REMOVAL OF BTX CONTENT IN DIESEL FUEL USING PSEUDOMONAS PUTIDA

Material
250 mL Erlenmeyer flasks
petri dish
inoculating needle or loop
graduated cylinder
beaker
burner
Ostwald Viscometer
Hydrometer
Pycnometer
Litmus paper
pH meter

Procedures:
Diesel Fuel
1. Twenty – five (25) mL diesel fuel will be poured into five 250 mL erlenmeyer flasks.
The erlenmeyer flasks with diesel fuel will be weighed and the data will be recorded
to determine the initial weight of the diesel fuel.
I. PHYSICAL PROPERTIES
A. Color
Observe the color of the liquid diesel fuel.
B. Density
Calculate the density of the liquid fuel.
i. Using mass-volume method
1. Using the analytical balance, measure the mass of the empty
beaker.
2. Measure a volume of 250 mL of liquid diesel fuel.
3. Measure the mass of beaker with diesel.
4. Calculate the mass of the diesel.
5. Calculate the density of the diesel.
ii. Using hydrometer
1. Measure a volume of 250 mL of diesel fuel.
2. Put the hydrometer in the liquid.
3. Measure the readings.
4. Compare the measured density to the standard.
C. Odor
Observe the odor of the liquid diesel fuel.
D. pH
i. Using Lithmus Paper
1. Have three trials in measuring the pH meter using lithmus paper.
2. Compare the result into the standard pH.
ii. Using pH meter
1. Have three trials in measuring the pH of the liquid diesel fuel using
pH meter.
2. Compare the result into the standard pH.
E. Viscosity
 i. Using viscometer
1. Set up the temperature into ambient air measurement.
2. Measure 250 mL of water to the lower reservoir of viscometer and
put in the thermostat.
3. Measure the mass of empty pycnometer.
4. Fill the pycnometer with 500 mL of water and put in the
thermostat.
5. If the temperature is stable, measure the time of outflow.
6. Remove the water in the viscometer and fill it with liquid diesel.
7. Measure the mass of pycnometer with water.
8. Clean the pycnometer and fill it with liquid diesel fuel.
9. Measure the outflow.
10. Measure the mass of pycnometer with diesel.
11. Calculate the density of diesel fuel using the formula:
𝑚��+𝑠 − 𝑚𝑝
��𝑠 = ��
𝑚𝑝+𝑤 − 𝑚𝑝 𝑤

II. CHEMICAL PROPERTIES

The presence and the amount of volatile organic compounds and saturated
hydrocarbons in the treated diesel fuel will be analyzed using High Performance
Liquid Chromatography (HPLC). The sample would be tested in DOE.

Preparation of New Culture of Pseudomonas putida f1 strain

1. Working cultures will be prepared using nutrient agar, petri dish, and inoculating
needle or loop. To transfer the bacteria to the petri dish, inoculating needle or loop
will be used.
2. The nutrient agar will be subjected to the bottom side of the petri dish.
3. Sterilization will be done by having an auto sterilized petri dish and by contacting
the inoculating needle in flame from the burner during transferring of the bacteria.