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
We developed a rapid and highly sensitive analytical method for chlorophylls and carotenoids derived from marine phytoplankton using ultra-high performance liquid chromatography (UHPLC). High-performance liquid chromatography (HPLC) has been widely used in phytoplankton pigment analysis since the 19...
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Online Access: | http://hdl.handle.net/2115/67627 https://doi.org/10.1016/j.marchem.2015.07.010 |
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fthokunivhus:oai:eprints.lib.hokudai.ac.jp:2115/67627 2023-05-15T15:44:00+02:00 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 Suzuki, Koji Kamimura, Akiko Hooker, Stanford B http://hdl.handle.net/2115/67627 https://doi.org/10.1016/j.marchem.2015.07.010 eng eng http://hdl.handle.net/2115/67627 Marine Chemistry, 176: 96-109 http://dx.doi.org/10.1016/j.marchem.2015.07.010 © 2015, Elsevier. Licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ http://creativecommons.org/licenses/by-nc-nd/4.0/ CC-BY-NC-ND Keywords: Algal pigments Chlorophylls Carotenoids Marine phytoplankton Ultra-high performance liquid chromatography (UHPLC) First derivative spectrum chromatogram (FDSC) 465 article (author version) fthokunivhus https://doi.org/10.1016/j.marchem.2015.07.010 2022-11-18T01:03:36Z We developed a rapid and highly sensitive analytical method for chlorophylls and carotenoids derived from marine phytoplankton using ultra-high performance liquid chromatography (UHPLC). High-performance liquid chromatography (HPLC) has been widely used in phytoplankton pigment analysis since the 1980’s for estimating the abundance, composition, and photosynthetic physiology of natural algal assemblages or laboratory cultures. However, the run-time of the HPLC analyses is generally ca. 30 min or more, which is time-consuming for analysts. Our UHPLC technique enabled us to complete the separations of chlorophylls and carotenoids from marine phytoplankton within 7 min with similar resolution as conventional HPLC methods. The analytical method was tested on authentic pigment standards, marine phytoplankton cultures, and field samples that were collected from the central tropical and subarctic Pacific plus the neritic Bering Sea. Critical pigment pairs that generally co-eluted as a single peak were successively resolved by obtaining the first derivative spectrum chromatograms (FDSCs) with a photodiode array (PDA) detector based on differences in pigment absorption spectra, e.g., chlorophyll (Chl) c2 and Mg 2,4 divinyl (DV) pheoporphyrin a5 monomethyl ester (MgDVP), as well as DVChl b and Chl b. Because the maximum injection volume of UHPLC is generally lower than that of HPLC to minimize the unwanted broadening of chromatographic peaks, the detection sensitivity needed to be increased, especially for oligotrophic seawater samples with low pigment concentration. To overcome this sensitivity issue, a PDA detector equipped with an 85 mm path length capillary cell was used with a fluorescence detector. As a result, the limit of quantitation (LOQ) as determined by absorbance was of the order of 0.1 ng for chlorophylls and carotenoids. Furthermore, a bead-beating technique using N,N-dimethylformamide (DMF) and zirconia beads was used to minimize the volume of the organic solvent utilized for pigment extraction. Our UHPLC ... Article in Journal/Newspaper Bering Sea Subarctic Hokkaido University Collection of Scholarly and Academic Papers (HUSCAP) Bering Sea Pacific Marine Chemistry 176 96 109 |
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Open Polar |
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Hokkaido University Collection of Scholarly and Academic Papers (HUSCAP) |
op_collection_id |
fthokunivhus |
language |
English |
topic |
Keywords: Algal pigments Chlorophylls Carotenoids Marine phytoplankton Ultra-high performance liquid chromatography (UHPLC) First derivative spectrum chromatogram (FDSC) 465 |
spellingShingle |
Keywords: Algal pigments Chlorophylls Carotenoids Marine phytoplankton Ultra-high performance liquid chromatography (UHPLC) First derivative spectrum chromatogram (FDSC) 465 Suzuki, Koji Kamimura, Akiko Hooker, Stanford B 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 |
topic_facet |
Keywords: Algal pigments Chlorophylls Carotenoids Marine phytoplankton Ultra-high performance liquid chromatography (UHPLC) First derivative spectrum chromatogram (FDSC) 465 |
description |
We developed a rapid and highly sensitive analytical method for chlorophylls and carotenoids derived from marine phytoplankton using ultra-high performance liquid chromatography (UHPLC). High-performance liquid chromatography (HPLC) has been widely used in phytoplankton pigment analysis since the 1980’s for estimating the abundance, composition, and photosynthetic physiology of natural algal assemblages or laboratory cultures. However, the run-time of the HPLC analyses is generally ca. 30 min or more, which is time-consuming for analysts. Our UHPLC technique enabled us to complete the separations of chlorophylls and carotenoids from marine phytoplankton within 7 min with similar resolution as conventional HPLC methods. The analytical method was tested on authentic pigment standards, marine phytoplankton cultures, and field samples that were collected from the central tropical and subarctic Pacific plus the neritic Bering Sea. Critical pigment pairs that generally co-eluted as a single peak were successively resolved by obtaining the first derivative spectrum chromatograms (FDSCs) with a photodiode array (PDA) detector based on differences in pigment absorption spectra, e.g., chlorophyll (Chl) c2 and Mg 2,4 divinyl (DV) pheoporphyrin a5 monomethyl ester (MgDVP), as well as DVChl b and Chl b. Because the maximum injection volume of UHPLC is generally lower than that of HPLC to minimize the unwanted broadening of chromatographic peaks, the detection sensitivity needed to be increased, especially for oligotrophic seawater samples with low pigment concentration. To overcome this sensitivity issue, a PDA detector equipped with an 85 mm path length capillary cell was used with a fluorescence detector. As a result, the limit of quantitation (LOQ) as determined by absorbance was of the order of 0.1 ng for chlorophylls and carotenoids. Furthermore, a bead-beating technique using N,N-dimethylformamide (DMF) and zirconia beads was used to minimize the volume of the organic solvent utilized for pigment extraction. Our UHPLC ... |
format |
Article in Journal/Newspaper |
author |
Suzuki, Koji Kamimura, Akiko Hooker, Stanford B |
author_facet |
Suzuki, Koji Kamimura, Akiko Hooker, Stanford B |
author_sort |
Suzuki, Koji |
title |
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 |
title_short |
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 |
title_full |
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 |
title_fullStr |
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 |
title_full_unstemmed |
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 |
title_sort |
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 |
url |
http://hdl.handle.net/2115/67627 https://doi.org/10.1016/j.marchem.2015.07.010 |
geographic |
Bering Sea Pacific |
geographic_facet |
Bering Sea Pacific |
genre |
Bering Sea Subarctic |
genre_facet |
Bering Sea Subarctic |
op_relation |
http://hdl.handle.net/2115/67627 Marine Chemistry, 176: 96-109 http://dx.doi.org/10.1016/j.marchem.2015.07.010 |
op_rights |
© 2015, Elsevier. Licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ http://creativecommons.org/licenses/by-nc-nd/4.0/ |
op_rightsnorm |
CC-BY-NC-ND |
op_doi |
https://doi.org/10.1016/j.marchem.2015.07.010 |
container_title |
Marine Chemistry |
container_volume |
176 |
container_start_page |
96 |
op_container_end_page |
109 |
_version_ |
1766378223446261760 |