LIFE 12 ENVIT352 Action C 5 Monitoring of

LIFE 12 ENV/IT/352 Action C 5 - Monitoring of the benefits from dyeing leather with naturalised colorants. Determination of trace elements in commercial dyes, crude chromophore, purified dyes and naturalized dyes Alessandro D’Ulivo BIONAD 24 Month Meeting January 27 th, 2016, UNIFI Premises , Florence (I)

LIFE 12 ENV/IT/352 Analized Dyes & Chromophore Crude Chromophore Commercial Dye YELLOW Disperse Yellow 42 ORANGE [Crude Chromophores + Additives ] Purified Chromophore Naturalized Dyes [From commercial Dyes] [from Purified Dyes] Teratop Brown GWL Disperse Yellow 42 DY 42 Nat Disperse Orange 30 Foron Brown Yellow S-2 RFL 150 Disperse Orange 30 D 030 Nat RED Disperse Red 202 Foron Brilliant Red S-RGL 200 Disperse Red 202 DR 202 Nat BLUE NA* Teratop Blue GLF Disperse Blue 27 DB 27 (*) Not commercially available BIONAD 24 Month Meeting January 27 th, 2016, UNIFI Premises , Florence (I)

LIFE 12 ENV/IT/352 Trace element Determination • Pressurized MW Digestion, HNO 3 + H 2 O 2 • Determination by Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES) Perkin Elmer Optima 8000 Elements: Hg, Cd, Pb, Tl Mn, Cr, V, Co, Ni, Cu, Zn B, As, Sb, Se Be, Ba Al, Fe BIONAD 24 Month Meeting January 27 th, 2016, UNIFI Premises , Florence (I)

LIFE 12 ENV/IT/352 Direct Determination Mercury Determination at of sub. Mercury p. p. b. Level Direct determination of Hg in solid and liquid samples Digestion not required No Hg losses No Hg contamination Typical Sample size 100 - 500 mg • L. O. Q < 20 pg Hg (< 0. 2 ppb) • DMA-80: fully compliant with US EPA method 7473 and with ASTM method D-6722 -01. • • • BIONAD 24 Month Meeting January 27 th, 2016, UNIFI Premises , Florence (I)

LIFE 12 ENV/IT/352 Results of Mercury Direct Determination Indicators of potential hazard level Detected Elements in Hg, Se, Pb, Cr, Cd, Pb, V, Be, at least one of the Dyes Ni, Mn, Zn, Al, Cu, Fe, B, Ba Elements Non-Detected in any dyes Tl 0. 17 ; Co 0. 17 (mg/kg) BIONAD 24 Month Meeting Above Limit of quantification L. O. Q. = 10 s Below the reported L. O. Q. January 27 th, 2016, UNIFI Premises , Florence (I)

LIFE 12 ENV/IT/352 Commercial Dyes Additives Commercial «Crude» Chromophores Soxhlet Extraction Purified Chromophores BIONAD 24 Month Meeting Lactose Naturalised Dyes January 27 th, 2016, UNIFI Premises , Florence (I)

LIFE 12 ENV/IT/352 Commercial Dyes Element Al As B Ba Be Cd Co Cr Cu Fe Hg Mn Ni Pb Sb Se Tl V Zn Totalb Teratop Brown GWL Crude Chromophores LOQm Foron Brilliant Red S-RGL 200 (mg/kg) Teratop Blue GLF Disperse Yellow 42 Disperse Orange 30 Disperse Red 202 (mg/kg) Foron Brown Yellow S-2 RFL 150 (mg/kg) (mg/kg) 32. 7 49. 8 47. 6 26. 1 64. 3 - - 0. 83 8. 1 -a 7. 5 - 0. 94 - - 0. 83 - - - 64. 1 3. 6 - 29. 0 0. 83 0. 84 0. 63 4. 8 3. 0 1. 7 - - 0. 02 - - - 0. 30 - 0. 07 0. 10 0. 13 - - 0. 15 - - 0. 08 - - - - 0. 17 - 3. 8 3. 3 2. 8 0. 59 0. 60 0. 17 - - - 11. 4 0. 40 1. 4 0. 33 21. 5 49. 9 44. 5 112. 4 29. 2 1. 8 33. 7 0. 08 0. 0070 0. 46 0. 0017 0. 66 0. 060 0. 010 0. 090 8. 5 10 -5 11. 2 18. 3 12. 9 59. 1 0. 91 0. 25 0. 34 0. 08 - 2. 4 2. 1 0. 42 - - 3. 5 2. 0 1. 5 1. 3 0. 83 2. 1 1. 9 - - - 1. 70 3. 9 2. 2 2. 0 - 2. 4 - 1. 70 - - - - 1. 70 9. 8 14. 3 22. 1 1. 8 0. 73 1. 1 2. 9 - - 10. 6 4. 6 0. 62 1. 6 0. 17 97. 7 148. 6 147. 1 299. 4 114. 8 13. 2 75. 1 [10. 4]c 0. 42 a - concentrations that are not reported are below LOQm. b - sum of concentrations includes LODm whenever the determined values cannot be quantified. c - sum of LOQm. BIONAD 24 Month Meeting January 27 th, 2016, UNIFI Premises , Florence (I)

LIFE 12 ENV/IT/352 Purified Chromophores from Commercial Dyes Element Al As B Ba Be Cd Co Cr Cu Fe Hg Mn Ni Pb Sb Se Tl V Zn Totalb Disperse Yellow 42 (mg/kg) 8. 2 0. 37 0. 13 0. 00015 3. 7 6. 8 25. 3 Disperse Orange 30 (mg/kg) 0. 0012 1. 7 10. 4 Naturalised Dyes Disperse Red 202 (mg/kg) 3. 9 0. 0001 12. 6 Disperse Blue 27 (mg/kg) 1. 1 1. 0 2. 0 0. 0030 0. 50 3. 7 17. 6 LOQm DY 42 Nat D 030 Nat DR 202 Nat DB 27 Nat (mg/kg) 0. 31 0. 85 1. 7 0. 010 2. 2 17. 7 31. 0 (mg/kg) 0. 90 0. 03 0. 29 0. 45 1. 0 0. 020 0. 19 1. 3 2. 8 13. 0 (mg/kg) 6. 6 0. 17 0. 27 4. 5 49. 7 0. 010 0. 57 2. 1 1. 2 6. 4 71. 4 (mg/kg) 0. 19 2. 1 0. 020 0. 25 1. 2 1. 0 15. 5 97. 7 a - concentrations that are not reported are below LOQm; b - sum of concentrations includes LODm whenever the determined values cannot be quantified; c - sum of LOQm BIONAD 24 Month Meeting January 27 th, 2016, UNIFI Premises , Florence (I) (mg/kg) 0. 83 0. 02 0. 07 0. 08 0. 17 0. 33 0. 08 8. 5 10 -5 0. 08 0. 42 0. 83 1. 70 0. 42 0. 17 [10. 4]c

LIFE 12 ENV/IT/352 Non quantifiable Elements (Conc ≤ LOQm) BIONAD 24 Month Meeting January 27 th, 2016, UNIFI Premises , Florence (I)

LIFE 12 ENV/IT/352 Direct. Total Determination of Mercury Element Content BIONAD 24 Month Meeting January 27 th, 2016, UNIFI Premises , Florence (I)

LIFE 12 ENV/IT/352 Direct Determination Mercury of Mercury BIONAD 24 Month Meeting January 27 th, 2016, UNIFI Premises , Florence (I)

LIFE 12 ENV/IT/352 Conclusion In general total elements content follow the order: Commercial Dyes > Crude Chromophore ≥ Naturalized Dyes > Purified Dyes Similar trend is found for mercury. Contamination of commercial dyes can be addressed to the use of additives with low degree of purity. Naturalized dyes, presents some problems of contamination by Zn, Fe and Al, probably arising from the apparatus employed in synthesis/purification. BIONAD 24 Month Meeting January 27 th, 2016, UNIFI Premises , Florence (I)