CIP Eco-innovation

Pilot and market replication projects Call 2009

Call Identifier: CIP-EIP-Eco-Innovation-2009

Interim Report BISCOL

Contract ECO/09/256112

Covering the reporting period from 01/09/2010 to 29/02/2012

Reporting Date 29/03/2012

Project coordinator: Rebecca Pogni Project website: http://www.biscol.unisi.it/

1 Progress of work plan in the period

1.1 General progress

The objectives of BISCOL project is to propose a new bio-dyeing process, based on the ecological optimisation of the overall dyeing process cycle. All the tasks foreseen for the first 18 months of the project have been fulfilled regarding textile pretreatments, synthesis of biodyes and new auxiliaries. The main deviation has been represented by the installation of the bioreactor for the scale up of the bio-dyes. All these aspects will be considered and analysed in details for each WP in the following part. Concerning the financial aspects, at month 18 more than 100% of expenses have been charged by partners with a mid term period financial situation in line with the Agency expectations.

1.2 Progress on all work packages against initial objectives

WP1 – The Project Management Guide has been produced at first month of the project with a deep description of all templates that will be filled by BISCOL beneficiaries in order to help the coordinator in reporting activities. All templates have been given at M1 and they can be downloaded by private zone of web-site. The management activity has been continued for all the reporting period (coordinating the work to be done in different WPs, updating the web-site and preparing and delivering all the documents requested by EACI). Different meetings has been organized: kick-off meeting, periodical and annual meetings of the whole consortium. Face to face meetings with different partners have been organized at any time was needed to plan and discuss research and financial activities. The Interim Report meeting has been organized in Bruxelles and the next project meeting will be held in Istanbul the 9th of July organized by SETAS.

WP2 – The WP has been closed at month 8 and all the deliverables have been supplied with the previous Progress Report. WP3 – Concerning the synthesis of new auxiliaries with low environmental impact all the tasks have been fulfilled both for wool and cotton fabrics and no deviations are foreseen. ACHIMO has investigated and produced some new formulated to be used for the dyeing of cotton fibres focusing their attention on their environmental impact, as it was done for wool fibres. Two different chemicals have been produced so far (Table 1): a surfactant (it is responsible for the control of the adsorption of the dyestuff onto the fibres) and a wetting agent (it is responsible for the penetration of the dyestuff into the fibres). The auxiliaries set-up by ACHIMO is a derivative of Pyrrolidone whilst the wetting agent is an ethoxylated compounds derived from natural sources. Both chemicals have been characterised in order to identify their environmental impact.

Table 1 – Auxiliaries developed by ACHIMO Component Biodegradability

(OECD 301F)

Toxicity

LC50 (Daphnia 96h)

COD

mg/kg

Surfactant 1-10 mg/L < 700

Wetting agent 28 d > 60% N.A. N.A.

Conventional surfactant (Amine Ethoxylated

quaternized)

28 d < 60% 1000 mg/L 1000

The new formulate has then been tested for the dyeing of cotton yarn with a conventional direct dyestuff (Blue Dilamine Luce – C.I. Blue direct 71) and the optimal concentration of the new auxiliary has been defined: 0.3 g/100 g cotton fibres it is required for the new product whilst for the conventional one it is needed 0.5 g/100 g cotton fibre.

On the same time, ACHIMO in collaboration with KING has carried out their activities related with the investigation of the performances of the Auxiliaries set-up for wool fibres with the Biodyes synthesized and delivered by UNISI. In particular the following Bio-dyes (one with a phenoxazine structure developed in the SOPHIED project and the other one with and azo bond) have been selected as promising for further investigations:

- CURIE 22: particular attention must be paid on the granulometry and in the storage. A new quantity has been synthesized by UNISI and supplied to ACHIMO for dyeing tests

- A red dye has been synthesized starting from diamino benzene sulphonic acid (DABS) at pH = 3

New precursors have been tested to explore the possibility to obtain a tri-color set (yellow, red and blue) to have access to the whole colour palette for woollen fabrics. With this in mind, the synthesis of new Bio-dyes from natural and chemical precursors has been investigated by UNISI on the basis of several chemical structure of known chemical dyes. In particular the following Bio-dyes has been synthetized by the small-scale pilot plant by WET (laccase immobilised in glass beads; sodium acetate and phosphate buffer 1mM; 25°C):

- Red dyes from 2,5-diaminobenzensulfonic acid, Acid Blue 62 and 2,5-diaminobenzensulfonic acid, 2,5-diaminobenzenesulfonic acid and 6-amino-4-hydroxy-2-naphthalenesulfonic acid (all dyes have been investigated at pH 3)

- Blue dyes from 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) and 4-(dimethylamino)-benzoic acid (investigated at pH 3), 3-methyl-2-benzothiazolinone hydrazone and 4-(dimethylamino)-benzoic acid (investigated at pH 6.5)

- Yellow dye from 3-amino-4-hydroxybenzensulfonic acid (CURIE 22; investigated at pH 4.5), catechin (investigated at pH 4.5), 3-Hydroxyanthranilic acid and 3-amino-4-hydroxybenzensulfonic acid (investigated at pH 3).

Some of them are under investigation by ACHIMO to test the dyeing properties on woollen fabrics. The optimisation of the reaction parameters and chemical structure will be performed for the most promising dyes in the next project period. All the dyestuffs synthesized so far are able to dye woollen fabrics but not cellulosic fibers like cotton. So different strategies will be attempted to obtain suitable dyes for cotton. The acid dyes for cotton have a chemical structure very similar to that of dyes for wool but it is crucial for cotton the length of the dye.

In this context, from one hand, UNISI will try to synthesize dyes with polymeric structure and SETAS will explore the possibility for chemical functionalization of CURIE 22 with reactive groups for dyeing of cellulosic fibers.

UNISI has also tested different natural precursors (such as salycilic acid, tyramine, gallic acid, acetosyringone, vanillic acid, vanillin, p-coumaric acid, syringic acid and syringaldehyde) and chemical precursors (such as sulfanilic acid, 1,4-diaminoanthraquinone, p-toluenesulfonic acid monohydrate, p-xylene-2-sulfonic acid hydrate, 2,7-dihydroxynaphtol, 1-hydroxybenzotriazole, caffeic acid, 1,2-dihydroxybenzene, 5-amino-1-(4-methoxyphenyl-1H-pyrazole-4-carboxylic acid, 5-amino-1-naphthalenesulfonic acid). These precursors have been tested by homocoupling and heterocoupling reactions. Furthermore, different route of synthesis using laccase are under investigation. Laccase activity has been tested in organic environment (such as ethyl acetate) using a natural substrate, ferulic acid. A yellow disperse dye has been produced with a yield of 42% and is under investigation for dyeing of synthetic fibres like polyester. WET has developed the cartridges containing immobilised enzyme. WET has worked on the set up of the enzyme cartridges to ensure larger productivity, piping and connection systems and optimal aeration systems. Concerning the bioreactor the main critical point to be addressed are the engineering and control of the bioreactor, together with the downstream processing as we are working in lower concentration than traditional chemical process. This has determined a delay in the installation of the bioreactor in SETAS due also to the difficulty in finding a third company for the production of the equipment.

WP4 – In the previous reporting period, NTT has optimized the pre-treatment under vacuum plasma of 98% woollen fabric. The study carried out on small scale showed that the nitrogen plasma is able to efficiently modify wool fibres by removing the fatty-layer responsible for the reduction of the water absorption kinetic. Thanks to the plasma pre-treatment, samples adsorb more dyestuff (model acid dyestuff Blue marino Dylan SB, C.I 284, 1% w/w wool fabric was used) and the combined use of plasma pre-treatment and auxiliaries (in WP4 the auxiliaries studied and produced by ACHIMO within the project were used at KING) allows to reduce the energy consumption of dyeing process: process temperature can be reduced up to 20 °C. The dyeing temperatures optimised with auxiliaries are shown in the figures below (Figure 1 and Figure 2).

Figure 1 – Reduction of the dyeing temperature induced by nitrogen plasma pre-treatment.

According to the preliminary results achieved with the 98% woollen fabric, NTT has investigated the efficiency of plasma treatment on the adsorption behaviour in function of the amount of wool fibre (from 75% to 100%, since these are the typical amount of wool in woolen fabrics) in the wollen textile. The analysis shows that the treatment is not affected by wool content.

Since the results on small scale were promising, NTT has made some tests with industrial machinery (the equipment has been set up and constructed by NTT) under atmospheric conditions to demonstrate that the process can be scale-up on production chain. A mixture of He (25%) and N2 (27%), an applied power of 565 W and a treatment speed of 8 m/min, were used as optimal process parameters. By applying these conditions in combination with the Auxiliaries produced by ACHIMO it is possible to reduce the dyeing temperature up to 12 °C (Figure 2) with a total reduction on energy consumption of 63 kJ/Kg of textile.

Figure 2 - Reduction of the dyeing temperature induced by atmospheric plasma pre-treatment.

At the same time, the analysis under vacuum treatment of cotton fibers has started. Preliminary results show that no significant changes on dyestuff adsorption of direct dyestuff are observed, even if the oxidation of the surface is observed: the yellow tendency of the treated sample is indicating the evolution of aldehyde groups on the cotton fibers. By replacing direct dyestuff with reactive dyestuff it is possible to achieve an increase in the dyeing yield with oxygen and nitrogen. In fact, the oxidation of the surface is able to increase the amount of functional groups (hydroxyl groups, amino groups or carboxylic groups) that can covalently react with the reactive dyestuff. No deviations are foreseen for this WP.

WP6 – The life cycle analysis carried out is focused on the auxiliaries synthesis and pre-treatment processes of the textile dyeing process of woollen fabric, both for the conventional and the BISCOL technologies (Figure 3). The assessment of these steps of the process allowed to understand the environmental burdens associated with the preliminary treatment of textile, in particular the use of innovative auxiliaries and plasma technology in comparison with the use of classic auxiliary (BASOLAN) associated to traditional pre-treatment process for epicuticle removal.

The analysis was performed taking into account all the processes constituting the dyeing process for the case of conventional dyes use. The data employed for auxiliaries and pre-treatment are set up for the industrial exploitation of the process.

System Definition and Life Cycle Inventory

BiscolBiscol ProcessProcessConventionalConventional ProcessProcess

PrePre--treatmenttreatment

DyeingDyeing

DyesDyes SynthesisSynthesis

AuxiliariesAuxiliaries ProductionProductionWaterWaterAcidAcid

Plasma Plasma PrePre‐‐treatmenttreatment

New New auxiliaryauxiliaryTraditionalTraditional DyestuffDyestuff

EnergyEnergy……

WaterWaterAcidAcid

TraditionalTraditional PrePre‐‐treatmenttreatment

TraditionalTraditional auxiliaryauxiliaryTraditionalTraditional DyestuffDyestuff

EnergyEnergy……..

INPUTINPUT INPUTINPUT OUTPUTOUTPUTOUTPUTOUTPUT

CODCODBODBODCO2CO2……..

CODCODBODBODCO2CO2……..

S1S1

S2S2

S3S3

S4S4

S5S5

S1S1

S2S2

S3S3

S4S4

S5S5

1 kg1 kgWollenWollen fabricfabric

1 kg1 kgWollenWollen fabricfabric

1 kg1 kgColoured Coloured fabricfabric

1 kg1 kgColoured Coloured fabricfabric

PurgePurge

WashingWashing

DryingDrying

Figure 3 – System definition and Life Cycle Inventory for Life Cycle Assessment for the conventional and the BISCOL technologies.

Calculations showed a better environmental performance for methods proposed by BISCOL. In fact, results confirmed that the relative impact of the pre-treatment process accounts for 73.4 % of the whole environmental impact of the traditional system, while the relative impact of plasma process accounts for about 3.8 % of the whole environmental impact of BISCOL system. The characterization step of the analysis allowed to calculate the relative contribution of results to all the impact categories. BISCOL process performances are better for all the environmental issues considered in the study as it can be seen in the following diagram (Figure 4).

Figure 4 – Characterisation step for pre-treatment conventional process and BISCOL technologies.

In particular, for the fossil depletion impact category, results attest a reduction of equivalent oil kilograms per kilogram of treated textile of about 67% and 71% for auxiliary 1 and 2, respectively. The energy saving is due to the acceleration of process time and to reduction of temperature required for textile substrate dyeing. For the water depletion issue, results show a reduction of cubic meters of water consumed per kg of treated textile of about 58% for both auxiliaries. To express the total environmental burden in a single score (Figure 5), we performed the calculation of the ecoindicators of the three analysed processes obtaining the lowest values for the Biscol pre-treatment processes (0.819 Pt for the conventional pre-treatment process, 0.231 Pt and 0.226 Pt for Biscol pre-treatment process with auxiliary 1 and 2, respectively).

Figure 5 – Environmental burden expressed as a single score of the three analysed processes: lowest values are obtained for the BISCOL pre-treatment processes. WP7 – The WP7 is focused on exploitation and business plan analysis. Preliminary results on business plan on bio-dye synthesis based on evaluation of laboratory costs (normalized considering the industrial scale up), have shown a tentative sale price for bio-dyes. A market analysis in Prato district has also shown an high interest regarding these new products. The sale price will be better defined at the moment that the semi-industrial bioreactor for dye synthesis will be operating. Atmospheric Plasma is a promising technology that could be easily transferred in the woollen fabric production line. A cost benefit analysis will be carried out in the second half of the project.

WP8 – The WP8 is focused on dissemination activity. Results concerning the first 18 months period of the project have been presented to different meetings/conferences and will be the objective of scientific papers. All the abstracts and presentations have been uploaded on our website for public consultation. A list of peer-reviewed and magazines articles, abstracts and oral contribution are reported following:

Peer-reviewed paper:

Martorana A., Bernini C., Valensin D., Sinicropi A., Pogni R., Basosi R. and Baratto M.C. (2011). “Insights into the homocoupling reaction of 4-methylamino benzoic acid mediated by Trametes versicolor laccase”. Molecular BioSystems 7, 2967-2969.

Magazine articles:

Techno Fashion 2012, vol. 34, 34-35. Title of article: “Sviluppi ecologici della nobilitazione: il progetto Biscol”. Author: Aurora Magni.

Meeting oral contributions: -Eko-Inovasyon Proje Teklif Cagrisi, 27th May 2011, Istanbul (Turkey). Title of contribute: “BISCOL Projesi”. Author: Rezzan Karaaslan. - Entrepreneurship, Values and Development: A Global Agenda, 3-6 December 2011, Istanbul (Turkey). Title of contribute: “Eco-Entrepreneurship–BISCOL”. Author: Rezzan Karaaslan. - VIII Ed. International School “Chemistry for Life and Environment”, University of Siena – Emory University (Atlanta-USA), June-July 2011Fungal laccases as tools for the synthesis of new hybrid molecules and biomaterials: Blue Enzymes for Green Chemistry”. Author: Rebecca Pogni -Open Day Technology Brokerage Event, 2nd December 2011, Next Technology Tecnotessile. Title of contribute: “The natural dye for industrial use – BISCOL”. Author: Rebecca Pogni

Abstracts: - XXXIX National Congress of Physical Chemistry, 20-24th September 2010, Stresa (Italy). Title of contribute: “Spectroscopic characterization of 4-(methylamino)-benzoic acid, a small precursor for the synthesis of new dyes”. Authors: Baratto M.C., Martorana A., Bernini C., Valensin D., Sinicropi A., Pogni R. and Basosi R. - X National Congress GIRSE, 26-29th September 2010, Rimini (Italy). Title of contribute: “Spectroscopic characterization of catalytic cycle in laccase mediator/precursor system”. Authors: A. Martorana, M.C. Baratto, A. Sinicropi, C. Bernini, R. Pogni, R. Basosi. - VIII Ed. International School “Chemistry for Life and Environment”, University of Siena – Emory University (Atlanta-USA), June-July 2011. Title of contribute: “Characterisation of Laccase-Catalysed Homocoupling Reaction of 4-(methylamino)-benzoic acid”. Authors: Baratto M.C., Martorana A., Bernini C., Valensin D., Sinicropi A., Pogni R. and Basosi R. - XXIV National Congress of Italian Chemical Society, 11-16th September 2011, Lecce (Italy). Title of contribute: “Insights on the homocoupling reaction of 4-methylamino benzoic acid mediated by Trametes versicolor laccase”. Authors: Baratto M.C., Martorana A., Bernini C., Valensin D., Sinicropi A., Pogni R. and Basosi R.

Del. N°

Deliverable name

Type WPN°

Delivery date from Annex I

Delivered (yes/no) and status (draft/final)

Submission with report

Forecasted delivery date

Comments on progress

1.1 Project Management

Manual 1 M1 Yes, Final PR1 M1 Done

Guide

1.2 Progress, mid term and final project reports to EACI

Technical description

1 M3,8,18,29,36 Yes, Final PR1,IR M3,8,18,29,36 Done

1.3 Minutes of project meetings to be sent also to EACI

Work organisation

1 M1,8,14,1823,29,36 Yes, Final PR1,IR M1,8,14,1823,29,36 Done

2.1 Report on the quantification and composition of the most used textile support

Technical description

2 M4 Yes, Final PR1 M4 Done

2.2 Report on chemical composition of chemicals applied in the textile processes

Technical description

2 M6 Yes, Final PR1 M6 Done

7.3 Business plan on synthesis of bio-dyes

Business plan

7 M12,36 Yes, Final M12 M12,36 Done

7.4 Business plan of dyeing process

Business plan

7 M12,36 Yes, Final M12 M12,36 Done

7.5 Monitoring protocol

Manual 7 M1 Yes, Final PR1 M1 Done

7.6 Reporting on monitoring

Technical description

7 M18,36,58 Yes, Final IR M18,36,58 Done

8.1 Project information updates

Text, ppt 8 M12,24,36 Yes, Final M12 M12,24,36 Done

8.6 Web-site realisation

Web site 8 M3 Yes, Final PR1 M3 Done

1.3 Identified deviations, problems and corrective actions taken in the period

WP3 - More effort then expected has been charged to WP3 by the partners involved in the synthesis of the Biodyes. In particular: WET: Wetlands has allocated more then twice, the hours foreseen at the beginning of the project. This is maily due to:

- Some skilled collaborators have left the company and so we have been obliged to use alternative collaborators with lower expertise but with lower salaries.

- The difficulties we had to face in developing the pilot for biotransformation. So this development took finally much more effort than planned.

This situation will not be a problem for the remaining part of the project as we have a budget left in line with the expectation. The main deviation from the objectives of the project has represented by the delay in the installation of the bioreactor in SETAS. On this context different corrective actions have been undertaken:

- The optimisation of the bio-dyes production has been performed in UNISI where a pilot is installed.

- a third company to build the machine has been identified and the invoice has just been sent to SETAS. The acceptance of invoice by SETAS is supplied in Annex 1. WET has agreed to follow the installation and supply the cartridges as soon as possible in the following period.

UNISI at M18 has almost allocated all the foreseen hours. This is mainly related with the larger effort put by UNISI in the optimisation of the biosynthesis of the new dyestuff. Moreover, the delay in the delivery of the pilot to SETAS has increased the involvement of its personnel in WP3 since they were uncharged to provide the dyestuff to other partner (in particular ACHIMO). On the contrary, the partners and in particular UNISI is underspending for the consumables (UNISI has spent less then 10% of the foreseen budget). WP4 - No deviations have been incurred during the period for WP4 from the technical point of view. Some deviations have been incurred on the resources to be allocated within the WP. In fact, WET and SETAS has not yet started their activities on plasma pre-treated samples. This is mainly due to the fact that WET is mainly involved in the optimisation of the bioprocessing (WP3) and to the delay in the delivery of the pilot plant to SETAS. Furthermore, NTT is underspending in the other costs category (NTT has to spend 15,000 € in the next ten months). WP6 - The efforts to be paid by final end-user was underestimated. A lot of data concerning the conventional production processes was required in order to perform a comprehensive study on the Impact of the new technologies and chemicals in the production line.

1.4 Progress regarding performance indicators

The Table 2 reported below, represents an overview on the performance indicators considering the first part of the process (plasma pre-treatment plus new auxiliaries for the

treatment of woollen fabrics) that has been analyzed so far using the LCA approach (Figure 4).

Table 2. Performance Indicators for BISCOL treatment of woollen fabrics

Objective Indicators Relative Impact Relative Impact Expected

CO2 ∼ 72% reduction 20% reduction Greenhouse gas emission

Methane1 68-70% reduction

15-18% reduction

Air quality Particulate matter ∼ 70% reduction 5% reduction

Mutagenic/carcinogenic Reduction /

Toxic (Human) ∼ 80%

Waste minimization 25% reduction (total wastewater)

15-20%

Improved Environmental Performance

Waste management

Hazardous waste 15-20%

Water Reduced water consumption

58%

5-7%

Better use of natural resources Energy Reduced energy

consumption 67-71%

15-18%

Patents

Market size in million Euros

Market potential

Market size in number of customers

Entry in new transational markets

Entry into different sectors

Economic Performance / Market Replication

Reduction of

cost per unit or process

1 According to the Recipe method used for the analysis the contribute to fossil depletion impact category has been computed as Kg of oil equivalent

2 Progress regarding market uptake and exploitation According to the results achieved at the Mid-term it has been demonstrated that Atmospheric Plasma is a promising technology that could be easily transferred in the woollen fabric production line. In fact, the capital cost investment is not so high (the cost of the pilot already available at NTT is around 65,000 € and a cost of around 100,000 € is expected for the large scale machinery) so a quick transfer is foreseen. According to the data collected for the LCA, an analysis of the benefit cost will be carried out in the second half of the project. All the auxiliaries and levelling agents set up by ACHIMO are ready to be transferred on the market.

3 Work plan for the next period

3.1 Planned activities in the next period WP3 - ACHIMO will optimise their products and their dosage on the dyeing recipe on the most promising Biodyes will be produced on pilot scale by SETAS. This activity will involve KING as well that must optimise the process conditions in terms of treatment time and temperature. UNISI will optimised the Bio-dye synthesis, new dyes from natural precursors, synthesis in biphasic system to obtain disperse dyes and synthesis of reactive dyes introducing specific functional groups by laccase. WET will proceed with the optimisation of cartridges and bioreactor performances. WP4 - As for the previous period, NTT will focus its activities in the optimisation and scale-up of the plasma process onto cotton fabrics. The effectiveness of the treatment will be investigated with both conventional and biosynthesised reactive dyestuffs will be selected by the consortium and/or produced within WP3. Since some of the biosynthesised dyestuffs have a similar structure to disperse one, in the case they could be efficiently exploited for the dyeing of Polyester, the effectiveness of the pre-treatment with this class of dyes will be investigated. WP5 – The dyeing parameters will be defined according to technical performance in terms of colour, brightness and colour resistances assessed by performing UV-Vis analysis and fastness tests. The effectiveness of the environmental friendly pre-treatments and of the reduction of the dyeing temperature will be evaluated by Life Cycle Assessment. WP6 - In next months, the research activity will be focused on life cycle assessment of the whole textile dyeing process of woollen and cotton fabrics, both via conventional and innovative eco-compatible technologies and taking into account the use of bio-synthesized dyes. Moreover, in order to complete the environmental analysis performed so far, a life cycle costing analysis will be integrated with the results already obtained on the pre-treatment steps of the process. The collaboration with partners of the Consortium will be of pivotal importance for data collection and the understanding of the main features of textile dyeing

processes. In turn, the study that will be performed in the next period will give fundamental feedbacks to the Consortium to define the strategy for the new proposed dyeing process.

3.2 Planned meetings, activities related to market uptake and dissemination activities.

The next project meetings will be held with the following programme:

- July 9th 2012 SETAS Turkey

- January 2013 KING Italy

- May 2013 SETAS Turkey

BISCOL project will be presented to the following meetings:

- Environmental Microbiology and Biotechnology 2012 (EMB 2012), 10-12th April 2012, Bologna (Italy). Title of the contribute: “Synthesis of textile dyes by laccase biotransformations”. Authors: Spinelli D., Martorana A., Baratto M.C., Basosi R. and Pogni R.

- Europhysics Conference on the Atomic and Molecular Physics of Ionized Gases 2012 (ESCAMPIG 2012), 10-14th July 2012, Portugal. Title of contribute: “Sustainable finishing of wollen fabrics by means of plasma treatment”. Authors: Fatarella E., Parisi M.L., Peruzzi F., Corsi L., Tacconi L., Basosi R. and Pogni R.

4 Overview on hours spent