AM0209_Analytical Methods for the Determination of Mycotoxins in Food_Widiastuti Setyaningsih

8/8/2019 AM0209_Analytical Methods for the Determination of Mycotoxins in Food_Widiastuti Setyaningsih

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Michael Z. Zheng, John L. Richard and Johann Binder

Presented byWidiastuti Setyaningsih

Under Supervision byProfessor Armando

Durant

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INTRODUCTIONMycotoxin Producer

Contamination

FOODBORNE

DESEASE

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Mycotoxin AnalysisConventionalAnalysis Rapid Method

Analysis

Easier to useFasterLess expensive

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Rapid Methods for MycotoxinAnalysis

The term ‘rapid method’ usually refers to a method muchfaster than respective reference methods and also has atendency of promoting the method.

1. ELISA2. Membrane based immunoassay

Flow-through assay

Lateral ow test1. Flurometric AssayImmunoaffinity column clean-upSPE column clean-up

1. Fluorescent polarization

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Antibody

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Enzyme linked immuno-sorbent

assay (ELISA)(a)Sample mixedwith conjugate;

(b)Mixed contentadded to antibodycoated well;

(c)Mycotoxin bindsto antibody in the1st incubation;

(d)Unboundmaterials arerinsed away in thewashing step;

(e)Substrate isadded to developcolor;

(f) Stop solution isadded to stop the

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• Direct Competitive ELISA• Anti-mycotoxin anti-body is coated on a membranesurface• Mycotoxin and mycotoxin- enzyme conjugatecompete for the

limited antibody binding sites.

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Membranebased

immunoassay: Flow-throughassay

Principle of membrane-basedow through test• (in the absence of toxin (left),

color is developed;• in the presence of toxin (right),

color development is suppressed).

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A typical immunochromatography test strip• Sample pad,• Conjugate pad,• A membrane,• An absorbent pad and• An adhesive backing.

The competitive reaction scheme is used mostoften when testing for small molecules withsingle antigenic determinants such asmycotoxins

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Membranebasedimmunoassay :Lateral ow test

Schematic illustrationof a lateral ow test.(a) Conguration;(b) Reagents placement.

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contains anti-mycotoxinantibody that isimmobilized ontoa solid supportsuch as agarose gelin phosphate buffer,

all of which is containedin a small plasticcartridge

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Principle of immunoaffinitycolumns

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Solid-phase extraction (SPE)column clean-upSPE columns or cartridges are

Silica gel,C18 bonded to silica gel,Forisil or ion exchange resins.

A one-step SPE cleanup

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Solid-phase extraction (SPE)column clean-up

Principle of a one-step solid phase extraction

column

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1. Evanescent wave technology2. MIPs3. Microarray technology4. Luminex xMAPÒ technology

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Evanescent wave technologySurface plasmon resonance

biosensorPrinciple of Surface PlasmonResonance (SPR)L: light source,D: photodiode array detector,P: prism,S: sensor surface,F: ow cell.

(The two thick lines in the reected

beamprojected on to the D symbolize thedropping of light intensity followingthe resonance phenomenon at time =t1 and t2. The line projected at t1corresponds to the situation before

binding of antigens to the antibodieson the surface and t2 is the position

b

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Fiber OpticImmunosensor

Principle of Fibre Optic Immunosensor

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Critical ViewInsufficient validation of methods causes the methods to belimited to those matrices for which they were validated

(ELISA) methods also need to be validated and developed thatwill distinguish among the mycotoxins individually orcollectively ( not only valid for each the matrix ) since thecross reactivity might occur.

Even the method is quantitative or semi-quantitative,confirmation analysis still need to be conducted.

The microarray technology can be used not only for microbialfungus determination as mentioned by the author, but also forthe simultaneous detection of mycotoxin (aflatoxin B1 and

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Critical ViewNo conclusion that may mention the recommended rapidmethod which is considered has better performance formycotoxin analysis and apart from MIPs, the author did not

describe how the methods have the possibility as potentialtechnologies to be able to apply for mycotoxin analysis

The fluorometric assay with SPE clean-up cannot becomparable since the analyte only aflatoxin B1 instead of totalaflatoxin (B1, B2 and G1).

This review may helpful for the food producers or food inspector institutions to choose the suitable method tomonitor the quality of the product apart from mycotoxincontamination

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