NODES – No d O es Digi ale e Sos enibile
FLAGSHIP PROJECT GRIP
SPOKE 2 –GREEN TECHNOLOGIES AND SUSTAINABLE INDUSTRIES
DELIVERABLE D2.3.1: New da a abou he alo ized ma ices cha ac e iza ion and high
alue p oduc s
DELIVERABLE D2.3.2: New cha ac e ized ing edien s/ma e ials o ood, nu aceu icals,
pha ma and cosme ic applica ions
NB: D2.3.1 and D2.3.2 we e me ged
REPORTING PERIOD
Pe iod co e ed:
F om M 8 o M 30
Pe iodic epo da e and
e sion:
Ve _1
Deli e able # Responsible
UPO
RM2 Responsible
A lo io
This epo is pa o he p ojec NODES which has ecei ed unding om he MUR – M4C2 1.5
o PNRR unded by he Eu opean Union - Nex Gene a ionEU (G an ag eemen no.
ECS00000036)
1
Glossa y
Defini ion
Hub Coo dina o
(HC)
The Hub Coo dina o ep esen s he single poin o con ac o he
implemen a ion o he inno a ion ecosys em owa ds he MUR. I
ca ies ou he managemen and coo dina ion ac i i ies o he
inno a ion ecosys em, ecei es he undings, e ifies, and
ansmi s o he MUR he epo ing o he ac i i ies ca ied ou by
he Spoke and hei a filia es.
Na ional Reco e y
and Resilience
Plan (NRRP)
This documen uses he I alian ac onym o he NRRP, which is
PNRR (Piano Nazionale della Rip esa e Resilienza)
Resea ch P og am
Manage
The pe son who will be he esponsible o he o e all scien ific
con en s o he NODES p ojec . The NODES will appoin he
Resea ch P og am Manage . I e e s o “Responsabile del
P og amma di Rice ca” in he MUR’s Call o p oposal o
“Ecosis emi di Inno azione”
NODES’ Resea ch
and inno a ion
p og am
NODES’ Resea ch and Inno a ion p og am is a icula ed in
specific p og ams o each Spoke, wi h he aim o p omo e and
suppo applied esea ch on opics consis en wi h he In elligen
Specializa ion S a egy, wi h he guidelines o he 2021-2027
pa ne ship ag eemen scheme, wi h egional ope a ional plans
and egional and na ional esea ch and inno a ion p io i ies.
Al hough NODES’ Spokes a e concen a ed on di e en hemes,
hey will o ganize hei ac i i ies and ac ions wi hin a common
amewo k – NODES’ Boos e Me hodoloy
Spoke Coo dina o
The Uni e si y in cha ge o coo dina ing he Spoke’s ecosys em. I
e e s o “Spoke” in he MUR’s Call o p oposal o “Ecosis emi di
Inno azione”
Spoke Da a
Manage
The pe son who will be he esponsible o he moni o ing and
managemen o da a gene a ed a he Spoke le el. The Spoke
Coo dina o will appoin he Spoke Da a Manage .
Spoke Pa ne
The en i y associa ed o he Spoke Coo dina o . I can be an
Inno a ion Clus e , Compe ence Cen e , Resea ch Cen e ela ed
o he Spoke’s ecosys em and con ibu es o achie e objec i es
and impac unde he Spoke’ leade ship and managemen . I
e e s o “sogge i a filia i” in he MUR’s Call o p oposal o
“Ecosis emi di Inno azione”.
Spoke P ojec
manage
The pe son who will be he esponsible o he managemen ,
coo dina ion and p og ess o he p ojec a he Spoke le el. The
Spoke Coo dina o will appoin he Spoke P ojec Manage .
Spoke esea ch
and inno a ion
p og am
NODES’ Resea ch and Inno a ion p og am is a icula ed in
specific p og ams o each Spokes. The spoke will le e age a
consolida ed collabo a ion wi h leading p i a e and public
companies and will ocus he applied esea ch ac i i y on
echnological domains and applica ions ha can a ou he
in eg a ion o SMEs in o new alue chains.
Spoke Scien ific
and Technical
Manage
The pe son who will be he esponsible o he o e all scien ific
con en s o he p ojec a he Spoke le el. The Spoke Coo dina o
will appoin he Spoke Scien ific and Technical Manage .
2
Spoke
S akeholde s
Commi ee (SC)
Consul a ion s uc u e o med by ele an s akeholde s
(Go e nmen , uni e si ies, companies, ci il socie y, hi d sec o ,
e c.)
Spoke Thema ic
Gene al a ge ocus and domain o he Spoke esea ch.
Spoke Topics
Specific a eas/lines o de elopmen wi hin he Spoke.
Spoke Wo k
Package Leade
A he Spoke le el, Wo k Packages (WPs) will be o ganized by WP
leade s, who will be esponsible o pe o mance e alua ion and
epo ing.
Flagship P ojec
Main esea ch p ojec a he Spoke le el wi h he goal o
p o o yping, es ing, demons a ing he esea ch ac i i ies
owa ds highe TRLs.
3
Lis o Deli e ables
D. No
Name
Type
Dissemina i
on Le el
Due Da e
Deli e y Da e
(ac ual)
[numbe ]
[name]
[R — Documen ,
epo ] [DEM —
Demons a o , pilo ,
p o o ype]
[DEC — Websi es,
pa en filings,
ideos, e c]
[DATA — da a se s,
mic oda a, e c]
[DMP — Da a
Managemen Plan]
[ETHICS] [SECURITY]
[OTHER]
[PU — Public]
[SEN —
Sensi i e]
[R]
[C]
[S]
[mon h
numbe ]
[dd/mm/yyyy]
D2.1
Da a abou chemical
and nu i ional p ofiling
o aw was es and by
p oduc s
R
PU
M14
18/04/2024
D2.2.1
On line lab-scale
echnical mul i-de ice
o was e p ocessing*
O he
(ma e ial)
PU
M18
25/10/2024
D2.2.2
Deli e y o p o ocols
(lab-scale) eady o
scale-up he p oduc ion
o new high- alue
ma e ial (in
collabo a ion wi h
Companies)
R
PU
M22
16/02/2025
D2.3.1
New da a abou
alo ized ma ices
cha ac e iza ion and
high alue p oduc s
R
PU
M28
30/09/2025
D2.3.2
New cha ac e ized
ing edien s/ma e ials
o ood, nu aceu icals,
pha ma and cosme ic
applica ions
O he (ma e ial)
PU
M30
30/09/2025
D2.4.1
New da a abou
sus ainable p oduc ion
o new ma e ials om
biomasses
R
PU
M24
D2.4.2
New cha ac e ized
ma e ials om
biomasses
O he (ma e ial)
PU
M30
30/09/2025
D2.5.2
Po ous so ben s om
biomasses alo iza ion
O he (ma e ial)
PU
M30
29/09/2025
D2.6.1
New ma e ial p oduced
a pilo scale eady o
he o mula ion o
co- o mula ion (in
collabo a ion wi h
Companies)
O he (ma e ial)
PU
M32
D2.6.2
New well cha ac e ized
p ocess and o mula ed
pilo -p oduc s (in
collabo a ion wi h
Companies)
R/O he (Ma e ial)
PU
M32
4
D2.7
Repo on he
adso p ion and/o
ca aly ic pe o mances
o po ous solids de i ed
om biomasses
alo iza ion and
non- ecyclable plas ic
was es
R
PU
M32
29/09/2025
5
A) INTRODUCTION
The pa ne s in ol ed in he ac i i ies ela ed o he D2.3.1 and D2.3.2 (essen ially ela ed o
he ac i i ies lis ed in Sub-Task 2.2.2) wo ked wi h he aim o cha ac e ize he
ma e ials/ing edien s p epa ed a labo a o y scale, as epo ed in M2.3.1. The p incipal aim o
he ma e ial he e desc ibed is ela ed o he de elopmen o ing edien s eady o o mula e
ood supplemen s and cosme ic p oduc s, as well as o isola e bioac i e compounds o
pha maceu ical applica ions.
ROLE OF PARTNERS
The ole o pa ne s was dis ibu ed depending on i) he ma ix and ii) he acili ies
a ailabili y, bu pa icula ly ollowing he model p ocess selec ed as bes pe o ming
combined solu ions use ul o ob ain sus ainable new ing edien s eady o be scaled-up and
used in nu aceu ical/cosmeceu ical/ ood o pha maceu ical p oduc s.
The Deli e able (he e conside ed as p e iously planned as unique Deli e able me ging D2.3.1
and D2.3.2) epo s he cha ac e iza ion o he p oduc s as ou comes o he p ocesses.
As p e iously epo ed in D2.2.2, he ou ma ices selec ed we e 1. Ne ine oods 2. Rice by
p oduc s 3. F ui pomace and 4. Cow whey.
EXPLANATION OF THE WORK CARRIED OUT AND OVERVIEW OF THE PROGRESS
The o e all wo k o desc ibed he e was ela ed o he chemical/ unc ional cha ac e iza ion o
he ma e ials/ing edien s p oduced.
Following, he main ma e ials alo ized and cha ac e ized (and eady o be used o scale up
and o mula e new p oduc s, as expec ed) a e epo ed and desc ibed.
6
DSF-UPO
▪ Apple pomace ex ac
Fig. 1. Apple pomace a e p e-p ocessing (d ying, milling)
Apple pomace ex ac was ob ained h ough hea -assis ed ex ac ion (pa ame e s: 80 °C, 40
min, 1:10 ma ix : sol en a io) and subsequen eeze-d ying. The op imized pa ame e s o he
pilo p oduc ion we e op imized hanks o he use o ad anced ma hema ical modelling
(Random Fo es algo i hm, da a no shown).
The ex ac showed a significan con en o phlo idzin (a a ge o he wo k depending on he
bioac i e p ope ies o his compounds) when compa ed o he aw ma e ial, as well as a
significan ly high an ioxidan ac i i y. This p oduc can be used bo h in ood/nu aceu ical a ea
and cosme ic a ea.
Mo e p ecisely, he apple pomace was p e-p ocessed d ying he ma e ial a 40 °C o e nigh and
hen g ounding i using a bead mill, in o de o ob ain an homogeneous ma e ial (depending on
he a iabili y o he aw ma e ial, con aining also g ape seeds).
Apple pomace powde was hen s o ed a -20°C un il he analysis.
The mois u e con en o AP, be o e he d ying p ocess was 82.8 ± 0.4 % ( esh weigh ). The esul
ob ained is in ag eemen wi h wha has been epo ed in li e a u e. Mo eo e , on he esh aw
ma e ial, he lipid quan ifica ion was ob ained h ough a Soxhle appa a us, leading o ob ain
5.20 ± 1.16 % (w/w) o lipid. Has expec ed, his ela i e high alue was s ic ly ela ed o he
p esence o seeds in his by-p oduc .
The p oxima e composi ion is epo ed in Table 1.
P oxima e composi ion
Amoun
Mois u e % (d.w.)
12.3 ± 0.3
Ashes % (d.w.)
3.28 ± 0.03
P o ein % (d.w.)
4.32 ± 0.30
TDF % (d.w.)
33.5 ± 3.3
7
IDF
SDF
23.9 ± 1.6
9.53 ± 0.21
Table 1. P oxima e composi ion o apple pomace (AP): mois u e con en , ashes, p o ein and o al
die a y fibe (TDF) sum o insoluble die a y fibe (IDF) and soluble die a y fibe (SDF) con en .
Resul s a e exp essed as a e age ± s anda d de ia ion o h ee eplica es.
Rega ding he phenolic compound and he an ioxidan ac i i y, he analysis ob ained om he
ma e ial ex ac ed using e hanol/wa e (70:30, / ; solid/liquid a io o 1:10) showed esul in
ag eemen wi h li e a u e. To al phenolic con en (TPC) was 2.48 ± 0.11 mg/g (d.w.) exp essed as
ca echin equi alen , while an ioxidan capaci y (AC) measu ed wi h DPPH assay was 2.61 ± 0.01
mg/g (d.w.), exp essed as T olox equi alen . A p elimina y ex ac ion using only wa e as sol en
showed esul o 1.03 ± 0.07 mg/g o TPC and 2.05±0.02 mg/g T olox equi alen o AC (measu ed
wi h DPPH es ). This esul can be influenced by he p esen o suga s which a e co-ex ac ed
wi h wa e .
A e he op imiza ion o he phenolic’s ac ion ex ac ion (expe imen al design no epo ed
he e) he ex ac iche in phlo izin p esen ed an amoun o 50.3 µg/g, beside a significan
con en o hype oside and u in (e alua ed as a sum o que ce in glycosides) wi h an amoun o
95.2 µg/g. I was also possible o iden i y and quan ified cyanidin-3-o-galac oside (4.93 µg/g),
appa en ly he only an hocyanin p esen s in he sample. All hese cha ac e is ics allow o
conclude ha he p oduced ex ac is use ul bo h o nu aceu ical and cosme ic applica ions,
as well as eady – a e powde iza ion – o be included in ood supplemen s o mulae.
▪ Cocoa bean shells
Cocoa bean shells, as epo ed in p e ious Deli e able, we e p ocessed using di e en me hods
om di e en Uni s o RM2. The ollowing desc ip ion is ela ed o he p ocessing selec ed by
DSF-UPO.
Mo e p ecisely, cocoa bean shells we e i) di ec lyenzyma ically ea ed wi h Vyscozyme (1 %
w/w), ii) p e ea ed wi h ul asounds (15 minu es, 50 % ampli ude, 50% sonica ion cycle) and iii)
wi h a combina ion o bo h ea men s. Fig. 2 shows he aspec o he di e en fibe - and
polyphenols- ich ma e ials ob ained wi h di e en single o combined app oaches.
8
Fig. 2. Cocoa bean shells fibe s (be o e and a e ea men s)
Table 2 shows he di e en pa ame e s measu ed in he same p ocessed samples, pa icula ly
ocusing on fibe ac ion, polyphenolic ac ion, an ioxidan p ope ies and suga con en .
CBS
CBS+US
CBS+ENZ
CBS+US+ENZ
To al die a y
fibe (%)
~66,5%
(insolubile
~57,5%; solubile
~9,0%)
~48,4%
(insolubile
~40,3%; solubile
~8,2%)
~50,5%
(insolubile
~43,1%; solubile
~7,4%)
~49,4% (insolubile
~42,0%; solubile
~7,4%)
To al phenolic
con en (mg
GAE/g)
12,98 ± 0,17
5,43 ± 0,25
5,72 ± 0,13
5,85 ± 0,03
DPPH (mg TE/g,
azione solubile)
13,91 ± 0,25
9,28 ± 0,27
9,56 ± 0,17
9,59 ± 0,17
FRAP (mg TE/g,
azione solubile)
17,02 ± 0,30
11,01 ± 0,01
11,12 ± 0,07
11,52 ± 0,66
DPPH
QUENCHER (mg
TE/g)
2,59 ± 0,06
5,70 ± 0,31
10,57 ± 0,09
10,95 ± 1,31
To al
monome ic
suga s (mg/g)
1,68 ± 0,03
1,29 ± 0,04
8,53 ± 0,85
9,19 ± 0,09
Table 2. Composi ion o he cocoa bean samples p ocessed and eady o be used in model
nu aceu icals/cosme ics
The da a epo ed in he able 2 highligh significan di e ences be ween un ea ed CBS and
hose subjec ed o enzyma ic and/o ul asound ea men , as expec ed. Fi s , he alue o he
o al die a y fibe is significan ly highe in he un ea ed sample (~66% w/w), consis ing mainly o
insoluble fibe . The ea men s cause a ma ked educ ion in fibe con en : bo h he enzyme and
9
Fig. 6. Compa ison among he VOC’s p ofiles o AKO and PKO using HS-GC-IMS.
POLITO
The ac i i ies desc ibed in his epo we e ca ied ou in he amewo k o P ojec RM2,
Sub-Task 2.2.2, which aims o isola e high- alue ing edien s and compounds om was e
ma ices, wi h he objec i e o ob aining up-cycled ing edien s o applica ions in he ood,
nu aceu ical, and cosme ic sec o s. This ask builds on he esul s achie ed in Sub-Task 2.2.1,
mo ing om labo a o y-scale ex ac ions owa d pilo -scale p ocesses in o de o comple e
cha ac e iza ion and assess he easibili y o scaling up he mos p omising echnologies. In
pa icula , he alo iza ion o apple pomace and be y by-p oduc s has been conside ed o he
eco e y o polyphenols and pigmen s such as phlo e in and an hocyanins.
Du ing he las epo ing pe iod, di e en p e- ea men s and ex ac ion me hods we e applied
o imp o e he eco e y o o al polyphenols om apple pomace. The ac i i ies ocused on
he mal-s i ed ex ac ion (TSE), ul asound-assis ed ex ac ion (UAE), and mic owa e-assis ed
ex ac ion (MAE), all pe o med using wa e as he ex ac ion sol en . A kine ic s udy was ca ied
ou o de e mine he op imal ex ac ion condi ions, including solid- o-sol en a io, empe a u e,
ex ac ion ime, and applied powe . These op imal pa ame e s we e hen es ed a pilo scale,
and hei en i onmen al impac was e alua ed h ough li e cycle assessmen . Among he
in es iga ed me hods, mic owa e-assis ed ex ac ion p o ed o be he mos e ec i e, p o iding
he highes yields while also showing a o able en i onmen al pe o mance.
16
Ul asound-assis ed ex ac ion also ga e p omising esul s, and o his eason, i was conside ed
o scale-up.
In he ul asound ials, a fixed equency o 20 kHz was applied a cons an empe a u e (60 °C),
wi h a solid- o-liquid a io o 1:30 and an ex ac ion ime o fi e minu es. Scale-up expe imen s
we e conduc ed s epwise, mo ing om 50 mL o 500 mL, 1000 mL, and finally 5000 mL. The
fi s h ee olumes we e p ocessed using he Sonics Ma e ials VC 750 Ul asonic P ocesso ,
equipped wi h di e en p obes acco ding o he wo king olume, while he 5 L ial was ca ied
ou wi h he Hielsche UIP1000hdT-230 de ice (Fig.1).
Figu e 7. Apple pomace US p ocessing (Hielsche UIP1000hdT-230)
To al polyphenol con en was de e mined using he Folin–Ciocal eu me hod, and all
expe imen s we e pe o med in iplica e and s a is ically analyzed wi h one-way ANOVA and
Tukey es (Fig.
17
Tab. 7. To al polyphenol con en (mg GAE/g d y basis) ob ained a di e en ex ac ion olumes (50, 500,
1000, and 5000 mL). Da a a e exp essed as mean ± s anda d de ia ion (n = 3). Di e en le e s abo e he
ba s indica e s a is ically significan di e ences among ea men s a he 95% confidence le el (one-way
ANOVA, Tukey es ).
The esul s showed ha he polyphenol eco e y was compa able ac oss he ex ac ions a 50,
500, and 1000 mL, whe eas a significan dec ease in polyphenol concen a ion was obse ed a
he 5 L scale. This educ ion can likely be a ibu ed o se e al ac o s. Beyond he lack o
agi a ion and he limi ed pene a ion o ul asonic wa es a la ge olumes, he dissipa ion o
acous ic ene gy plays a c ucial ole: as he wo king olume inc eases, he ul asonic ene gy is
dis ibu ed ac oss a la ge mass, educing he in ensi y o ca i a ion in ce ain egions. Mo eo e ,
non-uni o m hea ing a highe olumes can gene a e localized ho o cold spo s; in ho spo s,
he mal deg ada ion o polyphenols may occu , while in colde egions he ex ac ion kine ics
may be educed. Ano he aspec conce ns he e ec i e su ace- o- olume a io, as pa icles in
suspension may sedimen wi hou s i ing, he eby educing sol en accessibili y. In addi ion,
he p opaga ion o ca i a ion is less e ficien in la ge olumes, wi h weake acous ic fields away
om he p obe ip.
Fu u e wo k will he e o e ocus on add essing hese limi a ions. New configu a ions wi h
imp o ed mixing and ene gy dis ibu ion will be es ed o enhance he homogenei y o he
p ocess a pilo scale. Adjus men s in ex ac ion ime and ene gy inpu will also be explo ed o
minimize deg ada ion phenomena while maximizing yield. Al e na i e ul asound equencies
o mul i-p obe sys ems may be conside ed o imp o e ca i a ion dis ibu ion in la ge olumes.
In pa allel, coupling ul asound wi h mild mechanical agi a ion o flow- h ough sys ems could
p o ide mo e ep oducible and scalable esul s. These e o s will con ibu e o he op imiza ion
o he p ocess, ensu ing ha he p omising esul s ob ained a labo a o y scale can be
ansla ed e ec i ely in o pilo -scale p oduc ion.
Final conside a ions and pe spec i es
The ac i i y he e desc ibed was p ima ily di ec ed o cha ac e ize he ma e ial upcycled and
alo ized (o ex ac ed/p ocessed) using “g een” me hods, as well as o comple e he
desc ip ion o he bes pe o ming ma e ial o be selec ed o he o mula ion o “final model
p oduc s” wi hin he pha maceu ical, nu aceu ical, cosme ic o fine chemical applica ions.
These p oduc s we e ob ained – also in close collabo a ion wi h some Companies in e es ed
o de elop new p ocesses and new p oduc s, as desc ibed in he P ojec - applying he bes
pe o med me hods/combined p ocesses selec ed a e he compa ison o 10 p ocesses (as
18
epo ed in D2.2.2).
The main ma e ial cha ac e ized (sou ce o bioac i e ing edien s) we e:
- Fibe om cocoa bean husks
- Ex ac ich in polyphenols and ca eine/ heob omine om cocoa bean husks
- Bioac i e polyphenols om apple pomace
- Bioac i e polyphenols and pigmen s om bluebe y pomace
- Oils om ui ke nels
Mo e ma e ials cha ac e ized in RM2 p ojec (mo e pa icula ly ice husks, soy, cow whey and
biomass om Elodea nu alii) a e desc ibed in D2.4.2.
19
