BS EN 16204:2012 pdf download – Foodstuffs — Determination of lipophilic algal toxins (okadaic acid group toxins, yessotoxins, azaspiracids, pectenotoxins) in shellfish and shellfish products by LC-MS/MS This European Standard specifies a multi-reference method for the determination of lipophilic algal toxins (fatsoluble algal toxins produced by some dinoflagellates) in raw shellfish and shellfish products including cookedshellfish, by liquid chromatography coupled to tandem mass spectrometry LC-MSIMS [1],[2],[3].This methodhas been validated in an inter-laboratory study consisting of three parts via the analysis of both naturallycontaminated homogenates of blue mussel and spiked extracts of blue mussel, oyster and clam.For furtherinformation on the validation, see Annex A. Additional studies have investigated further matrices (see[4],[5]). The detection limit for toxins of the okadaic acid group, azaspiracids and pectenotoxins was determined to be6 ug/kg shelfish meat and for yessotoxins 10 ug/kg shellfish meat. Quantitative determination of okadaic acid (OA), pectenotoxin-2(PTX-2), azaspiracid-1(AZA-1) andyessotoxin (YTX) can be carried out directly by means of standard substances available commercially.Assuming an equal response factor, okadaic acid is used for the indirect quantitative determination of the twodinophysistoxins dinophysistoxin-1(DTX-1) and dinophysistoxin-2(DTX-2); likewise azaspiracid-1(AZA-1) isused for the indirect quantitative determination of azaspiracid-2(AZA-2) and azaspiracid-3(AZA-3), while YTXis used for homo-yessotoxin, 45-OH-yessotoxin and 45-OH-homo-yessotoxin, and PTX-2 for pectenotoxin-1(PTX-1). The limit of quantification (LOQ) for toxins of the okadaic acid group, azaspiracids and pectenotoxins wasdetermined to be 20 ug/kg shellfish meat and for yessotoxins 35 ug/kg shellfish meat. By means of hydrolysis [6], the esters of okadaic acid，DTX-1 and DTX-2 can also be determinedquantitatively as the corresponding free acids. 2Normative references The following documents，in whole or in part,are normatively referenced in this document and areindispensable for its application.For dated references, only the edition cited applies.For undated references,the latest edition of the referenced document (including any amendments) applies. EN ISO 3696:1995, Water for analytical laboratory use —Specification and test methods (ISo 3696:1987) 3Principle Remove the shellfish meat from the shell and homogenize the total shelfish meat. Extraction is carried outwith aqueous methanol (g = 80 % ).Separation is performed on a HPLC reverse-phase column provided with abinary gradient and detection is carried out by means of tandem mass spectrometry using triple quadrupoletechnology.The concentration of lipophilic toxins is determined by means of external calibration. 4Reagents lf not otherwise specified, reagents of analytical grade and solvents suitable for LC-MS/MS shall be used.Water shall be distilled in glass vessels or demineralised before use, or shall be of equivalent purity accordingto EN...
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