一种利用纯培养藻株制备浮游植物光合色素标准品的方法研究

Preparation of Phytoplankton Pigments Standards from Pure Culture of Freshwater Algae

浮游植物色素组成分析可间接反映野外样品中浮游植物的相对丰度和组成。然而由于色素标准品的获得较为困难,制约了该方法的广泛应用。本研究开发了一种利用纯培养藻株制备浮游植物光合色素标准品的方法,包括Chlorophyll a、Chlorophyll b、Fucoxanthin、Lutein、Zeaxanthin、Alloxanthin、Neoxanthin、Violaxanthin、Peridinin。每种浮游植物均有其特征的的色素组成,提取纯培养藻株的色素,用液相色谱分离,通过比较各色素的出峰时间和吸收光谱,对色素进行定性分析。之后通过馏分收集获得各种色素的标准品,并用分光光度法确定各色素分离液的浓度。液相色谱分析结果显示各色素标准品纯度均在99%以上,各色素标准品的响应因子与购买的光合色素标准品有很好的相关性。

Analyzing the component and content of pigments in phytoplankton community enables the classification of strains into different phyla. However, the series of pigments standards are unavailable which restrains the application of this method. In this study, a method for the preparation of phytoplankton pigments standards from pure culture of freshwater algae was proposed, including chlorophyll a, chlorophyll b, fucoxanthin, lutein, zeaxanthin, alloxanthin, neoxanthin, violaxanthin and peridinin. Each phylum of phytoplankton has its typical composition of pigments. The pigments in each pure culture of phytoplankton were separated by high performance liquid chromatography （HPLC）. The chromatographic peaks were identified according to the typical composition of pigments in the phytoplankton, also the retention time and absorption spectra of each peak. After the peak of each pigment was qualitative identified, the corresponding fraction was collected. Pool collected fractions were used as standard solutions whose concentrations were determined by spectrophotometry. Purities of all the standard solutions were over 99% identified by HPLC. The response factors of the the standard solutions were well correlated with those in literatures.