title: Solid-phase extraction-based ultra-sensitive detection of four lipophilic marine biotoxins in bivalves by high-performance liquid chromatography-tandem mass spectrometry.
abstract: A solid-phase extraction (SPE) method for ultra-sensitive determination of four lipophilic marine biotoxins in bivalve samples by coupling high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS-MS) was developed. Azaspiracid-2 (AZA2), pectenotoxins-2, spirolide (SPX) and gymnodimine were simultaneously determined by HPLC-MS-MS in a positive multiple reaction monitoring mode. Separation was achieved on a reversed-phase C18 column with an acetonitrile-water gradient containing formic acid. During the analysis, solvent effects on the analytes were eliminated by using 1 : 1 water-methanol as dissolving solvent instead of pure methanol. Matrix effects in post-SPE extract and crude extract were seriously evaluated. Increased matrix effects in post-SPE extract countervailed the concentration purpose to some extent. The limits of detection of the SPE-HPLC-MS-MS method were determined to be in the range of 0.013-0.085 µg kg(-1), and the linear range of the method was in the range of 0.128-55.2 ng mL(-1) for the detected toxins. The proposed method was validated in terms of linearity (matrix-matched standard curves), precision, recovery, repeatability and limits of quantification. The recoveries of fortified samples at three different concentration levels were satisfactory, and the intra- and interday precisions were <7 and 10%, respectively.Several bivalve samples were analyzed to demonstrate the applicability of the proposed method. Different target toxins were detected in different kind of bivalves. Among them, AZA2 and SPX1 were first detected in Chinese shellfish. The levels of detected toxins were below the current European Union regulatory limits.
authors.affiliation: Ningbo Key Laboratory of Poison Research and Control, Ningbo Municipal Center for Disease Control and Prevention, Ningbo 315010, China flyxiangzi@yahoo.com flyxiangzi@gmail.com.
authors.forename: Lanyun
authors.initials: L
authors.lastname: Fang
authors.affiliation: Ningbo Key Laboratory of Poison Research and Control, Ningbo Municipal Center for Disease Control and Prevention, Ningbo 315010, China.
authors.forename: Xunping
authors.initials: X
authors.lastname: Yao
authors.affiliation: Ningbo Key Laboratory of Poison Research and Control, Ningbo Municipal Center for Disease Control and Prevention, Ningbo 315010, China.
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.affiliation: Ningbo Key Laboratory of Poison Research and Control, Ningbo Municipal Center for Disease Control and Prevention, Ningbo 315010, China.
authors.forename: Jige
authors.initials: J
authors.lastname: Li
mesh_terms: D000818:Animals; D049872:Bivalvia / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D057230:Limit of Detection; D016014:Linear Models; D008387:Marine Toxins / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification*; D015203:Reproducibility of Results; D012758:Shellfish; D052616:Solid Phase Extraction / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D008387:Marine Toxins