Simultaneous determination and screening of five pigments in marine phytoplanktons by high performance liquid chromatography-triple quadrupole mass spectrometry

Abstract

A quantitative method based on high performance liquid chromatography coupled with electrospray ionization tandem triple-quadrupole mass spectrometry (HPLC-ESI-QqQ-MS) has been established for five pigments in marine phytoplanktons. The HPLC method used ternary solvent systems and a reversed-phase C16-amide column. In addition, methanol, acetonitrile and aqueous ammonium acetate were used as mobile phases. Five pigments (chlorophyll a, chlorophyll b, β, β-carotene, lutein and fucoxanthin) were quantified in selective reaction mode. As results, good linear relationships were achieved between the concentrations and the peak areas of the five pigment standards. And their correlation coefficients (r2) were higher than 0.996. The recoveries of the pigment standards were between 82.77% and 99.83%. The inter-day and intra-day precisions were lower than 5% (n = 5). The detection limits of the pigments for this method were between 0.02 and 0.16 μg/L and the quantification limits were in the range from 0.06 to 0.54 μg/L. According to the above method, eleven algae (Heterosigma akashiwo (NMBRah03-2), Heterosigma akashiwo (NMBRah03-2-2), Karlodinium veneficum (NMBjah047-1), Prorocentrum minimum ( NMBjah042), Nannochloropsis oceanic (NMBluh014), Chlorella pyrenoidosa (NMBluh015-1), Pleurochrysis sp. (NMBjih026-1), Prymnesium sp. (NMBjih029), Skeletonema costatum (NMBguh004-1), Thalassiosira weiss- flogii (NMBguh021) and Thalassiosira pseudonana) (NMBguh005)) have been investigated for comparing the pigment distributions. The method is sensitive, accurate, reproducible, and useful for the study of alga compositions.