title: Validation of a confirmatory method for lipophilic marine toxins in shellfish using UHPLC-HR-Orbitrap MS.
abstract: Lipophilic marine toxins are produced by harmful microalgae and can accumulate in edible filter feeders such as shellfish, leading to an introduction of toxins into the human food chain, causing different poisoning effects. During the last years, analytical methods, based on liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS), have been consolidated by interlaboratory validations. However, the main drawback of LC-MS/MS methods remains the limited number of compounds that can be analyzed in a single run. Due to the targeted nature of these methods, only known toxins, previously considered during method optimization, will be detected. Therefore in this study, a method based on ultra-high-performance liquid chromatography coupled to high-resolution Orbitrap mass spectrometry (UHPLC-HR-Orbitrap MS) was developed. Its quantitative performance was evaluated for confirmatory analysis of regulated lipophilic marine toxins in shellfish flesh according to Commission Decision 2002/657/EC. Okadaic acid (OA), dinophysistoxin-1 (DTX-1), pectenotoxin-2 (PTX-2), azaspiracid-1 (AZA-1), yessotoxin (YTX), and 13-desmethyl spirolide C (SPX-1) were quantified using matrix-matched calibration curves (MMS). For all compounds, the reproducibility ranged from 2.9 to 4.9 %, repeatability from 2.9 to 4.9 %, and recoveries from 82.9 to 113 % at the three different spiked levels. In addition, confirmatory identification of the compounds was effectively performed by the presence of a second diagnostic ion ((13)C). In conclusion, UHPLC-HR-Orbitrap MS permitted more accurate and faster detection of the target toxins than previously described LC-MS/MS methods. Furthermore, HRMS allows to retrospectively screen for many analogues and metabolites using its full-scan capabilities but also untargeted screening through the use of metabolomics software.
authors.affiliation: Faculty of Veterinary Medicine, Department of Veterinary Public Health and Food Safety, Laboratory of Chemical Analysis, Ghent University, Salisburylaan 133, 9820, Merelbeke, Belgium.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Orellana
authors.forename: Julie
authors.initials: J
authors.lastname: Vanden Bussche
authors.forename: Lieven
authors.initials: L
authors.lastname: Van Meulebroek
authors.forename: Michiel
authors.initials: M
authors.lastname: Vandegehuchte
authors.forename: Colin
authors.initials: C
authors.lastname: Janssen
authors.forename: Lynn
authors.initials: L
authors.lastname: Vanhaecke
mesh_terms: D000818:Animals; D049872:Bivalvia; D002138:Calibration; D049873:Cardiidae; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D049896:Crassostrea; D005506:Food Contamination / Q000032:analysis*; D057230:Limit of Detection; D008387:Marine Toxins / Q000032:analysis*; D008403:Mass Screening; D013058:Mass Spectrometry / Q000379:methods*; D049878:Mytilus; D019319:Okadaic Acid / Q000032:analysis; D010839:Phytoplankton; D011786:Quality Control; D015203:Reproducibility of Results; D012758:Shellfish
chemical_list: D008387:Marine Toxins; D019319:Okadaic Acid