Optimizing the biofuel produced from sugar cane Bagasse

1. Optimizing the biofuel produced from sugar cane Bagasse Group 1- 034 Li Lidao (2O4) Wu Licheng (2p3) Jeremy Lee(2o1) Walter Kong (2o4)

2. Agenda • Objectives • Rationale of investigation • Hypothesis • Variables • Literature review • Materials and apparatus • Methodology • Results and discussion • Limitations • Extension • Bibliography

3. Objectives To optimize pretreatment time and hydrolysis method for making biofuel from sugarcane bagasse Specifically: • Time of pre-treatment • 30 min, 60 min, 90 min, 120 min, 150 min • Type of hydrolysis • Acid (H2SO4 ) • Enzyme (Cellulase)

4. Hypothesis • Ethanol can be derived from sugarcane bagasse • Hydrolysis using cellulase will produce higher amount of ethanol than hydrolysis using sufluric acid • The longer the reaction time, the higher amount of ethanol will be achieved

5. Rationale • Biofuel is a greener and renewable alternative to fossil fuels, which is depleting at an alarming rate • Sugarcane has a high cellulose content and a large amount of sugarcane bagasse are generated after its sugar content was extracted • It is possible to generate biofuel from cellulose, but the pretreatment methods are costly and responsible for 20% of the total cost • Although enzymatic hydrolysis is expected to produce higher amount of ethanol, it is more expensive than acid hydrolysis

6. Literature Review • Pre-treatment though essential in optimising biofuel yield, it is responsible for 20% of the total cost. (Bin, Y. , 2007) • pre-treatment using alkali, ammonia, dilute acid and steam are capable of producing up to 90% of the theoretical sugar yields (Parveen, K., 2009) • Enzymatic hydrolysis is more expensive than acid hydrolysis, but unlike acid, it does not require high temperature and pressure. (MOHIT S. MISHRA., 2010)

7. Variables

8. Materials and apparatus

9. Reagent tests For quantitative analysis of Sugar and Ethanol Content

10. DNS reagent test • Reagent test procedures : • 2ml of DNS + 1ml of sample + 7ml of deionized water • Blank: 2ml of DNS + 8ml of deionized water • Heat it at a boiling water bath for 10min • Test in a spectrometer of 540nm of wavelength • A calibration curve was plot using various concentrations of glucose solution • pH level of sample will be adjusted to 7 using NaOH or HCl

11. Calibration Curve

12. Potassium dichromate (VI) test 0.05M of acidified Potassium Dichromate (VI) Plot a calibration curve using various concentrations of ethanol standard solutions Di Water:0.5 ml + Acidified potassium dichromate (VI) : 2.5 ml  Heat in boiling bath for 15 min Sample: 0.5 ml + Acidified potassium dichromate (VI): 2.5 ml  Heat in boiling bath for 15 min 575nm

13. Calibration Curve

14. Overview of Methodology Acid Hydrolysis Fermentation Pre-treatment Enzymatic Hydrolysis Distillation

15. Pre-treatment: Break the bonds between cellulose and hemicellulose and remove the lignin

16. pretreatment

17. Test the Sugar content of the sample using the DNS reagent test

18. Sugar Yield after pre-treatment Time of pre-treatment (min)

19. Discussion of Results • Trend: downward • The sugar yield comes from the partial hydrolysis of cellulose by acid. • Our results proved that glucose degradation of acid occurs when pre-treated for too long a duration. • Pre-treatment should not be carried out for more than an hour. Time of pre-treatment (min)

20. Overview of Methodology Acid Hydrolysis Fermentation Pre-treatment Enzymatic Hydrolysis Distillation

21. Hydrolysis: To break down cellulose into sugar

22. Acid Hydrolysis

23. Test the Sugar content of the sample using the DNS reagent test

24. Sugar Yield after acid hydrolysis

25. Discussion of Results • Likely to be a downward trend, due to glucose degradation by acid • Time of pre-treatment does not affect the sugar content after hydrolysis significantly A maximum of 20% of the sugarcane bagasse has been converted to sugar as a result of the treatments. Acid added during hydrolysis can also break down the lignin, through out the process of hydrolysis

26. Overview of Methodology Acid Hydrolysis Fermentation Pre-treatment Enzymatic Hydrolysis Distillation

27. Adjustment OF pH

28. Insert Picture of Neutralisation

29. Enzymatic Hydrolysis

30. Test the Sugar content of the sample using the DNS reagent test

32. Sugar Yield after enzymatic hydrolysis

33. Discussion of results • The longer the time of pre-treatment, the more lignin is removed and eases enzymatic hydrolysis • Enzymes (Cellulase) cannot remove the lignin preventing the break down of cellulose

34. Comparison of Enzymatic Hydrolysis and Acid Hydrolysis

35. Time of pre-treatment (min)

36. Discussion of results Acid Hydrolysis yields 610% more sugar than enzymatic hydrolysis

37. Overview of Methodology Acid Hydrolysis Fermentation Pre-treatment Enzymatic Hydrolysis Distillation

38. Adjustment OF pH

39. Fermentation- To convert sugar into ethanol

40. Overview of Methodology Acid Hydrolysis Fermentation Pre-treatment Enzymatic Hydrolysis Distillation

41. Distillation– To purify the ethanol

43. Conduct Potassium Dichromate (VI) Test for ethanol content

44. Ethanol Content – Acid and enzymatic Hydrolysis

45. Discussion of Results Acid Hydrolysis yields 205% more ethanol than enzymatic hydrolysis

46. Conclusions • Ethanol can be produced from sugarcane bagasse • The longer the time of pre-treatment, the greater the glucose yield when hydrolyzed by enzymes • The time of pre-treatment does not affect the glucose yield in the end when hydrolyzed by acid • Acid Hydrolysis is more effective than enzymatic hydrolysis at the given condition

47. Limitations • DNS reagent test cannot test non-reducing sugar which may be present after hydrolysis • DNS reagent test is sensitive to light and heat • Sugar cane bagasse in each sample slightly varies in its original sugar content

48. Extension • Produce biodegradable films from xylan extracts from sugarcane after pre-treatment. • Further investigate how does pre-treatment affect the xylan yield

49. Extraction of xylan