea Urchin Shells, Coelomic Fluid and Eggs Using HPLC-DAD-MS.

Elena A Vasileva,Van T T Tran,Sergey A Fedoreyev,Natalia P Mishchenko,Hieu M N Vo

doi:10.3390/md19010021

Abstract

The high-performance liquid chromatography method coupled with diode array and mass spectrometric detector (HPLC-DAD-MS) method for quinonoid pigment identification and quantification in sea urchin samples was developed and validated. The composition and quantitative ratio of the quinonoid pigments of the shells of 16 species of sea urchins, collected in the temperate (Sea of Japan) and tropical (South-China Sea) climatic zones of the Pacific Ocean over several years, were studied. The compositions of the quinonoid pigments of sea urchins Maretia planulata, Scaphechinus griseus, Laganum decagonale and Phyllacanthus imperialis were studied for the first time. A study of the composition of the quinonoid pigments of the coelomic fluid of ten species of sea urchins was conducted. The composition of quinonoid pigments of Echinarachnius parma jelly-like egg membrane, of Scaphechinus mirabilis developing embryos and pluteus, was reported for the first time. In the case of Scaphechinus mirabilis, we have shown that the compositions of pigment granules of the shell epidermis, coelomic fluid, egg membrane, developing embryos and pluteus are different, which should enable a fuller understanding of the functions of pigments at different stages of life.

Highlights

The red granules discovered by MacMunn in the perivisceral fluid of the sea urchinEchinus esculentus over a century ago inspired research on the composition, structure, function and biological activity of the quinonoid pigments they contained, and still attract the attention of chemists and biologists [1]
In the HPLC-diode array detector (DAD)-MS method, we have developed HPLC conditions to successfully separate more than 20 quinonoid pigments of sea urchins during one short analysis
The developed HPLC-DAD-MS method and the obtained properties of spinochromes will be highly useful, for determination of composition and content of quinonoid pigments in various sea urchin samples, and for monitoring the quality of drugs and food additives based on sea urchin pigments, in addition to studying their stability

Summary

Introduction

The red granules discovered by MacMunn in the perivisceral fluid of the sea urchinEchinus esculentus over a century ago inspired research on the composition, structure, function and biological activity of the quinonoid pigments they contained, and still attract the attention of chemists and biologists [1]. The red granules discovered by MacMunn in the perivisceral fluid of the sea urchin. Pigment granules have been found in red spherule cells in the coelomic fluid (CF) for all species of sea urchins. Pigment granules were found in the jelly-like membrane covering the eggs of many, but not all, species of sea urchins. The number of granules and their location in the egg membrane (at the edge or in the centre) can be a species characteristic. Sea urchins of the order Clypeasteroida (Scaphechinus griseus, Scaphechinus mirabilis, and Echinarachnius parma) have from 15 to 40 pigment-containing chromatophores in the jelly-like egg membrane, the largest number being found in S. mirabilis eggs [5]. The brightly colored granules are detectable under an optical microscope in eggs, CF, gastrulating embryos and the larvae of S. mirabilis [6,7]

Methods

Results

Conclusion