Rapid and highly sensitive analysis of chlorophylls and carotenoids from marine phytoplankton using ultra-high performance liquid chromatography (UHPLC) with the first derivative spectrum chromatogram (FDSC) technique

摘要: Abstract We developed a rapid and highly sensitive analytical method for chlorophylls carotenoids derived from marine phytoplankton using ultra-high performance liquid chromatography (UHPLC). High-performance (HPLC) has been widely used in pigment analysis since the 1980s estimating abundance, composition, photosynthetic physiology of natural algal assemblages or laboratory cultures. However, run-time HPLC analyses is generally ca. 30 min more, which time-consuming analysts. Our UHPLC technique enabled us to complete separations within 7 min with similar resolution as conventional methods. The was tested on authentic standards, cultures, field samples that were collected central tropical subarctic Pacific plus neritic Bering Sea. Critical pairs co-eluted single peak successively resolved by obtaining first derivative spectrum chromatograms (FDSCs) photodiode array (PDA) detector based differences absorption spectra, e.g., chlorophyll (Chl) c 2 Mg 2,4 divinyl (DV) pheoporphyrin 5 monomethyl ester (MgDVP), well DVChl b Chl . Because maximum injection volume lower than minimize unwanted broadening chromatographic peaks, detection sensitivity needed be increased, especially oligotrophic seawater low concentration. To overcome this issue, PDA equipped an 85 mm path length capillary cell fluorescence detector. As result, limit quantitation (LOQ) determined absorbance order 0.1 ng carotenoids. Furthermore, bead-beating N , -dimethylformamide (DMF) zirconia beads utilized organic solvent extraction. can replace techniques, allows yield high-throughput data phytoplankton.