1. Naturalized Dyes Synthesis Bionad Life 12 Alicante (SPAIN), 07.07.2015 Serichim, 07/07/20151 Project LIFE12 ENV/IT/352 "Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle" (BIONAD)

2. Serichim, 07/07/20152 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 PREPARATIONSTATUS at 2/07/15 4kg SUGAR BLOCK2.3kg 2kg YELLOW NATYellow amide 2.3 kg 2Kg ORANGE NAT0 2Kg BLUE NAT0

3. Serichim, 07/07/20153 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)

4. Serichim, 07/07/20154 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

5. Serichim, 07/07/20155 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

6. Serichim, 07/07/20156 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 preparationgr Lactose.H2O4993 Isolated INT11943 Theorical INT1 (100% yield w/w) 6765 Expected INT1 (66% yield w/w) 4465 Yield% (w/w)29 1st preparationgr Lactose.H2O3996 Isolated INT13476 Theorical INT1 (100% yield w/w) 5411 Expected INT1 (66% yield w/w) 3571 Yield% (w/w)64 A third preparation is required

7. Serichim, 07/07/20157 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 preparationgr INT11983 INT2 41% ACN solution6266 INT22569 1st preparationgr INT13394 INT2 40% ACN solution10986 INT24394

8. 8Serichim, 07/07/2015 NMR 1H, INT2, batch 02/2015, pTsCl residue 1.1% w/w Second sugar block 30lt scale preparation

9. 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

10. 10Serichim, 07/07/2015 NMR 1H, INT2, batch 22/2015, pTsCl residue 0% w/w Second sugar block 30lt scale preparation

11. 11Serichim, 07/07/2015 PREPARATION OF INT3 Second sugar block 30lt scale preparation 1st (and 2nd°) preparationgr INT2 40% ACN Solution5440 INT3 (in 21.6% DCM solution)1726 INT2 Conversion (NMR INT2-INT3 molar ratio) 90 3rd preparationgr INT2 40% ACN Solution6266 INT3( in 16% DCM solution )1536 INT2 Conversion (NMR INT2-INT3 molar ratio) 70.5

12. 12Serichim, 07/07/2015 PREPARATION OF INT3 Second sugar block 30lt scale preparation NMR 1H, INT3, batch 04/2015,

13. 13Serichim, 07/07/2015 PREPARATION OF INT3 Second sugar block 30lt scale preparation NMR 1H, INT3, batch 23/2015,

14. 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,

15. 15Serichim, 07/07/2015 Yellow dye 30lt scale preparation Preparation of chromophore block Yellow 42 Coupling building blocks

16. 16 Preparation of chromophore block Yellow 42 Serichim, 29/01/2015

17. 17 Preparation of chromophore block Yellow 42 Serichim, 29/01/2015 5lt preparationgr Wet yellow 4217402 Yellow 42 ester isolated382 Yellow 42 ester Expected (100% yield w/w) 431 Yield % (w/w)88 HPLC purity %area98.7 30lt preparationgr Wet yellow 42171305 Yellow 42 ester 24% THF solution 1354 Yellow 42 ester Expected (100% yield w/w) 1398 Yield % (w/w)91 HPLC purity %area94

18. 18 Preparation of chromophore block Yellow 42 Serichim, 29/01/2015 5lt preparationgr yellow 42 ester382 Yellow 42 acid isolated321 Yellow 42 acid Expected (100% yield w/w) 357 Yield % (w/w)88 HPLC purity %area99 30lt preparationgr Yellow 42 ester (THF solution)1344 Yellow 42 acid988 Yellow 42 acid Expected (100% yield w/w) 1255 Yield % (w/w)78 HPLC purity %area88

19. 19 Preparation of chromophore block Yellow 42 Serichim, 29/01/2015

20. 20 Preparation of chromophore block Yellow 42 Serichim, 29/01/2015

21. 21 Preparation of chromophore block Yellow 42 Serichim, 29/01/2015 5lt preparationgr yellow 42 acid382 Residue with Yellow 42 amide 1130 Yellow 42 amide Expected (100% yield w/w) 738 Yield % (given)100 HPLC purity %area65.2 30lt preparationgr Yellow 42 acid988 Yellow 42 amide DCM solution17553 Yellow 42 amide Expected (100% yield w/w) 2280 Yield %100 HPLC purity %area63

22. Revised production plan ProductQuantity required, kg Done 7/07/15To be doneExpected time Sugar Synthon42.31.7September, 15 Yellow2(2.2)(0)July, 18 Orange22September, 15 Blue22October Project LIFE12 ENV/IT/352 "Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle" (BIONAD)