Dataset Information

Determination of neurotoxic shellfish poisoning toxins in shellfish by liquid chromatography-tandem mass spectrometry coupled with dispersive solid phase extraction.

ABSTRACT: An innovative method based on dispersive solid phase extraction (d-SPE) in conjunction with LC-MS/MS had been developed for the simultaneous quantitative determination of three brevetoxins (BTXs), which can result in neurotoxic shellfish poisoning (NSP), in shellfish. The toxins were extracted with a 50 % acetonitrile (v/v) and cleaned by alumina-neutral sorbent. After chromatographic separation on a C₁₈ column, the analytes were qualitatively and quantitatively detected using multiple reaction monitoring (MRM) in positive ionization mode. The created approach was validated by SANTE 11312/2021. The LOQs were 5 μg/kg for each toxin, below the advised regulatory limit of 800 μg BTX-2/kg. The mean recoveries of brevetoxins were in the range of 75.9 %-114.1 %, and the ranges of their intra- and inter-day precisions were 0.9-9.7 % and 0.6-7.2 %, respectively. The matrix effects for three BTXs in four shellfish matrices were in the range of 85.6 %-114.8 %. The method demonstrated great consistency and high sensitivity, and it can meet the requirements of daily monitoring.

SUBMITTER: Fang L

PROVIDER: S-EPMC10638005 | biostudies-literature | 2023 Nov

REPOSITORIES: biostudies-literature

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Determination of neurotoxic shellfish poisoning toxins in shellfish by liquid chromatography-tandem mass spectrometry coupled with dispersive solid phase extraction.

Fang Li L Qiu Fengmei F

Heliyon 20231027 11

An innovative method based on dispersive solid phase extraction (d-SPE) in conjunction with LC-MS/MS had been developed for the simultaneous quantitative determination of three brevetoxins (BTXs), which can result in neurotoxic shellfish poisoning (NSP), in shellfish. The toxins were extracted with a 50 % acetonitrile (v/v) and cleaned by alumina-neutral sorbent. After chromatographic separation on a C<sub>18</sub> column, the analytes were qualitatively and quantitatively detected using multipl ...[more]

PMID: 37954300

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Project description:Paralytic shellfish poisoning toxins (PSTs) are a family of more than 30 natural alkaloids synthesized by dinoflagellates and cyanobacteria whose toxicity in animals is mediated by voltage-gated Na(+) channel blocking. The export of PST analogues may be through SxtF and SxtM, two putative MATE (multidrug and toxic compound extrusion) family transporters encoded in PSTs biosynthetic gene cluster (sxt). sxtM is present in every sxt cluster analyzed; however, sxtF is only present in the Cylindrospermopsis-Raphidiopsis clade. These transporters are energetically coupled with an electrochemical gradient of proton (H(+)) or sodium (Na(+)) ions across membranes. Because the functional role of PSTs remains unknown and methods for genetic manipulation in PST-producing organisms have not yet been developed, protein structure analyses will allow us to understand their function. By analyzing the sxt cluster of eight PST-producing cyanobacteria, we found no correlation between the presence of sxtF or sxtM and a specific PSTs profile. Phylogenetic analyses of SxtF/M showed a high conservation of SxtF in the Cylindrospermopsis-Raphidiopsis clade, suggesting conserved substrate affinity. Two domains involved in Na(+) and drug recognition from NorM proteins (MATE family) of Vibrio parahaemolyticus and V. cholerae are present in SxtF/M. The Na(+) recognition domain was conserved in both SxtF/M, indicating that Na(+) can maintain the role as a cation anti-transporter. Consensus motifs for toxin binding differed between SxtF and SxtM implying differential substrate binding. Through protein modeling and docking analysis, we found that there is no marked affinity between the recognition domain and a specific PST analogue. This agrees with our previous results of PST export in R. brookii D9, where we observed that the response to Na(+) incubation was similar to different analogues. These results reassert the hypothesis regarding the involvement of Na(+) in toxin export, as well as the motifs L(398)XGLQD(403) (SxtM) and L(390)VGLRD(395) (SxtF) in toxin recognition.

Dataset Information

Determination of neurotoxic shellfish poisoning toxins in shellfish by liquid chromatography-tandem mass spectrometry coupled with dispersive solid phase extraction.

Publications

Determination of neurotoxic shellfish poisoning toxins in shellfish by liquid chromatography-tandem mass spectrometry coupled with dispersive solid phase extraction.

Similar Datasets

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