Environmental Protection Agency, Office of Research and Development, National Health and Environmental Effects Research Laboratory, Atlantic Ecology Division, 27 Tarzwell Dr., Narragansett, RI 02882, USA.
J Chromatogr A. 2011 Jun 3;1218(22):3432-8. doi: 10.1016/j.chroma.2011.03.058. Epub 2011 Apr 1.
In this study, a reverse-phase HPLC method incorporating a ternary solvent system was developed to analyze most polar and non-polar chlorophylls and carotenoids present in phytoplankton. The method is based on an RP-C₁₆-Amide column and provided excellent peak resolution of most taxonomically important pigments and an elution profile different than C₈ or C₁₈ columns provide. Analysis of mixed pigment standards, extracts of phytoplankton monocultures, and field samples showed that this method was able to resolve more than sixty pigments, ranging from very polar acidic chlorophylls to the non-polar hydrocarbon carotenes in less than 36 min. This included chlorophylls c₁, c₂ and c₃, divinyl chlorophylls a and b, the carotenoids lutein and zeaxanthin and some recently discovered pigments. The ability of this method to resolve divinyl chl b from monovinyl chl b and divinyl chl a from monovinyl chl a is particularly important for the quantification and identification of the marine cyanobacteria Prochlorococcus spp. in oceanic waters. The described protocol is sensitive and reproducible and can be used to assess the distribution and dynamics of major phytoplankton groups in marine and freshwater ecosystems.
在这项研究中，开发了一种反相高效液相色谱法，其中包含三元溶剂系统，用于分析浮游植物中存在的大多数极性和非极性叶绿素和类胡萝卜素。该方法基于 RP-C₁₆-Amide 柱，可提供大多数分类学上重要的色素的优异峰分辨率，并且洗脱轮廓与 C₈ 或 C₁₈ 柱不同。混合色素标准品、浮游植物单培养物提取物和现场样品的分析表明，该方法能够在不到 36 分钟的时间内分离出 60 多种色素，范围从非常极性的酸性叶绿素到非极性的类胡萝卜素。这包括叶绿素 c₁、c₂ 和 c₃、二乙烯基叶绿素 a 和 b、叶黄素和玉米黄质以及一些最近发现的色素。该方法能够将二乙烯基 chl b 与单乙烯基 chl b 分离，以及将二乙烯基 chl a 与单乙烯基 chl a 分离，这对于海洋水层中海洋蓝细菌 Prochlorococcus spp.的定量和鉴定特别重要。所描述的方案灵敏且可重现，可用于评估海洋和淡水生态系统中主要浮游植物群的分布和动态。
