CORDIS - EU research results



Reporting period: 2016-07-01 to 2017-06-30

Sulphites are food additives with antioxidant and preservative properties. However, they are recorded as allergens by the main international regulatory bodies on food safety because of their adverse health effects. Hence, sulphites maximum concentration in foodstuff is regulated, and must be ensured by the agro-food processing industries. Current methods such as Monier Williams are not solving the problem of the high quality control standards required to comply with new regulation on food safety and consumer health protection. The agro-food industry demands new technology enabling fast, accurate and simple methods for sulphite level monitoring and traceability along the whole supply chain. BIOLAN has already developed at the laboratory scale a simple, reliable and specific enzymatic biosensor based on solid electrodes for sulphite determination.
Within the frame of the project SO2SAFE BIOLAN has developed a novel miniaturized ready to use biosensor based on disposable screen printed electrodes (SPE) which will allow a highly sensitive, selective, fast and user friendly method for determination of sulphites and has achieved a scale-up manufacturing capacity for cost-competitive production of the developed biosensors.
As result of this innovative development BIOLAN offers a complete solution for sulphite level monitoring and traceability solution (see attached figure) that comprises:
 BIOFISH 300 for crustacean analysis: is a benchtop device for laboratory use, calibration is needed before sample analysis. Once calibrated, several analyses can be performed, sample analysis takes 3 minutes.
 BIOFISH 700 for crustacean analysis: portable device, sample treatment is needed. Suitable to be installed in laboratories, industries, retails, raw material reception points.
 BIOFISH 700 for water analysis: portable device that requires minimal sample treatment. Suitable to be applied for field applications for early monitoring of water in crustaceans processing.
SO2SAFE biosensors consist of a simple use measuring device and the biotests targeted to all agro-food industries where sulphites are used as additives at different steps of the supply chain. The novel solution will be relevant not only for the food industry economy, but also for the society due to the direct impact that the sulphite has in health.
Main specifications for the development of a miniaturized biosensor for sulphite concentration determination in crustaceans and for scale up manufacturing process have been defined. Specifications for materials, processes, subcontractors and manufacturing conditions have served as initial guidelines for suitable biosensor development and manufacture.
The novel sulphite biosensors were constructed by modifying the working electrode of the SPEs with Sulphite oxidase (SOx) enzyme. Amount of each component, working potential and time analysis were optimized in order to achieve the highest sensitivity and stability.
The developed biosensors were applied to:
 the detection of sulphite in crustaceans in a linear range of 40 to 120 mg/kg of sulphite with a stability of 180 days obtaining good recovery rates for real samples previously analyzed by the official method, Monier Williams.
 the detection of sulphite in water in a linear range of 19 to 76 g/L showing a stability of more than 180 days. During demonstration activities in Ecuador and Honduras, water sample of the bins where shrimps are collected were analyzed and compared to the results obtain with the validated benchtop biosensor BIOFISH 300 obtaining equivalent results (10% error).
Validation activities were carried out for the SO2SAFE biosensors with water samples. The results showed that:
 The method is linear.
 Sodium sulphite, sodium metabisulphite, potassium metabisulphite and formaldehyde metabisulphite, positive interfering compounds, are also detected by the biosensor.
 Recoveries were in the range of 85 to 115% .
 SO2SAFE biosensor provided accurate and precise sulphite quantification.
 The LOW was established at 13,62 g/l and the LOD at 4,54 g/l.
A miniaturized measuring device, a portable potentiostat, was design and developed to read the disposable biosensors based on SPEs. Different tests were performed to demonstrate compliance with EU Directives regarding electromagnetic compatibility stating that SO2SAFE device complies with them and is suitable for CE marking.
Manufacturing layout of SO2SAFE biosensors is available and tested, a high manufacturing capacity is achieved for screen printed electrodes, enzyme and biofunctionalization and assembly process.
Exploitation of SO2SAFE results will be carried out by direct commercialization of SO2SAFE biosensors from BIOLAN international sales network and through further development of SO2SAFE technology platform into other food safety applications, such as the analysis of malic acid in grapes, musts and wines.
Several activities for demonstration and dissemination of SO2SAFE results were executed by BIOLAN targeted to the main stakeholders and the general audience. During SO2SAFE action BIOLAN attended to the main international trade fairs for the seafood industry in Europe, Asia and Latin America, demonstrated SO2SAFE biosensors performance in real aquaculture facilities of final customers, and launched a multi-media communication package including several videos, posters, and new corporate brand-image. The dissemination of project results was performed through live presentations in several international events launched by the EC, articles in main international journals, and as exhibitor in the main international aquaculture and seefood trade fairs.
Progress beyond the state of the art:
The table attached summarizes the main sulphite monitoring methods currently used and compares them with the miniaturized sulphite biosensor developed according to the most relevant criteria.
In 2016 BIOLAN launched to the market the bentchtop biosensor developed for sulphite quantification in crustaceans (BIOFISH 300). At the beginning of SO2SAFE project this device was at prototype stage. Crustacean industries are using this solution; the acceptance of this technology was very good. It is a very simple method but needs to be calibrated and its use is limited to the laboratory. Crustacean industries are demanding a complete solution for sulphite monitoring. SO2SAFE biosensor (BIOFISH 700) has critical advantages, precalibrated strips or biotests, less than 1 minute to obtain the result and portability for field application.

Potential impact:
Sulphite monitoring is still a priority for the crustacean industry. 53 cases of high concentration of sulphites in crustaceans were reported to the Rapid Alert for Food and Feed (RASFF) portal between July 2015 and June 2017. Most cases were related with contaminated shrimps imported from countries outside Europe, that reveals the importance of monitoring sulphite in the raw and processed material that comes from Europe and moreover from outside Europe.
BIOLAN’s sulphite quantification solution based on biosensors enables the combination of the accuracy of the verification methods such as Monier Williams with the rapidity and simplicity of on-site methodologies.The user-friendly solution developed by BIOLAN allows the monitorization of sulphite all across the processing chain of shrimp production reducing the risk of contamination with high sulphite concentration.
SO2SAFE biosensor progress beyond state of the art
SO2SAFE biosensor complete solution for sulphite analysis in seafood