Separation of chlorophylls and carotenoids from marine phytoplankton:: a new HPLC method using a reversed phase C8 column and pyridine-containing mobile phases
Separation of chlorophylls and carotenoids from marine phytoplankton:: a new HPLC method using a reversed phase C8 column and pyridine-containing mobile phases
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DOI：
10.3354/meps195029
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发表时间：
2000-01-01
影响因子：
2.5
通讯作者：
Garrido, JL
中科院分区：
文献类型：
--
作者：
Zapata, M;Rodríguez, F;Garrido, JL
建立了基于C-8反相柱和含吡啶流动相的高效液相色谱同时分离叶绿素和类胡萝卜素的方法。该方法具有足够的选择性，可以从chi a （MV类似物）中分离出单酰（MV）和双酰（DV）对极性叶绿素和DV叶绿素a (chl a)（原核生物原绿球藻的标记色素）。只有DV chi a/chl b对没有解析。这种对叶绿素的分辨能力以前只能通过使用聚合物C-18色谱柱与醋酸铵或含吡啶的流动相结合来实现。该方法还允许分离属于8个藻类类的分类群特异性类胡萝卜素，包括以前使用C-18柱的HPLC方法的一些关键色素对。该方法采用二元梯度，可用于低压和高压混合仪器。方法采用3套高效液相色谱系统进行可转移性检测。对于不同驻留体积的HPLC设备，只需稍微调整梯度剖面即可获得相似的结果。该方法对一些新发现的叶绿素和类胡萝卜素的选择性使其不仅适用于研究现场样品，而且适用于重新检查不同藻类类别的色素组成。
A high-performance liquid chromatographic (HPLC) method based on a reversed-phase C-8 column and pyridine-containing mobile phases was developed for the simultaneous separation of chlorophylls and carotenoids. The method is selective enough to resolve monovinyl (MV) and divinyl (DV) pairs of polar chlorophylls and DV chlorophyll a (chl a) (the marker pigment for the prokaryote Prochlorococcus marinus) from chi a (the MV analogue). Only the pair DV chi a/chl b was not resolved. This resolution capability for chlorophylls was only previously achieved using polymeric C-18 columns in combination with ammonium acetate or pyridine-containing mobile phases. The proposed method also allows the separation of taxon-specific carotenoids belonging to 8 algal classes, including some critical pigment pairs for previous HPLC methods using C-18 columns. The method employs a binary gradient, so it can be used with both low-pressure and high-pressure mixing instruments. Method transferability was tested using 3 HPLC systems. Only a slight adjustment of gradient profile was required to obtain similar results with HPLC equipment having different dwell volumes. The selectivity of the method towards some recently discovered chlorophyll and carotenoid pigments makes it especially suitable for studying not only field samples, but also for re-examining the pigment composition of different algal classes.