Workshop: The Milk Day, Trento 4th February 2015 - Symphony Project, www.symphony-project.eu 1

THINKSHOP

THE MILK DAYTRENTO, 4TH FEBRUARY 2015

In dairy industry, one of most pressing unmet needs is the

timely detection of toxins contaminations that originate

from animal feed and are secreted into milk.

The available technology for aflatoxin detection is

laboratory-based, in most cases requires sample

preparation and does not provide timely identification of

contaminants, thus fails to deliver cost-effective

management of milk quality.

Can the use of Micro System Technologies and biosensors

provide a solution to overcome these limitations?

What are the major factors that could jeopardize the

integration and automation of current analysis

techniques?

Milk chemistry and composition, strict regulations for milk

quality, dairy industry requirements, the large diffusion of

milk consumers and producers pose substantial

challenges, but, in the meanwhile, offer a number of

opportunities for exploiting the huge potential of

miniaturized systems and sensors.

This workshop, organized in the framework of the

SYMPHONY project, will be the occasion to discuss relevant

open issues in the application of MST in the analysis of

milk contaminations.

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Program

14.10- Introduction and presentation of SYMPHONY project

Andrea Adami, FBK-Technical manager of the SYMPHONY project

14.30- Milk quality testing and regulations

Ulrich Leist, DRRR, Germany

14.50- Milk and health in the Province of Trento

Alberto Mattivi, Prevention Dep. Local Health Trust, Trento, Italy

15.10- Milk quality testing in Trentino

Nicola Cologna, CONCAST, Trento

15.30- Rapid Detection of Aflatoxin M1 in Milk: Analytical Challenges and Validation Aspects under EC Perspective

Veronica Lattanzio ISPA-CNR, Italy

15.50- Research & Business - Transitioning Innovation to Commercial Success

Mark Whatton, QCL, United Kingdom

16.10- Coffee break

16.40- MST for food quality and safety

Andrea Adami, FBK-Technical manager of the SYMPHONY project

17.00 - Discussion

18.00- Conclusion

FP7-ICT-2013-10 Grant agreement no: 610580

SYMPHONYIntegrated SYsteM based on PHOtonic Microresonators and

Microfluidic Components for rapid detectioN of toxins in milk and diarY products

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The aflatoxin issue in milk industry

Aflatoxin M1 is a milk contaminant and potent carcinogenclassified in group 1 of the International Agency for theResearch on Cancer (IARC, 1993).

Aflatoxin B1 is produced by the fungus Aspergillus flavus andcan contaminate feedstock. Aflatoxin M1 is a metabolite ofaflatoxin B1 and is secreted into milk.

Thermally stable during pasteurization or UHT treatment

In the European Commission regulation (EC) No. 1881/2006, themaximum level of aflatoxin M1 contamination in milk is set to50 ppt and to 25 ppt for infant formulae, as a vulnerable groupof the population

Aflatoxin is screened periodically by ELISA or lateral flow kits,with HPLC used as certified method for confirmation of positivecases

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Recent cases in Italy

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The project at a glance

• The objective of the SYMPHONY project is the development of a system for thedetection of aflatoxin M1 in milk for the dairy industry, that can take samplesat various stages during milk production when milk is transferred betweenstorage sites.

• The SYMPHONY project will deliver and test in real settings a smartheterogeneous integrated system by the integration of key enablingtechnologies such as micro-nano-bio-systems (MNBS) and photonics,polymer-based technologies for low-cost microfluidics and surfacefunctionalisation.

• The vision is to develop an analytical system with sensing capabilities to detectaflatoxin M1 but also extensible also for multiple parameters detection.

Research Centers and University

• Fondazione Bruno Kessler, Trento (IT)

• ACREO Swedish ICT (S)

• Department Physics of University of Trento (IT)

Companies

• Lionix BV (NL)

• Epigem Ltd. (UK)

• CONCAST - Consorzio Dei Caseifici Sociali Trentini (IT)

• Quadrachem Laboratories Ltd. (UK)

7 partners - 4 Countries 3 (I), 2 (UK), 1 (NL), 1(S)

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The consortium

http://science.unitn.it/~semicon/

www.acreo.se

http://www.lionixbv.nl/

http://epigem.co.uk/

http://www.trentingrana.it

www.fbk.eu

http://www.qclscientific.com/

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Key Technologies

SYMPHONY project addresses scientific and technical advancement in thefollowing technical fields:

• Biofunctional surfaces for sensors and sample treatment, based on aptamersand antibodies, for the optimisation of sensitivity and limit of detection.

• Development of microfluidic modules for fat removal, aflatoxin extraction,pre-concentration and elution.

• Photonic devices, by the development of new devices based on Micro RingResonators (MRR) and asymmetric Mach-Zender interferometers (aMZI).

• Hybrid integration of photonic sensors in a monolithic photonic system,including VCSEL and photodiodes.

• Integrated heterogeneous systems, by the merging of photonic devices andmicrofluidics for automated systems in milk quality control.

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Milk Chain and Safety Testing at dairies

Compliments existing testing procedures

within dairy industry

Access to experienced technicians to operate and maintain system

Could immediately assist dairies with

meeting legislation

Test time needs to be within 5 minutes so tanker acceptance

times are not increased

Robust system required to survive typical milk

intake laboratory

Rapid uptake due to end user familiarisation with analytical systems

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Sample preparation by continuous flow microfluidics

Fat separation

• pre filtration may be needed to remove non-milk debris and milk "clots"

• Centrifugation replaced by continuous flow fat removal

• Numbering up or manifolded multiple devices provides increased throughput

Concentration

• Immunoaffinity concentration of aflatoxin to:

• Increase the limit of detection of the system

• Reduce the interference of milk matrix

• Release by temperature or pH shock

detection

• Microseal interface to sensor is required

• Sensor based on mRR or MZI with integrated VCSEL, detectors

• A valve controlled sub-system could be used for regeneration of sensor surface using valve control of a sensor flow cell linked to reservoirs on-or-off chip

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Integrated photonic system

The photonic system is based on:

• LioniX Si3N4/ SiO2 single stripe technology and FBK SiON technology

• Sensor architecture: MRR/MZI

• The integration strategy: butt end coupling

• Cross section and sensor overview (planar and vertical coupling)

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Integration of a compact photonic sensor system

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The challenges

What are the major challenges for microsystems for milk analysis?

1. Milk is a complex matrix, including multiple phases and a number of chemical species.

Casein micelles, typical size is 62-154nm, fat globules, also including proteins,

phospholipids, glycoproteins, neutral lipids, enzymes and other minor components.

Typical size is about 0.2-8µm, water-based fraction includes whey proteins, salts, sugars,

vitamins. Aflatoxin and device itself interact with the matrix.

2. Simple and fast systems are more likely to meet the requirements (cost and time for

measurement)

3. Robust systems (including calibrations and measurement repeatability)

4. Matching user practices