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Transcript of Challenges in co-extraction of marine algal active ...nordicinnovation.org/PageFiles/9098/Franck...
Challenges in co-extraction of marine algal active ingredients:An exemple of co-extraction of alginates and fucoidans
Franck Hennequart,Director Research & Innovation,ALGAIA St Lô
Algaia™ portfolio
Red Seaweed ExtractsCarrageenansSold under Carra™ trade nameProduced in Chile at our strategic partner Gelymar
Brown Seaweed ExtractsAlginatesSold under Satialgine™, AlgoGel™ and Cecalgum™ Produced at Algaia’s French factory in Lannilis
Tailor made solutionsCustomized SolutionsSold under Algogel™ tradename Produced in France
R&D services with partnersAlgaia ServicesBiomass, analytical, bioactivity etc…Provided from our R&D center in Saint Lô (France)
Specialty Seaweed extractsUnder Developmentwith specific partners or through JDATo be produced at Algaia’s French facility in Lannilis
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Production facility at LannilisNext to Brest
▪ 80,000 m2 land space – 15,000 m2 occupied▪ Over 40 years experience in Alginate production▪ ISO 9001 since 1994▪ FSSC 22000 certification
Algaia™ at a glance
R&D center at Saint LôNext to Caen
▪ 1000 m2 state-of-the-art R&D facility ✓ Extraction laboratory✓ Analytical laboratory✓ Application laboratory✓ Pilot room (in development)
Headquarter in Paris
Strategic Alliance with GelymarLocated in Chile
▪ 4th largest Carrageenan producer globally▪ ISO 9001 ▪ FSSC 22000 certification▪ Pharma cGMP production line
Saint Lô
Lannilis
Paris
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Laminaria digitata
Laminaria hyperborea
➢ France, 2nd processor country of seaweedin volumes in Europe (about 80,000 Ton/year)
➢ Algaia: 40,000 Tons of fresh seaweed/ year
Algaia™ at a glance
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Functional Systems
Carrageenansin Europe
InnovativeNatural
Ingredients
Red Seaweed& other natural
extracts
Algaia’s Services
Brown Seaweed
AlginatesgloballySpecialties
GreenBiorefineryAlgogel Customized™ Brand Name
Carra™ Brand Name
AnalyticalBiomass Potential
Bioactivity
R&D services
Process development
. Satialgine™ range
. Algogel™ range
. Cecalgum™ range
Upon Cargill’s acquisition :
Enhance functionalities
Algaia™ at a glance
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ALGAIA Services – Analyses & BioactivitiesAnalyses
Saccharides• Total sugars• Identification and quantification of polysaccharides• Characterisation in the composition• Study of the size range
Antioxidant compounds• Phenolic compounds• Vitamins• Pigments
Ionic Profile• Anions• Cations• Heavy metals• Isotopic ICPMS
Proteins• Total Proteins• Amino Acids
Pigment profile• Chlorophylls• Fucoxanthin• Astaxanthin• Phycocyanin• ….
Genomic identification• Microalgae strains• Seaweeds• ….
Fatty acid profile• Omega 6• Omega 3• Phospholipids• …
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ALGAIA Services – Analyses & BioactivitiesBioactivities
Prebiotic• Resistance to gastric enzymes • Selective fermentation of intestinal bacteria strains
Anti-microbial• Antiviral• Antibacterial• Antifungal
Anti-inflammatory• Enzymatic assays• Cytokines, interleukines• ….
Plant biostimulation• Germination• Defense elicitors• Plant nutrition / Stress• Plant defense gene expression
Anti-cholesterol• Influence on activity of selected genes involved in cholesterol absorption • Influence on activity of selected genes involved in cholesterol refining• Intestinal Absorption of cholestérol in vitro
Anti-cancer• Influence on selected genes involved in apoptosis• Proliferation• Adhesion• Migration and invasion• High flow screening
Cosmetic• Anti-ageing• Anti-UV• Antioxidant• Depigmentation• Cicatrisation
Cardiovascular• Atherosclerosis• Heart failure• hypertension• Obesity• Diabetes
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ALGAIA Services – boost R&D and markets
Culture/Wild stock of Macro & Micro algae
Producers of algal extractsUsers of algal extracts
Active formulae
• Analyses of batch composition in production
• Study on batch variability• Statistic studies on influence of
key cultivation parameters• Study on the valorisation of the
production• Support on potential markets of
interest• Support on wild biomass
assessment and management• Support in cultivation system
setup
• Studies on compatibility of extracts in formulated products
• Studies on products stability based on algal extracts
• Test on bioactivities of formulated products
• Identification of active algal extracts
• Development of novel algal extracts
• Analyses of batch composition in extract production
• Study on batch variability• Statistic studies on influence of
key production parameters• Tests on extract bioactivities• Valorisation of by-products
Get into sustainable commercial developmentas soon as possible !
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ALGAIA – First Biorefinery studies
Macrocystis pyrifera
Lessonia nigrescens Lessonia trabeculata
Alginates
Polyphenols
Oligo-elements
Laminarans
Fucoidans
Iodine
Dietary Fibers
Pigments
Proteins
Lipids
Biorefinery / Cracking
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Brown Seaweeds Main Active Ingredients known
Proportionsin Dry Matter
Plant Biostimulant(liquids)
Feed Applications Food / dieataryApplications
Personal Care
Sugars(carbohydrates)
(total > 60%)
Alginates / Uronic ac.
> 30% Binding capacity, contact effects, root (?)
Binders / Stabilisers
Binders / Stabilisers
Binders / StabilisersCicatrisation
Laminarans 1 - 20% Elicitors, source of glucose bacteria
Elicitor, Prebiotics Elicitor, Prebiotics
unknown
Fucoidans 1 - 20% Elicitors, bindingcapacity, Antioxidants
Anti-microbial,Anti-inflammatory
Anti-microbial Skin Protection
Fucoxanthins 1 - 4% Unknown Dietary fibers Dietary fibers Slimming
Mannitol 1 - 20% Osmoregulant, antioxidant
Antioxidant Antioxidant,Cholesterol
Antioxidant
Ashes(Sea Minerals)
5 - 30% Oligo-element support, root physiology (?)
Oligo-elements+ Iodine
Oligo-elements+ Iodine
Natural mineral salts
Proteins / AA 2 - 8% Low amounts in liquids Marine Protein / Peptides
Marine Protein/ Peptides
Unknown
Phlorotannins 2 - 15% Antioxidant Antioxidants Antioxidants Antioxidants
Lipids(fatty acids)
2 - 5% Low amounts in liquids Marine lipids Marine lipids Oily solutions
Biorefinery / Cracking
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Feed Health (incl. aqua)Preventing shrimps diseasesPreventing chicken diseasesAntibiotic alternative
Could be the most active component of seaweed extracts used as Biostimulants
Anti-inflammatoryPersonal Care
Ex: anti eczema
Pharma applications (multiple)
Anti-microbial
COLLOIDSFunctional Systems
Biorefinery / Cracking
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Cracking
Extraction
Raw material Seaweed
Liquid
Fucoidan Others
Solid
Alginate
Objective:
Co-extraction proof of concept
2 to 3 products of interest
1st yield & feasability evaluation
Products quality characterisation
Others
Alginate
Fucoidan
Material & Methods
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Experimental design
Soft aqueous extraction
(T°C, H2O, Time h)
Raw material
Liquid fraction(Fucoidan)
Alginic acid protocol
Solid fraction(Alginate)
Ca neutralisation
Na Alginate
Raw material
Alginic acid protocol
Ca neutralisation
Na Alginate
Single extraction Co-extraction
X3 X3
SeaweedSpecies
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Sampling overview
Protocole development + Evaluation Macrocystis
pyrifera
Single extraction Co-extraction
Frozen Dried Frozen Dried
15 samples(alginate)
9 samples(alginate)
cracking
6 samples(alginate)
27 samples(fucoïdans)
12 samples(alginate)
45 samples(fucoïdans)
Lessonianigrescens
Protocole development + Evaluation
Single extraction Co-extraction
Frozen Dried Frozen Dried
6 samples(alginate)
6 samples(alginate)
cracking
6 samples(alginate)
27 samples(fucoïdans)
Total Samples studied >220 samples + FRESH SAMPLES
12 samples(alginate)
36 samples(fucoïdans)
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Macrocystis pyrifera – Classical extraction versusCo-extraction
Products yieldsMacrocystis pyrifera (frozen)
Trial 1 Trial 2 Trial 3 Average
Alginate yield, single extraction (% dry weight)
13 13.2 11 12.4
Alginate yield, co-extraction (% dry weight)
25.3 14 26.9 22.1
Fucoidan yield, co-extraction(% dry weight)
1.89 1.71 2.27 1.96
➢ Better extraction yields of Alginates with co-extraction ?
➢ Fucoidan extraction yields quite low
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Macrocystis pyrifera – Classical extraction versusCo-extraction
➢ Positive effect on final product[color and yield]
whitening = current challenge in alginate industry
➢ Significative quality difference= low viscosity
Extract viscosityMacrocystis pyrifera (frozen)
Trial 1 Trial 2 Trial 3 Average
Single alginate extraction (cPo at 60rpm)
512 1330 400 747
Alginate co-extraction (cPo at 60rpm)
28 20 25 24
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Experimental data Control
Alginate co-extraction yield
(% MS)15.8
Fucoïdans co-extraction yield
(% MS)-
Alginate viscosity (cPo)
1% at 25°C169
6h/X°C
25.6
2.53
127
3h/2X°C
26.6
2.78
145
1h/3X°C
26.9
2.69
114
1h/5X°C
32.1
3.48
78
Average values based on triplicats
➢ Improvement of final alginate viscosity (low viscosity)
➢ Pre-extraction step at 2X°C seems interesting
➢ Experimental conditions must be improved again regarding alginate polymerisation (MV)
Macrocystis pyrifera –Co-extraction at various conditions
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Importance of process condition on yields
Extraction
Controllable parameters(water, stirring intensity…)
Uncontrollable parameters(seaweed wash, age of seaweed…)
Studied parameters(time, temperature, ratio)
ResponsesExtraction yieldsExtract compositionPolysaccharide composition
EXPERIMENTAL PLAN SETUPS
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Importance of process condition on yields
PROPOSITION OF A MATHEMATICAL MODEL
Full factorial design: 2n experiments (3 factors: 23 = 8 extractions)
Effect calculation of extraction parameters:
Extractions done in triplicate: calculation of confidence interval (t.σ𝑠2
3
𝑘)
Definition of influential parameters: effect ± confidence interval ≠ 0
𝑥𝑎 =(−𝑌1 − 𝑌2 − 𝑌3 − 𝑌4 + 𝑌5 + 𝑌6 + 𝑌7 + 𝑌8)
8
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Importance of process condition on yields
Ash content Phenolic compounds content Mannitol content
Fucoidan content Laminaran content
0.26±0.64
0.08±0.56
-0.81±0.67
0.66±0.27
-0.58±0.61
EXPERIMENTAL PLAN SETUPS – Effect of extraction time
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Importance of process condition on yields
EXPERIMENTAL PLAN SETUPS – Effect of extraction temperature
Fucoidan content Laminaran content
Ash content Phenolic compounds content Mannitol content
-0.09±0.64-0.36±0.56
1.88±0.67
1.46±0.27
-2.68±0.61
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Importance of process condition on yields
Fucoidan content in extract expressed as a percentage of the total solid content (n=3)
-1 -1
+1
+1
Time Temperature Ratio
3.84%
5.16%
4.50%4.50%
5.97%
3.04%
4.50%
+0.66% + 1.46% + 0.00%
Tim
e
Temperature
Ratio
0.35%
2.30%
1.51% 1.13%
2.81%3.05%
Time/temperature = +0.49%Time/ratio = -0.09%Temperature/ratio = -0.06%
% Fucoidan = 4.50% + 0.66%A + 1.46%B + 0.00%C + 0.49%AB – 0.09AC – 0.06%BC
Confidence interval at 0.95% calculated = 0.27%
% Fucoidan = 4.50% + 0.66%A + 1.46%B + 0.49%AB
EXPERIMENTAL PLAN SETUPS – Effect of combined parameters
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IN SUMMARY
➢Co-extraction of functional alginates and fucoidan is viable
➢Extraction conditions affect yields and chemical functionality
➢Temperature, H2O Ratio & Extraction time must be assessed
➢All these elements need to be studied before industrial & marketpositionning to avoid business downturns