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METHOD ABSTRACTS105.2 / PARALYTIC SHELLFISH TOXINSANALYSIS OF DINOFLAGELATE DERIVED NEUROTOXINS IN BIVALVE MULLUSKS USINGHPLC POST-COLUMN FLUORESCENCE METHODThe paralytic shellsh toxins are a group of 18 secondary metabolites deposited in bivalve mollusks by dinoagelates. Dino-agelates blooms are seasonal, occurring during warm months. Since it is unpredictable whether an infestation will occur,the shellsh population should be regularly monitored for toxins. Ingestion of contaminated shellsh can lead to paralyticshellsh poisoning; a life-threatening illness.Mouse bioassay is the ofcial method of AOAC International, but the drawbacks associated with this method have led toexploration of chemical methods. The most common HPLC post-column method is to oxidize the separated toxins underalkaline conditions to a uorescent compound. Sullivan et al. 1 used this method to determine the gonyautoxins 1-6 (GTX1-6), saxitoxin (STX) and neosaxitoxin (neoSTX) but not the N-sulfocarbamoyl-11-hydroxysulfate toxins (C1-C4). Oshima etal. 2, 3 modied this method to determine the 3 toxin groups separately using isocratic elution with 3 different mobile phases.Further improvement by Jeffery van de Riet et al. 4, 5 has led to a shorter analysis time to determine the 3 groups of toxinsusing step gradient and a switching valve. This method abstract describes the use of Pickering Laboratories Pinnacle PCXpost-column derivatization system for the HPLC post-column determination of paralytic shell sh toxins.METHODEquipment:• LC with a Binary Pump• Fluorescence Detector• Pickering Laboratories dual reagentPinnacle PCX post-column derivatization unit(1153-1061 – 120 V, 1153-1062 – 240 V)• Agilent Technologies Zorbax Bonus RP column,3.5 µm, 4.6 x 150 mm (863668-901)Reagents:• Heptane Sulfonate• Phosphoric Acid• Ammonium Hydroxide• Acetonitrile• Periodic Acid• Sodium Hydroxide• Nitric AcidLC ConditionsSample Injection Volume: 10 µLFlow Rate: 0.8 mL/minMobile Phase:A. 11 mM heptane sulfonate, 5.5 mM phosphoric acid, adjustedto pH 7.1 with ammonium hydroxide.B. 11 mM heptane sulfonate, 16.5 mM phosphoric acid, 11.5 %acetonitrile, adjusted to pH 7.1 with ammonium hydroxide.Post-column ConditionsColumn Temperature: 40 °CReactor Volume: 1.0 mL (knitted)Reactor Temperature: 85 °CReagent Flow Rate: 0.4 mL/minReagents:1. 100 mM phosphoric acid, 5 mM periodic acid,adjusted to pH 7.8 with 5 M sodium hydroxide2. 0.75 M nitric acidDetection: Fluorometer ex: 330 nm, em: 390 nm
METHOD ABSTRACTSGTX4GTX1dcGTX3GTX5 dcGTX2neoSTXdcSTXSTXGTX2GTX30.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00Chromatogram of GTX and STX mixed working standardHPLC GRADIENT PROGRAMTIME (Min) %A %B0 100 07.9 100 08 0 10018.5 0 10018.6 100 024 100 0123REFERENCES:1) J.J.Sullivan et. alJ.Food Sci. 50(1) (1985) 26-29.2) Yasukatsu Oshima“Post-column Derivatization Liquid Chromatographic Method for Paralytic Shellfi sh Toxins.”Journal of AOAC International Vol.78, No.2, (1995) 528-532.3) Yasukatsu Oshima, et. alIn Mycotoxins and Phytotoxins “88, Elsevier Science Publishers,Amsterdam, The Netherlands, 319-326.4) Wade A. Rourke, et. al“Rapid Post-column Methodology for Determination of Paralytic Shellfi sh Toxins in Shellfi sh Tissue.”Journal of AOAC International Vol.91, No.3, (2008) 589-597.5) Jeffrey M. van de Riet, et. al“Liquid Chromatographic Post-column Oxidation Method for Analysis of Paralytic Shellfi sh Toxins inMussels, Clams, Scallops and Oysters: Single-Laboratory Validation:”Journal of AOAC International Vol.92, No. 6, (2009) 1690-1704.PRODUCT CATALOGPICKERING LABORATORIES10/11
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TABLE OF CONTENTSINTRODUCTION3 Cust
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INTRODUCTIONINSTALLATION & TRAINING
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“A LOT HAS CHANGED SINCE PRODUCIN
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INSTRUMENTS & HARDWAREFEATURE HIGHL
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INSTRUMENTS & HARDWARE• Indoor us
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INSTRUMENTS & HARDWARECRX400 POST-C
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CHROMATOGRAMS, COLUMNS & GUARDSPick
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CHROMATOGRAMS, COLUMNS & GUARDSREVE
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CHROMATOGRAMS, COLUMNS & GUARDS70-M
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CHROMATOGRAMS, COLUMNS & GUARDSHIGH
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CHROMATOGRAMS, COLUMNS & GUARDSSTAN
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CHROMATOGRAMS, COLUMNS & GUARDSSODI
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CHROMATOGRAMS, COLUMNS & GUARDSHIGH
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CHROMATOGRAMS, COLUMNS & GUARDSSTAN
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CHROMATOGRAMS, COLUMNS & GUARDSCARB
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CHROMATOGRAMS, COLUMNS & GUARDSSEPA
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CHROMATOGRAMS, COLUMNS & GUARDSALKI
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CHROMATOGRAMS, COLUMNS & GUARDSCOND
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CHROMATOGRAMS, COLUMNS & GUARDSAFLA
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CHROMATOGRAMS, COLUMNS & GUARDSVOGL
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CHROMATOGRAMS, COLUMNS & GUARDSANAL
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“PURE CHEMICALS DO WHAT THEY ARE
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REAGENTS & CHEMICALSSERAPREP & URIP
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REAGENTS & CHEMICALSRESTORECHROMATO
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REAGENTS & CHEMICALSCALIBRATION STA
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ELUANTSCHROMATOGRAPHIC GRADEPickeri
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APPLICATION KITSAmino Acid Analysis
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BECKMAN 6300/7300 COLUMNS & CHEMICA
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REPLACEMENT COMPONENTS, FITTINGS &
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Page 74 and 75: “THE LCTECH STORY”Pickering Lab
Page 76 and 77: LCTECH / GPC PRODUCT LINEGPC ULTRA
Page 78 and 79: LCTECH / GPC PRODUCT LINEUtilizing
Page 80 and 81: LCTECH / GPC PRODUCT LINE• Can be
Page 82 and 83: LCTECH / GPC PRODUCT LINEFEATURE HI
Page 84 and 85: LCTECH / GPC PRODUCT LINEFEATURE HI
Page 86 and 87: LCTECH / GPC PRODUCT LINE3Intensity
Page 88 and 89: LCTECH / GPC PRODUCT LINEEVA IIICON
Page 90 and 91: LCTECH / MYCOTOXIN PRODUCT LINEUV D
Page 92 and 93: LCTECH / MYCOTOXIN PRODUCT LINESAMP
Page 94 and 95: LCTECH / MYCOTOXIN PRODUCT LINEOTAR
Page 96 and 97: LCTECH / MYCOTOXIN PRODUCT LINEACCE
Page 98 and 99: “CUSTOMERS REALLY ONLY WANT THE C
Page 100 and 101: METHOD ABSTRACTS203.1 / MULTI-RESID
Page 102 and 103: METHOD ABSTRACTS208 / MULTI-RESIDUE
Page 104 and 105: METHOD ABSTRACTS328 / ANALYSIS OF A
Page 106 and 107: METHOD ABSTRACTS382 / 70-MIN AMINO
Page 108 and 109: METHOD ABSTRACTS205 / AMINOGLYCOSID
Page 110 and 111: METHOD ABSTRACTS102 / ANALYSIS OF B
Page 112 and 113: METHOD ABSTRACTS120 / BROMATE ANALY
Page 114 and 115: METHOD ABSTRACTS108 / POST-COLUMN L
Page 116 and 117: METHOD ABSTRACTS112 / EXPANDED HPLC
Page 118 and 119: METHOD ABSTRACTS325 / ANALYSIS OF F
Page 120 and 121: METHOD ABSTRACTS206 continued from
Page 126 and 127: METHOD ABSTRACTS104 / POLYETHER ANT
Page 128 and 129: METHOD ABSTRACTS124 / SULFONAMIDE A
Page 132 and 133: GENERAL TERMS & CONDITIONS1 CONTROL
Page 134: DISTRIBUTORSContinued from previous
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