Jo Marie Cook Bureau of Chemical Residue Laboratories
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1 Jo Marie Cook Bureau of Chemical Residue Laboratories
2 Incident Command Do as your told! Wait and see not on our shores! Yet! Follow FDA lead Hesitation to spend $$$ FERN and ccap activation Bring up NOAA method Waiting for instruments, equipment, columns, standards Meetings, meetings, meetings Bring up Fluorescence Screen
3 Assure the Public! BP funding Safe Seafood Campaign 3 year surveillance PAH by LC Fluorescence 10% PAH by NOAA GC/MS DOSS by LC Triple 20% Metals by ICP/MS
4 Dollars Pounds Shrimp $25,836,726 16,432, Mullet $5,354,321 9,766,178 Grouper $17,718,249 6,721,722 Snapper $15,945,921 6,425,714 Mackerel, king $8,756,983 5,924,654 Blue crab $5,995, ,915, Spiny lobster $13,260,165 4,246,589 Stone crab claws $17, ,654,426 Oysters $6,134, ,524,821 TOTAL $116,655,465 59,611,878
5
6 Instrument & equipment intensive ASE, LC prep, Cool on-column. Time intensive 5 7 days for 20 samples Space intensive three hoods and prep area Resource intensive 3 chemists Hazardous reagents Standards unavailable GC/MSD lacks sensitivity and selectivity
7 Day 1 Receive Samples, log-in Prepare edible portion and freeze Day 2 Homogenize Frozen Automated Solvent Extraction (ASE) Day 3 Silica / Alumina Column Chromatography, Concentrate Size Exclusion HPLC Overnight Day 4 Concentrate Gas Chromatography Mass Spectroscopy (GC/MS) Day 5-6 Data evaluation Data reporting
8 Transforming a QuEChERS lab Weekly technical conference calls Learning about analytes & homologs Developing chromatography Eliminating contamination Measuring final 100uL volume Size exclusion clean-up very necessary Recovery of the SRM 1974b Using reporting template Nothing works with the power out!
9 Methylene Chloride hazards? Equipment in hood Snorkel instruments Hazardous waste Purple gloves!
10 Bake salts, rinse glassware Isolation from instruments pumps? Evaporation to 100uL by rotary evap, turbovap, KD, N2 blowdown
11 Day 1 Receive Samples, log-in Prepare edible portion and freeze Day 2 5 gm tissue + 5 ml H2O, Shake, 15mL ACN, Shake, MgSO4, Shake Centrifuge, filter and shoot Day 3 Data evaluation Data reporting Day 4 Day 5-6
12 FL detector do we have any of those? Faster, easier but no confirmation, no homologs 14 ppb of naphthalene interference Column dies peaks come and go 30% increase in signal for benzo[a]pyrene over 12 hours
13 ACN Syringe Filter Plastic Syringe Beads Wash Fisher (polypropylene) Blue Agilent Red Agilent Blue Bead Syringe Salts
14 Day 1 Receive Samples, log-in Prepare edible portion and freeze Day 2 5 gm tissue + 5 ml H2O, d34-doss Int. Std, Shake, 15mL ACN, Shake, MgSO4 & NaAc, Shake Centrifuge, filter, dilute 1/5 in H2O and shoot Day 3 Data evaluation Data reporting Day 4 Day 5-6
15 FCC NOAA Florida Instrument Applied Waters Quattro Waters Quattro Biosystems 5500 Micro Premier Gradient 45 95% CAN % CAN Isocratic 50% CAN 0.5 ml/min 0.3 ml/min 0.3mL/min Column ACE 3 C-18 18, Zorbax RRHD SB C-18 Acquity UPLC C-18 50mm X 3.0mm x 3um 50mm x 2.1mm x 1.8 um 50mm x 1.0mm x 1.7 um Sample Dilution 1/5 in ACN 1/5 in ACN 1/5 in H2O LOD 1 ng/g 15 ng/g 15 ng/g
16 FCC 50 ng/ml min NOAA Florida 50 ng/ml 1 ng/ml min min 1 ng/ml 5 ng/ml 1 ng/ml
17 Day 1 Receive Samples, log-in Prepare edible portion and freeze Day 2 10 gm tissue + 5 ml H2O, C13-PAH Int. Std, Shake, 10mL Ethyl Acetate, Shake, MgSO4 & NaCL, Shake Exchange to hexane, Silica gel clean-up with 3:1 hexane/methylene chloride Solvent exchange to iso-octane, GC/MS Day 3 Data evaluation Data reporting
18 UCT Tubes Fisher Polypropylene Low Standard
19 GC/MSD Cool On-column DB-5 60 x 0.25 x 0.25 GC/Triple PTV Restek RXI-175Sil-ms
20 GC/MSD Cool On-column DB-5 60 x 0.25 x 0.25 GC/Triple PTV Restek RXI-175Sil-ms b k j
21 Collection Number of Samples Dates: Number Number Number Range of 8/1/2010 without with exceeding Findings 5/1/2011 Total Detections Detections LOC* (mg/kg) Finfish ND Shrimp ND Trace** Oysters ND Trace Crabs ND Lobster ND Clams ND
22 DON T UNDER ESTIMATE CONSUMER CONCERNS FIT FOR PURPOSE Screening methods, if you must Quantitative MS methods, if efficient Research methods for scientific verification BE PREPARED Retain common instruments, capabilities Adopt flexible methods
23 Jo Marie Cook
24 Large samples are homogenized in Robo-Coupes Smaller samples are homogenized with probe blender Dry samples Add sodium sulfate & mix immediately Add magnesium sulfate Add to ASE cells Transfer to cells Add glass-fiber filters Run ASE overnight Load cells above Collect extracts in bottles below
25 Silica / Alumina Column Clean-up Removes polar compounds Concentrate Turbovap or column heaters or Kaderna-Danish Size Exclusion Chromatography Overnight Removes heavy weight compounds like lipids
26 Concentrate to 100uL Start GC/MS analysis 60 minute runs 20 samples, standards and controls = 24 hours or more
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