METHODOLOGY

The entire process of the laboratory experiment is discussed in this chapter

utilizing economical materials and agricultural waste specifically mango peel.

Material Selection

Selection of the materials was mostly based on categories such as

availability and workability. This laboratory experiment utilized the following

resources:

1. 15 pieces one-click bottles 12. Graduated Cylinders

2. Mango peel 13. Marking pen

3. Active dry yeast 14. Mortar and Pestle

4. Beakers 15. Paper

5. Bolo/knife 16. Pen

6. Bunsen Burner 17. pH paper

7. Chopping board 18. Scissors

8. Digital Weighing Scale 19. Scotch tape

9. Distilling Flask 20. Stirrer

10. Erlenmeyer Flask 21. Vegetable oil

11. Funnel 22. Water

 Procedural Framework

By referring to Figure 2, which shows the entire process undergone by the

biomass for bioethanol production, it is divided into four main sections:

Pretreatment, Ensiling, Fermentation and Distillation.

Biomass

Fresh mango peelings were gathered from waste products by fruit

vendors. Foreign materials were removed by washing the peelings and then air-

dried to 10% moisture content.

Pretreatment

Pretreatment section involves milling or size reduction of the mango

peelings by chopping the substrate into 3mm – 5mm sizes to aid the breakdown

of lignocellulosic materials present in the peelings. Initial pH level of the material

was determined.

Ensiling

Pretreated peelings were ensiled for a week in the one-click bottles, to

promote hydrolysis of cellulose in the lignocellulosic biomass to produce reducing

sugars. Anaerobic conditions were ensured by compacting the peelings to fill the

pore spaces.

Fermentation

After the ensiling process, the hydrolysate from the ensiled material was

extracted by squeezing using a cheesecloth. Bottles showing contaminations

were excluded in the process. Collected hydrolysate was boiled for about 15

minutes to kill the pathogens present. After which, hydrolysate was cooled then
pH level was determined and adjusted to pH 5. It is then equally distributed in

five 125 ml erlemenyer flasks. This is followed by the alcoholic fermentation with

the use of active dry yeasts (two percent by weight of the hydrolysate) that have

the function of converting the fermentable sugars obtained. Few drops of

vegetable oil were also added and tightly capped with rubber cork to prevent

escape of gases necessary for enzyme activity during fermentation.

Distillation

Next, distillation is done to recover ethanol from the fermentation products.

Hydrolysate was placed in a distilling flask and heated up to 80 degree Celcius.

Finally, recovered bioethanol was tested.

START
Figure 4. Process if the Biogas Digester Works
Figure 3. Digester Design for Biogas Production
 Working Principle

The anaerobic digestion will take place in the six-liter reactor. When the

biomass is already placed, the reactor must be sealed air-tight to prevent oxygen

from entering. When anaerobic digestion occur, micro-organisms breaks down

complex biomass components to smaller, less complex molecules. In the setup

rice straw and swine manure is used, the micro-organism present inside will now

start to work in breaking down the tissues of the rice straw and the manure. This

reaction produces biogas and digestate. Digestate is what is left of the biomass

after anaerobic digestion and can be used as an organic fertilizer.

The Biogas that was produced has two main components: methane (CH 4)

and Carbon Dioxide (CO2). These gases will be collected by passing a small tube

into a six-liter gas collector which is filled with water. The more biogas produced

in the reactor the more pressure is generated, this pressure will push the water

into a calibrated container which is the Water Displacer where the volume of the

gas produced is measured. In this setup the volume of gas produced is equal to

the volume of the water displaced. Figure 5 below shows the principle of how the

biogas digester design works.

The process will continue as long as anaerobic digestion still takes place

in the reactor. The biogas will be trapped in the gas collector until it is used.
Figure 5. Working Principle of Rice Straw and Swine Manure Biogas Production