Biosensor for Food Quality

Food Quality

 Safety
 Shelf Life
 Consistency
Food Products as Analytical Samples

 Characterization of nutritional facts

 microbial content
 food additives
 pesticide and
 veterinary residues
What is biosensor

 Biosensors are analytical devices

incorporating a biological or biologically
derived sensing element either
integrated within or intimately
associated with a physico-chemical
transducer.
Biological recognition elements and
immobilisation strategies
 Three main classes of biological recognition
elements which are used in biosensor
applications
 enzymes,
 antibodies and
 nucleic acids
 bacteria
 single cell organisms
 whole tissues or higher organisms
Development of biosensors using
immobilised enzymes.
 Sensors have been used for the detection of toxic
compounds such as heavy metals, pesticides,
phenols, etc.
 Organophosphorous and Carbamates with
Acetylcholinesterase
 Heavy metals with urease
 Phenols with tyrosinase
 Because of different optimal operational conditions,
these enzymes cannot be immobilised on the same
sensor easily.
Antibody-based biosensors

 Antibodies may be polyclonal, monoclonal or

recombinant, depending on their selective
properties and the way they are synthesised.
 They are generally immobilised on a
substrate, which can be the detector surface,
its vicinity or a carrier
 The bio-molecule immobilisation step is critical in
the development of any sort of biosensor.
 The immobilised bio-molecule needs to keep its
original functionality as far as possible in order for
the biosensor to work.
 Recognition sites should not be steric hindered
 The type of transduction used,
 The nature and composition of the sample
 The possibility of multiple use of the biosensor
Optical biosensors

 These are probably the most popular in

bioanalysis, due to their selectivity and
sensitivity.
 Optical biosensors have been developed for
rapid detection of contaminants, toxins ,drugs
and even pathogen bacteria
Surface plasmon resonance

 SPR biosensors measure changes in

refractive index caused by structural
alterations in the vicinity of a thin film metal
surface
 SPR has successfully been applied to the
detection of pathogen bacteria by means of
immunoreactions
Drawbacks

 Complexity
 high cost of equipment
 large size of most currently available
instruments
Electrochemical biosensors

 These devices are mainly based on the observation

of current or potential changes due to interactions
occurring at the sensor sample matrix interface.
 Techniques are generally classified according to the
observed parameter: current
(amperometric),potential (potentiometric) or
impedance (impedimetric).
 It allows to work with turbid samples, and the capital
cost of equipment is much lower.
 electrochemical methods present are slightly more
limited in terms of selectivity and sensitivity
Amperometric methods
 It works on the grounds of an existing linear relationship between
analyte concentration and current.
 The sensor potential is set at a value where the analyte, directly
or indirectly, produces a current at the electrode.
 In the case of biosensors, where direct electron exchange
between the electrode and either the analyte or the biomolecule
is not permitted, redox mediators are required.
 Redox mediators are small size compounds able to reversibly
exchange electrons between both the sensor and the enzyme of
choice.
 Bacterial biosensors do not differ much from more conventional
biosensors
Potentiometric methods

 They consist of an ion selective membrane

and some bioactive material, e.g., an
enzyme.
 The enzyme catalysed reaction consumes or
generates a substance which is detected by
the ion-selective electrode.
 Since potentiometry yields a logarithmic
concentration response, the technique
enables the detection of extremely small
concentration changes.
Biosensors for Food
Contaminants Analysis
Microorganisms in Food

 specific microbiological and biochemical

identification.
 Nucleic acid based assays are effective
identification tools, though they are limited, in that
they will indicate the presence of a microorganism,
but do not give any information on effective toxins or
virulence.
 immunological techniques are used to detect
microbes by their unique antigenic determinants-
information on the presence of antigens regardless
of whether the microorganims are viable or not.
Label Free Methods

 Monitoring of analyte at the solid (i.e., sensor)

or liquid (analyte solution) interface.
 The basis of this detection mode is the
physical phenomena occurring on the
transducer’s surface during the biochemical
reactions (e.g., change in optical or electrical
features).
Bioluminescence Sensors

 enzymes emit photons as a product of

enzymatic reactions.
 introduction of the gene encoding for the
enzyme luciferase in the genome of a
bacteriophage.
 host bacterium is infected, the luciferase
encoding gene can be transferred to the
bacteria, conferring bioluminescence
Piezoelectric Sensors
 Principle:
 The binding of the analyte to receptor previously
immobilized onto the surface , increases the mass on the
sensor and determines a proportional decrease of the
oscillation frequency of the piezoelectric crystals.
 Piezoelectric sensor surfaces can be functionalized with
antibodies raised against the whole microbial cells,
surface expressed antigens, microbial toxins or even
DNA specific sequences.
 Staphylococcal Enterotoxin B, Candida albicans ,
Listeria monocytogenes
Piezoelectric Sensors
Indirect Detection of Bacteria

 the indirect biosensing of bacteria is based

either on the use of a labelled molecule or on
microbial metabolism tracking.
 viable and nonviable cells
 cell metabolism, responds to viable cell
concentrations
Antibody Based Biosensors

 Immuno-sensors
 immunochemical
Immunosensor for hemolytic organisms

 hemolytic bacteria to disrupt a lipid

membrane
 Antibodies- immobilized on the surface of a
platinum working electrode.
 liposomes containing an electrochemical
mediator were added to the medium.
 presence of hemolytic organisms determined
the rupture of the lipid bilayer of liposome,
releasing the electrochemical mediator in the
medium
DNA/RNA Based Biosensors

 high specificity of DNA hybridization with the

inherent sensitivity of electrochemical
transducer
Metabolism Based Biosensors

 redox reactions of the metabolic pathways

mediated via an electrochemical mediator.
 information on the viable cells
 poor selectivity
 transducer can either detect the consumption
of oxygen or of another electro active
metabolite.
Antibiotics in Food

 toxicity, allergic reaction and the development of

bacterial resistance.
 high performance liquid chromatography (HPLC),
gas chromatography mass spectroscopy (GC-MS)
and liquid chromatography mass spectroscopy (LC-
MS).
 Microbial inhibition assays, and immunoassays
 surface plasmon resonance-detection of
glycopeptides antibiotics in a fermentation process
Heavy Metals in Food

 atomic absorption spectroscopy, inductively

coupled plasma optical emission
spectrometry, and inductively coupled plasma
mass spectrometry (ICP-MS).