Horti-BlueC Lab scale extraction of chitin (Kaitlyn Vanhoutte & Johan Robbens, ILVO) Contents 1. Chitin 2. Chemical extraction 3. Chinese Mitten crab 4. Crangon crangon Chitin
• Major constituent of the exoskeleton (arthropods: e.g. insects, crustaceans and fungi)
• Polysaccharide • Long-chain polymer of N-acetylglucosamine Chemical Extraction
Crustacean shells • Shrimp (Crangon crangon) Nieuwpoort
• Chinese mitten crab Grobbendonk Chemical Extraction
Demineralization • Performed in dilute hydrochloric acid (HCl) solution. • Eliminate the calcium carbonate + calcium chloride. • Filtrated washed dried.
Demineralization Deproteinization acid treatment alkali treatment Chemical Extraction
Deproteinization • Performed in dilute sodium hydroxide (NaOH) • Alkaline treatment • Remove proteins. • Filtrated washed dried.
Demineralization Deproteinization acid treatment alkali treatment Chemical Extraction
Optional treatments • Deacetylation: Convert chitin to chitosan by removal of acetyl group.
• Decolorization process: Astaxanthin and β-carotene elimination. Chemical Extraction
Li, V. (2006, June 22). Green Saviour–Chitin and Chitosan [Powerpoint]. Retrieved from https://www.scribd.com/document/49729324/Green-Saviour-Chitin-and- Chitosan Chinese Mitten crab
Chemical Washing Extraction Eriocheir sinensis Shredding Chinese Mitten crab
Method 5 Method 6 Method 7
Start dry weight (g) 150,47 123,09 150,46 End dry weight (g) 14,06 14,12 12,83 Recovery (%) 9,34 11,47 8,53 Best treatment: Method 7 Deproteinization: 10% NaOH (25mL/g sample) Demineralization: 6% HCl (25mL/g sample)
• Untreated Chinese Mitten Crab contains a lot of N • Method 7 has the lowest phytotoxicity. Chinese Mitten crab
PAH analysis: results expressed as µg/kg
acenapthene fluorene phenanthrene 2,340 anthracene 0,303 fluoranthene 2,711 pyrene 0,817 benzo(a)anthracene 0,566 chrysene 0,453
benzo(b)fluoranthene 1,230
benzo(k)fluoranthene 4,176 benzo(j)fluoranthene 0,904 benzo(e)pyrene 0,338 benzo(a)pyrene 0,360
indeno(123cd)pyrene 0,193 benzo(ghi)perylene 0,374 Chinese Mitten crab
Dry matter:
Code m(recipiënt) (g) m(sample) (g) m(dry) (g) Dry matter (%)
CWK 101,79 18,98 107,68 31,05
Heavy metals:
Fe Zn Cu Cr Cd Pb Hg As Samples Horti-BlueC mg kg -1 mg kg -1 mg kg -1 µg kg -1 µg kg -1 µg kg -1 µg kg -1 µg kg -1 CWK 316,0 23,8 62,7 390,4 78,3 91,3 13,9 938 Chinese Mitten crab
PCB: PCB - ng/g fat CWK PCB - ng/g wet weight CWK HCB 37,2 HCB 0,55 CB28 10,2 CB28 0,15 CB31 32,1 CB31 0,48 CB52 50,2 CB52 0,75 CB101 239,5 CB101 3,57 pp'-DDE 352,6 pp'-DDE 5,25 CB118 271,1 CB118 4,04 CB153 904,8 CB153 13,48 CB105 68,1 CB105 1,01 CB138 551,8 CB138 8,22 CB156 66,5 CB156 0,99 CB180 367,8 CB180 5,48
Fat content (%) 1,49 Crangon crangon
Based on both yield as price differences, we choose method 4. Price Less chemical reagents: higher sustainability Average yield. Method 4 Deproteinization: 1M NaOH (1:10 w/v) 80 °C; 2 hours; 500rpm Demineralization: 1,5M HCl (1:10 w/v) room temperature; 3 hours; 500rpm HBC-W1-P-22_shriM4
Note: This choice of method doesn’t include the purity or characterization of the different types of chitin. Crangon crangon
Method 1 Method 2 Method 3 Method 4 Start wet weight 769,25 817,26 751,02 50,68 (g) Start dry weight 208,81 221,84 203,86 13,51 (g) End dry weight (g) 21,81 23,71 21,06 1,71 Recovery (%)* 10,44 10,69 10,33 12,66 HBC-W1-P-22_shriM4 HBC-W1-P-22_shriM4 HBC-W1-P-22_shriM4 HBC-W1-P-22_shriM4
* Recovery based on dry weight of the shrimp shells (C. crangon). Crangon crangon PAH analysis: results expressed as µg/kg GAR Chitin method 4 acenapthene acenapthene 0,511 fluorene fluorene 0,814 phenanthrene 1,702 phenanthrene 4,698 anthracene 0,209 anthracene 0,099 fluoranthene 1,926 fluoranthene 2,623 pyrene 0,383 pyrene <0.697 benzo(a)anthracene 0,372 benzo(a)anthracene 0,498 chrysene 0,389 chrysene 0,418 0,679 benzo(b)fluoranthene benzo(b)fluoranthene 0,440 0,428 benzo(k)fluoranthene benzo(k)fluoranthene 0,272 benzo(j)fluoranthene 0,343 benzo(j)fluoranthene 0,403 benzo(e)pyrene 0,399 benzo(e)pyrene 0,247 benzo(a)pyrene 0,255 benzo(a)pyrene 0,441 0,199 indeno(123cd)pyrene indeno(123cd)pyrene 0,139 benzo(ghi)perylene 0,398 benzo(ghi)perylene 0,304 Crangon crangon
Dry matter:
Code m(recipiënt) (g) m(sample) (g) m(dry) (g) Dry matter (%)
Crangon crangon 98,44 26,66 105,56 26,75
Heavy metals:
Ni Zn Cu Cr Cd Pb Samples Horti-BlueC mg kg -1 mg kg -1 mg kg -1 mg kg -1 mg kg -1 mg kg -1 Crangon crangon 12,4 86 53,6 73,5 0,09 1,1 Chitin method 4 1,1 16 4 1,5 0,02 0,6 Crangon crangon
PCB: PCB - ng/g fat Crangon crangon PCB - ng/g wet weight Crangon crangon HCB <4.0 HCB <0.07 CB28 <4.0 CB28 <0.07 CB31 <4.0 CB31 <0.07 CB52 <4.0 CB52 <0.07 CB101 <10 CB101 <0.18 pp'-DDE <4.0 pp'-DDE <0.07 CB118 <10 CB118 <0.18 CB153 <15 CB153 <0.26 CB105 <4 CB105 <0.07 CB138 <12 CB138 <0.21 CB156 14,6 CB156 0,26 CB180 8,8 CB180 0,15
Fat content (%) 1,76 Info? [email protected]
This project has received funding from the Interreg 2 Seas Programme 2014-2020 co-funded by the European Regional Development fund under subsidy contract No 2S03-046 Horti-BlueC