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LC-MS/MS 检测卡尔毒素揭示了来自埃布罗三角洲（西北地中海）的海洋甲藻卡尔定鞭毛藻菌株中的新型变种。

LC-MS/MS Detection of Karlotoxins Reveals New Variants in Strains of the Marine Dinoflagellate Karlodinium veneficum from the Ebro Delta (NW Mediterranean).

机构信息

Alfred Wegener Institut-Helmholtz Zentrum für Polar- und Meeresforschung, Chemische Ökologie, Am Handelshafen 12, 27570 Bremerhaven, Germany.

Common Wadden Sea Secretariat, Virchowstr. 1, 26382 Wilhelmshaven, Germany.

出版信息

Mar Drugs. 2017 Dec 18;15(12):391. doi: 10.3390/md15120391.

DOI:10.3390/md15120391
PMID:29258236
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5742851/
Abstract

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 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 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 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.

摘要

建立了一种液相色谱-串联质谱（LC-MS/MS）方法，用于在选定反应监测（SRM）模式下检测和定量卡尔妥毒素。这种新方法基于对纯化的卡尔妥毒素（KcTx-1、KmTx-2、44-氧-KmTx-2、KmTx-5）、一种 Amphidinol（AM-18）以及未纯化的美国特拉华州海洋甲藻 品系 CCMP2936 大量培养物提取物的分析，该品系产生 KmTx-1 和 KmTx-3。SRM 方法对 KmTx-2 的检测限确定为 2.5 ng 柱上。记录了所有假定的卡尔妥毒素的碰撞诱导解离（CID）光谱，以呈现每种化合物的碎裂模式，用于其明确鉴定。从西班牙西北埃布罗三角洲的一个海湾分离的 K10 菌株的大量培养物产生了五种以前未报道的假定卡尔妥毒素，分子量为 1280、1298、1332、1356 和 1400 Da，与 KmTx-5 具有相似的片段。对埃布罗三角洲的 几个分离株的分析表明，卡尔妥毒素谱存在小规模的多样性，即 Fangar 湾的一个分离株产生 KmTx-5，而在附近但水文学上不同的 Alfacs 湾的几个分离株中发现了五种假定的新卡尔妥毒素。该 LC-MS/MS 方法的应用代表了在确定假定的卡尔妥毒素方面的渐进式进展，特别是在缺乏已知特定效力的完整纯化分析标准品谱的情况下。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/597f/5742851/4e9c0a4a61b9/marinedrugs-15-00391-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/597f/5742851/dea29c34e7ac/marinedrugs-15-00391-g001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/597f/5742851/b52cdc4369e9/marinedrugs-15-00391-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/597f/5742851/7a534ed15da9/marinedrugs-15-00391-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/597f/5742851/0e9e90fda323/marinedrugs-15-00391-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/597f/5742851/4e9c0a4a61b9/marinedrugs-15-00391-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/597f/5742851/dea29c34e7ac/marinedrugs-15-00391-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/597f/5742851/eb9346f79b00/marinedrugs-15-00391-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/597f/5742851/5d37954e3200/marinedrugs-15-00391-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/597f/5742851/0d77fe08a0d7/marinedrugs-15-00391-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/597f/5742851/b52cdc4369e9/marinedrugs-15-00391-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/597f/5742851/7a534ed15da9/marinedrugs-15-00391-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/597f/5742851/0e9e90fda323/marinedrugs-15-00391-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/597f/5742851/4e9c0a4a61b9/marinedrugs-15-00391-g008.jpg

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