PROJECT: LIFE12 ENV/IT/352 «BIONAD»

<<Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle>>

DAVIDE PELLEGRINI ICCOM-CNR,

Pisa6 Months Progress Meeting – BIOKIMICA premises, Santa Croce sull’Arno (PI),

June 13th 2014

Actions 

B.1 BIOKIMICA, INESCOP, ICCOM-CNR(0-6 months)

Demonstration of the use of traditional chemical acid dyes in leather dyeing

B.2 BIOKIMICA, INESCOP, (ICCOM-CNR)(0-6 months)

Demonstration of the quality features of dyed leather with acid dyes

B.4 BIOKIMICA, (ICCOM-CNR)(3-9 months)

Demonstration of naturalised dyes in leather dyeing at laboratory level

DO30

DR202DY42

DB27

Naturalised dyes 

LACTOSE

Commercial acid dyes 

NN

NH2

OCH3

CH3

SO3Na

N

N

CH3

O

N

NH

SO3Na

NaO3S

NH

N N

N

O

CH3

O37 Y49

R249B113

Leather dyeing procedure

0.2 g 4 mg/2 mL2% (dye/leather)

Magnetic stirring at 20°C for 1 h

Heating at 55°C

400 µL MilliQ + 4 mg formic acid

Magnetic stirring for 0.5 h

Washing with 4 mL MilliQfor 3 times

1. Formic acid at 55°C

B.1, B.4

Leather dyeing procedure

Magnetic stirring at 20°C for 1 h

Neutralization with 50µL K2CO3 (pH = 7-8)

Magnetic stirring for 10 min

2. Neutralization

Washing with 4 mL MilliQfor 3 times

0.2 g

B.1, B.4

4 mg/2 mL2% (dye/leather)

Heating at 55°C

400 µL MilliQ + 4 mg formic acid

Magnetic stirring for 0.5 h

Dyes characterizationUV-Vis

Naturalised dyes

Commercial acid dyes

1 Dyeing bath2 Washing 13 Washing 24 Washing 3

B.1, B.2, B.4

FORMIC ACIDPROCEDURE

Naturalised dyes

Commercial acid dyes

1 Dyeing bath2 Washing 13 Washing 24 Washing 3

B.1, B.2, B.4

Dyes characterizationUV-VisNEUTRALIZATIO

NPROCEDURE

B.1, B.2, B.4

Dyes characterizationUV-Vis

200 250 300 350 400 450 500 550 600 650 700 750 800

-0.1

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

1.1

1.2

1.3

Wavelength [nm]

Abs

orb

anc

e

DO30 dyeing bathDO30 washing 1DO30 washing 2DO30 washing 3

0.0

0.5

Washing water 3

Washing water 2

Washing water 1

A

Tanning solution

-61%-79%

-83%

442

Dyed leather characterization

Naturalised dyes

Commercial acid dyes

2%

5%

10%

15%

B.1, B.2, B.4

Naturalised dyes

Commercial acid dyes

Formic acid

Neutralization

B.1, B.2, B.4

Dyed leather characterization

Dyed leather characterization

Colorimetry

B.1, B.2, B.4

CM2600d KONICA MINOLTA

360-740 nmα = 10°

D65d. 5 mm

SAMPLESCI SCE

L a b L a bDR202 acid 48.11 39.65 13.66 48.08 39.59 13.64DY42 acid 81.47 -7.6 25.79 81.35 -7.56 25.75DB27 acid 53.56 -13.29 -22.35 53.4 -13.29 -22.39DO30 acid 62.92 25.73 39.27 62.9 25.65 39.22Red acid 52.46 48.56 11.48 52.33 48.62 11.56

Yellow acid 78.02 -10.69 57.24 77.85 -10.62 57.18Blue acid 32.12 3.76 -22.11 31.96 3.79 22.21

Orange acid 55.68 32.81 40.07 55.56 32.87 40.19DR202 neutr 45.11 40.08 14.81 45.04 40.06 14.81DY42 neutr 80.83 -7.45 30.18 80.72 -7.41 30.14DB27 neutr 51.54 -13.61 -21.97 51.39 -13.61 -22.02DO30 neutr 60.93 27.44 40.41 60.92 27.37 40.35Red neutr 54.3 48.46 -11.38 54.19 48.52 -11.45

Yellow neutr 80.71 -11.06 47.98 80.6 -10.94 47.58Blue neutr 35.82 3.48 -23.8 35.73 3.54 -23.84

Orange neutr 64.6 2.14 38.04 64.5 24.12 38.03

FT-IR CHARACTERIZATION

05001000150020002500300035000

0.02

0.04

0.06

0.08

0.1

0.12

Wavenumbers [1/cm]

Abs

orb

anc

e

AMIDE IC=O ν

~1650 cm-1

AMIDE IICN ν, NH δ

~1550 cm-1

ATR spectrum128 scans

Untreated leather

COOH ν~1730 cm-1

B.1, B.2, B.4

FT-IR CHARACTERIZATION

05001000150020002500300035000

0.02

0.04

0.06

0.08

0.1

0.12

Wavenumbers [1/cm]

Abs

orb

anc

e

AMIDE IICN ν, NH δ

~1550 cm-1

ATR spectrum128 scans

Untreated leather

COOH ν~1730 cm-1

AMIDE I1~1654 cm-1

AMIDE I2~1636 cm-1

AMIDE IC=O ν

~1650 cm-1

B.1, B.2, B.4

162016251630163516401645165016551660

Wavenumbers [1/cm]

0.954

0.96

0.966

0.972

0.978

0.984

0.99

0.996

1.002

1.008

1.014

1.02

Abs

orba

nce

BLANKDO30-HCl

DY42-HClDR202-HClDB27-HCl

1654 cm-1 1650 cm-1 1634 cm-1 1637 cm-1

Naturalised dyes

FT-IR CHARACTERIZATION

B.1, B.2, B.4

162016251630163516401645165016551660166516700.965

0.97

0.975

0.98

0.985

0.99

0.995

1

1.005

1.01

Wavenumbers [1/cm]

Abs

orba

nce

BLANKORANGE

37YELLOW 49RED 249

BLUE 113

FT-IR CHARACTERIZATIONCommercial dyes

B.1, B.2, B.4

FT-IR CHARACTERIZATION

Principal Component Analysis

05001000150020002500300035000

0.02

0.04

0.06

0.08

0.1

0.12

Wavenumbers [1/cm]

Abs

orb

anc

e

AMIDE I1~1654 cm-1

AMIDE I2~1636 cm-1

VARIABLES

Peak heights of

AI1/AIIAI2/AII

COOH/AII

B.1, B.2, B.4

AMIDE IICN ν, NH δ

~1550 cm-1

COOH ν~1730 cm-1

AMIDE IC=O ν

~1650 cm-1

FT-IR CHARACTERIZATION

Principal Component Analysis

-2 0 2 4

-1.5

0.0

1.5

DO30-HCl DO30-HCl

Yellow

Orange

Red

Blue

Yellow

Orange

Red

Blue

DY42-HCl

DR202-HCl

DB27-HCl

DY42-HCl

DR202-HCl

DB27-HCl

PC1 (69%)

PC

2 (

29

%)

Leather

Blank

Blank

-1 0 1

-1

0

1

AI1/AII

AI2/AII

PC1

PC

2

COOH/AII

PCA

Variables: AI1/AII, AI2/AII, COOH/AII

Var. tot. = 98%Matrix: covariance

Score plot

Loading plotFormic acid procedure

Neutralization procedure

B.1, B.2, B.4

FT-IR CHARACTERIZATION

Principal Component Analysis

-2 0 2 4

-1.5

0.0

1.5

DO30-HCl DO30-HCl

Yellow

Orange

Red

Blue

Yellow

Orange

Red

Blue

DY42-HCl

DR202-HCl

DB27-HCl

DY42-HCl

DR202-HCl

DB27-HCl

PC1 (69%)

PC

2 (

29

%)

Leather

Blank

Blank

-1 0 1

-1

0

1

AI1/AII

AI2/AII

PC1

PC

2

COOH/AII

PCA

Variables: AI1/AII, AI2/AII, COOH/AII

Var. tot. = 98%Matrix: covariance

Score plot

Loading plot1 2

Formic acid procedure

Neutralization procedure

B.1, B.2, B.4

FT-IR CHARACTERIZATION

Principal Component Analysis

-2 0 2 4

-1.5

0.0

1.5

DO30-HCl DO30-HCl

Yellow

Orange

Red

Yellow

Orange

Blue

DY42-HCl

DR202-HCl

DY42-HCl

DR202-HCl

DB27-HCl

PC1 (69%)

PC

2 (

29

%)

Leather

Blank

Blank

-1 0 1

-1

0

1

AI1/AII

AI2/AII

PC1

PC

2

COOH/AII

PCA

Variables: AI1/AII, AI2/AII, COOH/AII

Var. tot. = 98%Matrix: covariance

Score plot

Loading plot1 2

1 High AI1/AII, AI2/AII

Formic Acid procedure(and RED

neutralization)2 Low AI1/AII, AI2/AII

Neutralization procedure

(and DB27-HCl and blue formic acid

procedure)

Formic acid procedure

Neutralization procedure

B.1, B.2, B.4

Blue

Red

DB27-HCl

FT-IR CHARACTERIZATION

Principal Component Analysis

1000110012001300140015001600170018001900

-0.1

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

1.1

Wavenumbers [1/cm]

Abs

orba

nce

DY42-HCl Formic

acidDY42-HCl

Neutralization

AMIDE II INCREASES

B.1, B.2, B.4

COLLAGEN CROSS-LINKING

FT-IR

~1654 cm-1/~1690 cm-1

Collagen cross-linking type

Pyridinoline (Pyr)

Dehydro-dihydroxy-norleucine (deH-

DHLNL)

B.1, B.2, B.4

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.516

54cm

-1/1

690c

m-1

Collagen cross-linking

Blank

Leather

Commercial

Naturalised

Naturalised dyes

Rich in Pyr type cross-

linking

COLLAGEN CROSS-LINKING

FT-IR

B.1, B.2, B.4

Naturalised dyes 5%, 10%,

15%

Same behavior of commercial dyes

2% 0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

1654

cm-1

/169

0cm

-1

Collagen cross-linking

COLLAGEN CROSS-LINKING

FT-IR

B.1, B.2, B.4

COLLAGEN CROSS-LINKING

TGA

100 200 300 400 500 60010

20

30

40

50

60

70

80

90

100

T (°C)

TG

%0

50

100

150

200

250

300

350

DT

G (

ug/m

in)

Gas: N2 200 mL/minHeating: 30-700°C, 10 °C/min

B.1, B.2, B.4

COLLAGEN CROSS-LINKING

TGA

100 200 300 400 500 60010

20

30

40

50

60

70

80

90

100

T (°C)

TG

%

0

50

100

150

200

250

300

350

DT

G (

ug/m

in)

T01

T02

Tonset

% fin

SAMPLE

T01 Tonset T0

2 % fin

Leather 77.4 302.2 334.3 22.2892BLANK 72.7 299.9 338 20.2782

DB27 2% 70.6 300.6 339.5 20.9291DO30 2% 76.6 298.2 337 21.5649DY42 2% 74 297.7 337.6 20.2527DR202

2%74.1 299.1 339 21.639

Blue 76.1 299.2 335.4 22.3998Orange 75.4 302 335.2 22.1174Yellow 71.9 300.6 333.5 20.4022Red 73 299.1 336 21.37

DB27 15%

72.7 299.3 330.9 22.6931

DO30 15%

77.7 297.4 334.6 22.4728

DR202 15%

79 299.9 330.6 22.1285

DY42 15%

76.5 299.5 331.5 22.5689

B.1, B.2, B.4

COLLAGEN CROSS-LINKING

TGA

B.1, B.2, B.4

326.0

328.0

330.0

332.0

334.0

336.0

338.0

340.0

342.0

.T 02

100 200 300 400 500 60010

20

30

40

50

60

70

80

90

100

T (°C)

TG

%

0

50

100

150

200

250

300

350

DT

G (

ug/m

in)

T01

T02

Tonset

% fin

Naturalised2%

Commercial2%

Naturalised

15%

FUTURE ACTIONS

B.1 BIOKIMICA, INESCOP, ICCOM-CNR(0-6 months)

Demonstration of the use of traditional chemical acid dyes in leather dyeing

HPLC-DAD characterization of dyes bath and washing solutions

GC-MS characterization of the volatile fraction of the dyed leather

C.1 ICCOM-CNR, UNIFI, INESCOP, BIOKIMICA(0-12 months)

Monitoring on the enviromental impact of acid dyes within leather manufacture

Atomic spectroscopy determination of the metal content of dyeing wastewaters (Cr, Co, Cu, Fe)

BIONAD PEOPLE @ ICCOM-CNR, PISA

Emilia BramantiAlessandro D’Ulivo

Massimo Onor

Manuela Cempini

Davide Pellegrini

Emanuela Pitzalis