HARMFUL ALGAL TOXINS DETECTION FOR REGULATORY …iocwestpac.org/files/upload_manual/Toxin...
Transcript of HARMFUL ALGAL TOXINS DETECTION FOR REGULATORY …iocwestpac.org/files/upload_manual/Toxin...
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HARMFUL ALGAL
TOXINS DETECTION
FOR REGULATORY
PURPOSE
GIRES USUP
Universiti Kebangsaan Malaysia
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Vilarino et al. (2013) Detection of aquatic algal toxins has become critical for the protection
of human health. During the last 5 years, techniques such as optical, electrochemical, and piezoelectric biosensors or fluorescent-microsphere-based assays have been developed for the detection of aquatic algal toxins, in addition to optimization of existing techniques, to achieve higher sensitivities, specificity, and speed or multidetection. New toxins have also been incorporated in the array of analytical and biological methods. The impact of the former innovation on this field is highlighted by recent changes in legal regulations, with liquid chromatography-mass spectrometry becoming the official reference method for marine lipophilic toxins and replacing the mouse bioassay in many countries. This review summarizes the large international
effort to provide routine testing laboratories with fast, sensitive, high-throughput, multitoxin, validated methods for the screening of seafood, algae, and water samples.
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TOPICS
Toxicities and toxins
Current status of detection methods
Issues and challenges
Should we ‘reinvent the wheel’?
How much are we willing to pay to keep our seafood
safe?
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Paralytic shellfish poisoning
and PSTs
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Regulatory status Regulated: Yes
Regulatory limit: 800 ug/kg STX eq (600 in some countries)
Current methods:
MBA with 0.1M HCl (15min) (EC) No 2074/2005
LOD: 370 ug/kg STX eq
HPLC-FLD (Lawrence method) (EC) No 1664/2006
LOQ: 10-80 ug/kg STX eq for individual analogs
RBA: LOQ in the femto-molar range STX eq
Standardized methods:
AOAC method 959.08 (MBA)
AOAC method 2005.06 (HPLC-FLD)
AOAC OMA-2011.27 (RBA)
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Potential MBA replacement
for PSTS
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Diarrheic shellfish poisoning and toxins
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Regulatory status
Regulated: Yes
Regulatory limit:
OA: 160 ug/kg OA eq
Pect-2: 160 ug/kg OA eq
AZA-1: 160 ug/kg AZA eq
YESSO: 1000 ug/kg YTX eq
Current methods:
MBA or Rat bioassay with acetone extraction (24h)
(EC) 2074/2005
Standardized methods:
None yet
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Potential MBA replacement for DSP toxins
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Amnesic Shellfish Poisoning
and toxins
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Regulatory status
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Ciguatera Fish Poisoning and Toxins
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Regulatory status
Note: EU and WHO suggested limit of 0.01 ug/kg P-CTX1 eq as safety limit for human
consumption
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Issues and Challenges Toxin standards
Equipment
Personnel
Transportation
Operational budget
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How much are we willing to
invest? Equipment
LC/MS/MS
Scintillation counter
Associated extraction consumables
Manpower
Training of dedicated personnel
Operational costs and sustainability
Possible cost-sharing between government and industry
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Anal Bioanal Chem. 2013 Sep;405(24):7719-32. doi: 10.1007/s00216-013-7108-6.
Epub 2013 Jul 3.
Innovative detection methods for aquatic algal toxins and their presence in the food
chain.
Vilariño N1, Louzao MC, Fraga M, Rodríguez LP, Botana LM.
Author information
Abstract
Detection of aquatic algal toxins has become critical for the protection of human
health. During the last 5 years, techniques such as optical, electrochemical, and
piezoelectric biosensors or fluorescent-microsphere-based assays have been
developed for the detection of aquatic algal toxins, in addition to optimization of
existing techniques, to achieve higher sensitivities, specificity, and speed or
multidetection. New toxins have also been incorporated in the array of analytical and
biological methods. The impact of the former innovation on this field is highlighted by
recent changes in legal regulations, with liquid chromatography-mass spectrometry
becoming the official reference method for marine lipophilic toxins and replacing the
mouse bioassay in many countries. This review summarizes the large international
effort to provide routine testing laboratories with fast, sensitive, high-throughput,
multitoxin, validated methods for the screening of seafood, algae, and water
samples.
PMID: 23820950 [PubMed - indexed for MEDLINE]
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Thank you
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Molecular Interactions
Interactomics
Signal Transduction
Cell Adhesion
Antibody Binding
Endogenous Receptor-Ligand
Interactions
DNA Binding Proteins
Drug – Target Interactions
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What can be studied?
Scope of Technique
Proteins
Nucleic Acids Lipids & membrane molecules
Carbohydrates
Small-Molecule Drugs
Whole Cells
Viruses Bacteria
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Biomolecular Interaction Analysis: BIAcore
Detection principle: Surface Plasmon Resonance: SPR
One binding partner (LIGAND) immobilised on chip
Other (ANALYTE) injected: microfluidics
PC collects binding data in real time
Chip is regenerated to remove analyte
Cycle is repeated
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Comprehensive information
Molecular interactions in real time:
Detect
Yes/No
Identify
Specificity
Binding partners
Characterize
Affinity
Kinetics
Epitope mapping
Concentration
Thermodynamics
Mass Spec Link-Up
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Versatility
• Range of chip surfaces
• Range of coupling chemistries for different needs
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Surface Plasmon Resonance
• Critical angle of polarised light total internal reflection
• Results in excitation of surface plasmons in the gold
• Creates evanescent wave field dissipates into sample matrix
• Intensity of reflected light depletes
• Critical angle changes with changes in sample matrix (binding)
• Change in angle is converted to resonance signal directly proportional to mass
bound at surface