Naturalized Dyes Synthesis Bionad Life 12

Alicante (SPAIN), 07.07.2015

Serichim, 07/07/2015 1

Project LIFE12 ENV/IT/352"Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle" (BIONAD)

Serichim, 07/07/2015 2

Project LIFE12 ENV/IT/352"Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle" (BIONAD)

Sequence of 2° naturalized dye campaigne

PREPARATION STATUS at 2/07/15

4kg SUGAR BLOCK 2.3kg

2kg YELLOW NAT Yellow amide 2.3 kg

2Kg ORANGE NAT 0

2Kg BLUE NAT 0

Serichim, 07/07/2015 3

Project LIFE12 ENV/IT/352"Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle" (BIONAD)

SERICHIM ACTIVITIES FROM MAY 13th TO JULY 3rd

• Second sugar block 30lt scale preparation• Second yellow amide preparation (30lt scale)

Serichim, 07/07/2015 4

Project LIFE12 ENV/IT/352"Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle" (BIONAD)

Second sugar block 30lt scale preparation

• 30lt jacketed stirred tank reactor equipped

with condenser, vacuum pump,

thermocouple, vacuum controller, heating

and cooling device

Serichim, 07/07/2015 5

Project LIFE12 ENV/IT/352"Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle" (BIONAD)

Second sugar block 30lt scale preparation

O

O

O OH

O

OH O O

O

O

CH3

CH3

CH3

CH3CH3

CH3

OMeMeO

O

OH

OH OH

O

OHO

OH

OH

OH

OH

H2OINT 1

O

O

O OH

O

TsO O O

O

O

CH3

CH3

CH3

CH3CH3

CH3

OMeMeO

INT 2

O

O

O OH

O

N O O

O

O

CH3

CH3

CH3

CH3CH3

CH3

OMeMeO

NH

INT 3

1° STEP

2° STEP

3° STEP

Serichim, 07/07/2015 6

Project LIFE12 ENV/IT/352"Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle" (BIONAD)

Second sugar block 30lt scale preparation Preparation of INT1

2nd preparation gr

Lactose .H2O 4993

Isolated INT1 1943

Theorical INT1 (100% yield w/w)

6765

Expected INT1 (66% yield w/w)

4465

Yield% (w/w) 29

1st preparation gr

Lactose .H2O 3996

Isolated INT1 3476

Theorical INT1 (100% yield w/w)

5411

Expected INT1 (66% yield w/w)

3571

Yield% (w/w) 64

A third preparation is required

Serichim, 07/07/2015 7

Project LIFE12 ENV/IT/352"Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle" (BIONAD)

Second sugar block 30lt scale preparation Preparation of INT2

2nd preparation gr

INT1 1983

INT2 41% ACN solution 6266

INT2 2569

1st preparation gr

INT1 3394

INT2 40% ACN solution 10986

INT2 4394

8Serichim, 07/07/2015

NMR 1H, INT2, batch 02/2015, pTsCl residue 1.1% w/w

Second sugar block 30lt scale preparation

9Serichim, 07/07/2015

NMR 1H, INT2, batch 02/2015, pTsCl residue 1.1% w/w

Second sugar block 30lt scale preparation

NMR 1H, INT2, batch 22/2015, pTsCl residue 0% w/w

10Serichim, 07/07/2015

NMR 1H, INT2, batch 22/2015, pTsCl residue 0% w/w

Second sugar block 30lt scale preparation

11Serichim, 07/07/2015

PREPARATION OF INT3

Second sugar block 30lt scale preparation

1st (and 2nd°) preparation gr

INT2 40% ACN Solution 5440

INT3 (in 21.6% DCM solution) 1726

INT2 Conversion (NMR INT2-INT3 molar ratio)

90

3rd preparation gr

INT2 40% ACN Solution 6266

INT3( in 16% DCM solution ) 1536

INT2 Conversion (NMR INT2-INT3 molar ratio)

70.5

12Serichim, 07/07/2015

PREPARATION OF INT3

Second sugar block 30lt scale preparation

NMR 1H, INT3, batch 04/2015,

13Serichim, 07/07/2015

PREPARATION OF INT3

Second sugar block 30lt scale preparation

NMR 1H, INT3, batch 23/2015,

14Serichim, 07/07/2015

PREPARATION OF INT3

Second sugar block 30lt scale preparation

NMR 1H, INT3, batch 23/2015,

NMR 1H, INT3, batch 04/2015,

15Serichim, 07/07/2015

Yellow dye 30lt scale preparation

• Preparation of chromophore block Yellow 42

• Coupling building blocks

16

• Preparation of chromophore block Yellow 42

NH

SO O

NO2

N

CO2H

Yellow 42 acid

NH

SO O

NO2

N

CO2Et

Yellow 42 ester

NH

SO O

NO2

NH

Yellow 42

STEP 1

STEP 2

Serichim, 29/01/2015

17

• Preparation of chromophore block Yellow 42

Serichim, 29/01/2015

5lt preparation gr

Wet yellow 4217 402

Yellow 42 ester isolated 382

Yellow 42 ester Expected (100% yield w/w)

431

Yield % (w/w) 88

HPLC purity %area 98.7

30lt preparation gr

Wet yellow 4217 1305

Yellow 42 ester 24% THF solution

1354

Yellow 42 ester Expected (100% yield w/w)

1398

Yield % (w/w) 91

HPLC purity %area 94

18

• Preparation of chromophore block Yellow 42

Serichim, 29/01/2015

5lt preparation gr

yellow 42 ester 382

Yellow 42 acid isolated 321

Yellow 42 acidExpected (100% yield w/w)

357

Yield % (w/w) 88

HPLC purity %area 99

30lt preparation gr

Yellow 42 ester (THF solution) 1344

Yellow 42 acid 988

Yellow 42 acid Expected (100% yield w/w)

1255

Yield % (w/w) 78

HPLC purity %area 88

19

• Preparation of chromophore block Yellow 42

Serichim, 29/01/2015

Yellow 42 acido

+

NNH

O

O

O OH

O

O O

O

O

CH3

CH3

CH3

CH3CH3

CH3

OMeMeO

Latt-Prot Pip

DCM, under N2, ov

WSC HCl

Yellow 42 -Lattosio protetto

NH

SO O

NO2

N

CO2H

NH

SO O

NO2

N

CO

N

N

O

OO

OH

O O

OO

O

CH3CH3

CH3

CH3

CH3CH3

MeO

OMe

20

• Preparation of chromophore block Yellow 42

Serichim, 29/01/2015

NH

SO O

NO2

N

CO

N

N

O

OO

OH

O O

OO

O

CH3CH3

CH3

CH3

CH3CH3

MeO

OMe

Yellow 42 -Lattosio protetto

NH

SO O

NO2

N O

N

N

O

OH

OH

OH

O

O

OH

OH

OH

OH

HCl 37% in acetone

acetone, 0-ta, ov

+ 2MeOH

+ 3MeCOMe

YELLOW

21

• Preparation of chromophore block Yellow 42

Serichim, 29/01/2015

5lt preparation gr

yellow 42 acid 382

Residue with Yellow 42 amide

1130

Yellow 42 amideExpected (100% yield w/w)

738

Yield % (given) 100

HPLC purity %area 65.2

30lt preparation gr

Yellow 42 acid 988

Yellow 42 amide DCM solution 17553

Yellow 42 amideExpected (100% yield w/w)

2280

Yield % 100

HPLC purity %area 63

Revised production plan

Product Quantity required, kg

Done 7/07/15 To be done Expected time

Sugar Synthon 4 2.3 1.7 September, 15

Yellow 2 (2.2) (0) July, 18

Orange 2 2 September, 15

Blue 2 2 October

Project LIFE12 ENV/IT/352"Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle" (BIONAD)