LC-MS/MS Detection of Karlotoxins Reveals New Variants in Strains of the Marine Dinoflagellate Karlodinium veneficum from the Ebro Delta (NW Mediterranean)
A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the detection and quantitation of karlotoxins in the selected reaction monitoring (SRM) mode. This novel method was based upon the analysis of purified karlotoxins (KcTx-1, KmTx-2, 44-oxo-KmTx-2, KmTx-5), one amphid...
| Main Authors: | , , , , , , , , , , , |
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| Format: | info:eu-repo/semantics/article |
| Language: | English |
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MDPI
2020
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| Subjects: | |
| Online Access: | http://hdl.handle.net/10835/7325 |
| _version_ | 1789406477863092224 |
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| author | Krock, Bernd Busch, Julia A. Tillmann, Urban García Camacho, Francisco Sánchez Mirón, Asterio Gallardo Rodríguez, Juan José López Rosales, Lorenzo Andree, Karl B. Fernández Tejedor, Margarita Witt, Matthias Cembella, Allan D. Place, Allen R. |
| author_facet | Krock, Bernd Busch, Julia A. Tillmann, Urban García Camacho, Francisco Sánchez Mirón, Asterio Gallardo Rodríguez, Juan José López Rosales, Lorenzo Andree, Karl B. Fernández Tejedor, Margarita Witt, Matthias Cembella, Allan D. Place, Allen R. |
| author_sort | Krock, Bernd |
| collection | DSpace |
| description | A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the detection and quantitation of karlotoxins in the selected reaction monitoring (SRM) mode. This novel method was based upon the analysis of purified karlotoxins (KcTx-1, KmTx-2, 44-oxo-KmTx-2, KmTx-5), one amphidinol (AM-18), and unpurified extracts of bulk cultures of the marine dinoflagellate Karlodinium veneficum strain CCMP2936 from Delaware (Eastern USA), which produces KmTx-1 and KmTx-3. The limit of detection of the SRM method for KmTx-2 was determined as 2.5 ng on-column. Collision induced dissociation (CID) spectra of all putative karlotoxins were recorded to present fragmentation patterns of each compound for their unambiguous identification. Bulk cultures of K. veneficum strain K10 isolated from an embayment of the Ebro Delta, NW Mediterranean, yielded five previously unreported putative karlotoxins with molecular masses 1280, 1298, 1332, 1356, and 1400 Da, and similar fragments to KmTx-5. Analysis of several isolates of K. veneficum from the Ebro Delta revealed small-scale diversity in the karlotoxin spectrum in that one isolate from Fangar Bay produced KmTx-5, whereas the five putative novel karlotoxins were found among several isolates from nearby, but hydrographically distinct Alfacs Bay. Application of this LC-MS/MS method represents an incremental advance in the determination of putative karlotoxins, particularly in the absence of a complete spectrum of purified analytical standards of known specific potency. |
| format | info:eu-repo/semantics/article |
| id | oai:repositorio.ual.es:10835-7325 |
| institution | Universidad de Cuenca |
| language | English |
| publishDate | 2020 |
| publisher | MDPI |
| record_format | dspace |
| spelling | oai:repositorio.ual.es:10835-73252023-04-12T19:34:49Z LC-MS/MS Detection of Karlotoxins Reveals New Variants in Strains of the Marine Dinoflagellate Karlodinium veneficum from the Ebro Delta (NW Mediterranean) Krock, Bernd Busch, Julia A. Tillmann, Urban García Camacho, Francisco Sánchez Mirón, Asterio Gallardo Rodríguez, Juan José López Rosales, Lorenzo Andree, Karl B. Fernández Tejedor, Margarita Witt, Matthias Cembella, Allan D. Place, Allen R. phycotoxin ichthyotoxin HAB harmful algal bloom plankton Alfacs Bay Fangar Bay Mediterranean Sea A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the detection and quantitation of karlotoxins in the selected reaction monitoring (SRM) mode. This novel method was based upon the analysis of purified karlotoxins (KcTx-1, KmTx-2, 44-oxo-KmTx-2, KmTx-5), one amphidinol (AM-18), and unpurified extracts of bulk cultures of the marine dinoflagellate Karlodinium veneficum strain CCMP2936 from Delaware (Eastern USA), which produces KmTx-1 and KmTx-3. The limit of detection of the SRM method for KmTx-2 was determined as 2.5 ng on-column. Collision induced dissociation (CID) spectra of all putative karlotoxins were recorded to present fragmentation patterns of each compound for their unambiguous identification. Bulk cultures of K. veneficum strain K10 isolated from an embayment of the Ebro Delta, NW Mediterranean, yielded five previously unreported putative karlotoxins with molecular masses 1280, 1298, 1332, 1356, and 1400 Da, and similar fragments to KmTx-5. Analysis of several isolates of K. veneficum from the Ebro Delta revealed small-scale diversity in the karlotoxin spectrum in that one isolate from Fangar Bay produced KmTx-5, whereas the five putative novel karlotoxins were found among several isolates from nearby, but hydrographically distinct Alfacs Bay. Application of this LC-MS/MS method represents an incremental advance in the determination of putative karlotoxins, particularly in the absence of a complete spectrum of purified analytical standards of known specific potency. 2020-01-16T08:20:22Z 2020-01-16T08:20:22Z 2017-12-18 info:eu-repo/semantics/article 1660-3397 http://hdl.handle.net/10835/7325 en https://www.mdpi.com/1660-3397/15/12/391 Attribution-NonCommercial-NoDerivatives 4.0 Internacional http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess MDPI |
| spellingShingle | phycotoxin ichthyotoxin HAB harmful algal bloom plankton Alfacs Bay Fangar Bay Mediterranean Sea Krock, Bernd Busch, Julia A. Tillmann, Urban García Camacho, Francisco Sánchez Mirón, Asterio Gallardo Rodríguez, Juan José López Rosales, Lorenzo Andree, Karl B. Fernández Tejedor, Margarita Witt, Matthias Cembella, Allan D. Place, Allen R. LC-MS/MS Detection of Karlotoxins Reveals New Variants in Strains of the Marine Dinoflagellate Karlodinium veneficum from the Ebro Delta (NW Mediterranean) |
| title | LC-MS/MS Detection of Karlotoxins Reveals New Variants in Strains of the Marine Dinoflagellate Karlodinium veneficum from the Ebro Delta (NW Mediterranean) |
| title_full | LC-MS/MS Detection of Karlotoxins Reveals New Variants in Strains of the Marine Dinoflagellate Karlodinium veneficum from the Ebro Delta (NW Mediterranean) |
| title_fullStr | LC-MS/MS Detection of Karlotoxins Reveals New Variants in Strains of the Marine Dinoflagellate Karlodinium veneficum from the Ebro Delta (NW Mediterranean) |
| title_full_unstemmed | LC-MS/MS Detection of Karlotoxins Reveals New Variants in Strains of the Marine Dinoflagellate Karlodinium veneficum from the Ebro Delta (NW Mediterranean) |
| title_short | LC-MS/MS Detection of Karlotoxins Reveals New Variants in Strains of the Marine Dinoflagellate Karlodinium veneficum from the Ebro Delta (NW Mediterranean) |
| title_sort | lc-ms/ms detection of karlotoxins reveals new variants in strains of the marine dinoflagellate karlodinium veneficum from the ebro delta (nw mediterranean) |
| topic | phycotoxin ichthyotoxin HAB harmful algal bloom plankton Alfacs Bay Fangar Bay Mediterranean Sea |
| url | http://hdl.handle.net/10835/7325 |
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