Veronica Chang

Chem 1A Lab
16 November 2015
Erin Sullivan
Biofuels Unit: Eco-toxicity Assay, Biodiesel Synthesis and Combustion
Abstract:
In order to determine the feasibility of biodiesel as an alternative energy source, the
efficiency and toxicity of biodiesel was compared to those of other biofuels and diesel. Biodiesel
was synthesized and combusted, and its heat of combustion was measured using calorimetry.
Viscosity was determined by recording the time it took for a fuel to pass through a Pasteur
pipette. The eco-toxicity assay was prepared by adding biofuel solutions of different
concentrations to petri dishes with radish seeds and observing the percent of seeds that
germinated. The data showed that lab synthesized biodiesel and commercial biodiesel were less
toxic than other biofuels; there was 0% germination of seeds at 10% concentration by weight
volume of methanol, ethanol, and 2-butanol, while at the same concentration 95% germinated in
lab biodiesel solution and 54.5% germinated in commercial biodiesel. In addition biodiesel was
found to be more viscous than methanol and diesel; lab synthesized biodiesel and commercial
biodiesel took 8.90 seconds and 5.5 seconds, respectively, to pass through a Pasteur pipette,
whereas methanol took 1.00 seconds and diesel took 2.96 seconds. Also, commercial biodiesel
and lab synthesized biodiesel had lower heats of combustion, -34.6 kJ/g and -29.647 kJ/g
respectively, than diesel which had a heat of combustion of -46.7 kJ/g.

Introduction
The purpose of this lab was to determine the efficiency and toxicity of biodiesel in
comparison with other fuels to determine whether biodiesel is a feasible alternative energy
source. Biodiesel is a possible alternative to traditional, non-renewable energy sources such as oil
and coal, having benefits such as being less toxic, having cleaner emissions, and being
renewable1. However, biodiesel also has drawbacks such as its higher viscosity which clogs fuel
tanks and its utilization of land that could be used for agriculture1. In this experiment there were
two parts: first, biodiesel was synthesized from vegetable oil. The chemical reaction of the
synthesis was (Glyceryl trilinoleate) + 3(Alcohol) -> 3 (Methyl linoleate) + (Glycerol). The
theoretical yield of biodiesel mass was found by converting volume of vegetable oil to moles,
then converting moles of vegetable oil to moles of methyl linoleate and finally converting moles
of methyl linoleate to grams. The percent yield of synthesis was found by (experimental yield of
biodiesel mass)/ (theoretical yield) x 100%. Percent yield was (15.52 g)/ (17.85 g) = 87.95%.
The biodiesel was then combusted, and its viscosity and enthalpy of combustion were measured.
Second, various biofuels were tested for toxicity through an eco-toxicity assay that measured the
germination of radish seeds against the concentration of biofuel. T will be compared with LD50
values found in literature, which give the amount of ingested sample that will kill 50% of the
sample4; the LD50 for diesel is 0.49 g/kg of rat3, and the LD50 of biodiesel is 5.001 g/kg of rat2.
The data will show the viscosity, enthalpy of combustion, and toxicity of the synthesized
biodiesel and other biofuels, and this information will be used to determine if biodiesel is a safe
and efficient alternative energy source.

Methods:
This experiment was taken from the Chemistry 1A Laboratory Manual1. Biodiesel was
synthesized from vegetable oil with 0.4 M NaOH by mixing the two substances in a centrifuge
tube, shaking the tube vigorously to mix them, and then allowing the substances to react for a
week. The reaction of 20 mL of vegetable oil and 5 mL of NaOH solution produced 17.5 mL of
biodiesel; the atom economy of the reaction was 70%. The synthesis reaction produces biodiesel
and glycerol, with trace amounts of impurities as well. To isolate the biodiesel sample, first the
glycerol layer that formed on top of the biodiesel was pipetted out. Then the biodiesel was
washed by the addition of 5 mL of 1 M NaCl solution, which drew water out from the biodiesel
sample and formed an aqueous layer that was then pipetted out. This salt wash also removed
water-soluble impurities. The sample was further dried by the addition of anhydrous MgSO4, a
solid that is insoluble in biodiesel, which absorbed lingering water molecules and then was
filtered out. After synthesis, the yield of reaction was determined by the following calculation:
(experimental yield of biodiesel mass) / theoretical yield x 100%. Theoretical yield of biodiesel
mass was calculated by converting volume of vegetable oil to moles, then converting moles of
vegetable oil to moles of methyl linoleate, then converting moles of methyl linoleate to grams;
the theoretical yield was 15.52 g. Experimental yield was found to be 17.85 g by measuring the
mass of the biodiesel sample in the centrifuge tube, and subtracting the mass of the empty tube.
Percent yield was found to be (15.52 g/17.85 g) x 100% = 87.95%.
The isolated biodiesel was then combusted and its heat of fusion was found through
calorimetry. Lab synthesized biodiesel was poured into a scintillation vial and a wick was soaked
in the fuel. The vial was covered with foil except for where the wick was exposed. The wick was
lighted, then the vial was placed underneath a soda-can calorimeter. The change in temperature
of the water in the calorimeter and the change in mass of the biofuel was measured for each trial.

This procedure was repeated for the other fuels. Combustion enthalpy was found through the
following calculation: Hcomb = qcal / g biodiesel.
Viscosity was measured by recording the time the solution took to pass through a Pasteur
pipette. The data showed that vegetable oil was the most viscous (taking 44.72 seconds), but
glycerol, which experimentally took 14.72 seconds, should have had the longest flow time and
highest viscosity. It was found that the lab synthesized biodiesel (8.90 sec) and commercial
biodiesel (5.50 sec) were both more viscous than methanol (1.00 sec) and diesel (2.96 sec).
For the eco-toxicity assay, biofuel solutions of different concentrations prepared through
serial dilution. First, 30 radish seeds were added to each of six petri dishes that were lined with
filter paper. Then, 20 mL of 10% (by weight volume) 2-butanol solution was obtained and 10 mL
of it was added to the first petri dish; 10 mL of deionized water was added to the remaining 10
mL of 10% solution to make the 5% solution, which was added to the second petri dish. This
method of adding 10 mL of water to the remaining 10 mL of previous solution was used to
prepare the 2.5%, 1.25%, and 0.625% solutions. Deionized water was added to the last petri dish
as the control. After one week, the number of germinated seeds were counted. Each lab group
performed the same procedures but used a different biofuel; class data about the percent of
germination for the different concentrations of different biofuels was compiled, and the relative
toxicities of the different biofuels was determined. Results showed that lab synthesized biodiesel
was the least toxic, as 95% of seeds germinated at 10% fuel concentration, followed by
commercial biodiesel (54.5% at 10% concentration) and methanol, ethanol, and 2-butanol (0%
germination at 10% concentration).

Results:
Table 1: Seed Germination for 2-Butanol Toxicity Assay by weight percent of biofuel
# of Seeds Germinated
% of Seeds Germinated

0%
27
90

0.625%
29
97

1.25%
27
90

2.5%
0
0

5%
0
0

10%
0
0

Table 1 presents the number and percent of seeds germinated after one week. Note that the three
lowest concentrations, 0%, 0.625%, and 1.25% yielded the greatest percentage of seeds
germinated and concentrations greater than or equal to 2.5% yielded no germination.
Graph 1: Eco-Toxicity Assay Results for Various Biofuels

Germination of Radish Seeds for Various Biofuels

Germination (%)

90
80
70
60
50
40
30
20
10
0

10

Concentration of Biofuel (% by weight volume)

Lab Biodiesel

Methanol

Ethanol

2-Butanol

Graph 1 presents the percent of germination for the different concentrations of various biofuels.
Lab biodiesel was the least toxic, as 95% of seeds germinated even at 10% concentration. 2Butanol was the most toxic, as germination dropped to 0% at 2.5% concentration and higher.

Table 2: Viscosity Measurements

Time (sec)

Commercial Biodiesel

5.50

Methanol

1.00

Glycerol (pure sample)

14.22

Glycerol layer (from experimental synthesis)

1.88

Vegetable Oil

44.72

Diesel

2.96

Lab Synthesized Biodiesel

8.90

Table 2 (above) shows the viscosities for various biofuels and glycerol. Vegetable oil had the
highest viscosity (44.72 sec). Diesel had a low viscosity (2.96 sec). Experimental biodiesel (8.90
sec) was about 3 times as viscous as the commercial biodiesel. This higher viscosity of could be
attributed to imperfect isolation of the fuel, which left high-viscosity glycerol in the sample.
Table 3: Ethanol Combustion
Trial

Initial
Mass (g)

Final
Mass (g)

Initial
Final
mass
T
Ccal
Temp
Temp
(kj/C)
(C)
(C)
1
17.2030
16.5583
24.0
33.5
0.6447
9.5
2.0146
2
16.4867
12.8998
23.5
33.5
0.5869
10.0
1.743
3
15.8905
15.2727
23.0
33.5
0.6178
10.5
1.747
Table 3 shows the results of the three trials of ethanol combustion that were used to calculate
Ccal. Ccal was found through the following calculations:
1) mol ethanol = g ethanol x (mol ethanol/molar mass of ethanol)
2) qcal= Hcomb(mol of ethanol)
3) Ccal=qcal/T
For trial 1, molethanol=0.6447 x (mol ethanol/46.07 g) = 0.01399 mol.
qcal=1368 kJ/mol (0.01399 mol) = 19.138 kJ. Ccal= 19.138 kJ/9.5 C = 2.0146.
The average Ccal was found to be 1.835 kJ/C.
Table 4: Biodiesel Combustion for Synthesized Biodiesel

Trial
1
2
3

Initial
Mass (g)
18.8667
18.3793
17.606

Final
Mass (g)
18.3793
17.8501
17.5699

Initial
Temp (C)
22.5
22.0
24.0

Final
Temp (C)
32.0
31.0
27.0

mass

0.4874
0.5292
0.1907

9.5
9.0
3.0

Hcomb
(kJ/g)
-35.966
-31.207
-28.867

Table 4 summarizes the data collected during three trials of synthesized biodiesel combustion.
Enthalpy of Combustion was found using the following calculation: Hcomb = qcal/gbiodiesel. Heat of
the calorimeter was found using the following calculation: qcal= T x Ccal, which was determined
to be 1.835 kJ/C. Average Hcomb was -29.647 kJ/g. Note that Hcomb is a negative number,
meaning that the reaction gives off heat.
Table 5: Heats of Combustion for Vegetable Oil, Biodiesel, and Diesel
Fuel

Initial
Final
Mass (g) Mass
(g)
20.46
20.28
20.51
20.10

Initial
Temp
(C)
22
20

Final
Temp
(C)
26
30

mass

Hcomb
(kJ/g)

Vegetable Oil
0.18
4
-33.7
Commercial
0.41
10
-34.6
Biodiesel
Diesel
18.44
18.09
21
30
0.35
9
-46.7
Table 5 summarizes the data collected for the combustion of vegetable oil, commercial biodiesel,
and diesel. Note that the heat of combustion for all three fuels is greater than the average heat of
combustion for the lab synthesized biofuel (-29.647 kJ/g). This shows that the lab synthesized
biodiesel was a less efficient fuel. Diesel was the most efficient, with a Hcomb of -46.7 kJ/g, and
commercial biodiesel was second-most efficient with a Hcomb of -34.6 kJ/g. Discussion and
Conclusions:
The results show that the lab synthesized (LS) biodiesel had advantages and
disadvantages over other biofuels and diesel: it was the least toxic, but it was also the least
efficient and one of the most viscous. Graph 1 shows lab synthesized biodiesel was the least
toxic of the fuels tested; at 10% concentration by weight volume 95% of the seeds germinated,
versus 54.5% of seeds for commercial biodiesel and 0% for 2-butanol, methanol, and ethanol.

Also, Graph 1 shows that the LD50 for commercial biodiesel was about 10% by weight volume,
as only 54.5% of the seeds germinated. This data does not concur with literature; the LD50 (the
amount of sample that will kill 50% of the test sample) of biodiesel found by Wil Research
Laboratories, Inc.2 is 5.001 g/kg, in comparison to 6.48 g/kg for 2-butanol7; the data should have
shown that 2-butanol was less toxic than biodiesel. Biodiesel is definitively less toxic than diesel,
whose LD50 was found to be 0.49g/kg 3. One possible source of error is that only one assay was
prepared for each type of fuel, so there was not extensive data from which to draw conclusions.
Future versions of this experiment should include more trials of the toxicity assay.
In terms of fuel efficiency, as seen in Tables 4 and 5 the average heat of combustion for
LS biodiesel was -29.47 kJ/g, versus -34.6 kJ/g of commercial biodiesel and -46.7 kJ/g for diesel.
Fuels with higher heats of combustion are more efficient because they produce more energy per
gram of fuel6. The data shows that diesel was about 59% more efficient than LS biodiesel and
about 35% more efficient than commercial biodiesel. One possible source of error during the
combustion of LS biodiesel was that the flame kept dying down, so heat from combustion went
into heating the foil covering the vial instead of the calorimeter, resulting in a lower temperature
change and thus a lower calculated heat of combustion. In addition, the calorimeter used was not
a completely closed system, so some heat from the combustion reaction escaped into the air. To
improve accuracy, a more precise apparatus such as a bomb calorimeter should be used in future
versions of this experiment.
For viscosity, Table 2 shows that LS biodiesel was the most viscous out of commercial
biodiesel and diesel; LS biodiesel took 8.90 seconds to travel through the Pasteur pipette,
commercial biodiesel took 5.50 seconds, and diesel was the least viscous, taking 2.96 seconds.
One factor contributing to error could be that the LS biodiesel was not perfectly isolated before it
was tested for viscosity; glycerol, a waste product of the synthesis reaction, has a high viscosity

and its presence in the LS biodiesel sample could have increased its viscosity. In addition, the
method of measuring viscosity used was prone to error because it relied on how fast the observer
could recognize when the fuel finished flowing and how accurately they could operate a
stopwatch. Because many of the flow times occurred in seconds, even a small error on the
observers part could greatly change the data.
In conclusion, biodiesel is a feasible alternative energy source because it is less toxic than
other biofuels and diesel and has a comparable heat of combustion. Other potential advantages of
biodiesel are that it is potentially better for the environment because it is produced from
renewable resources such as vegetable oils, animal fats, and cooking oils1 and that biodiesel has
cleaner emission because it yields 3.2 units of fuel productivity for every 1 unit of fossil energy
consumed, whereas diesel yields just 1.2 units1. However, further research is needed to increase
the heat of combustion of biodiesel and to find ways of lowering its viscosity, in order to surpass
diesel in these two categories.
References:
[1] Laboratory Manual: Chemistry 1A Fall 2010. Department of Chemistry; University of
California, Berkeley. Regents of the University of California, 2010.
[2] "Bio-Diesel." Bio-Diesel. Green-Trust, n.d. Web. 10 Nov. 2015.
[3] Safety Data Sheet. Woodbridge, NJ: Hess Corporation, 30 Aug. 2012. PDF.
[4] "OSH Answers Fact Sheets." Government of Canada, Canadian Centre for Occupational
Health and Safety. Canadian Centre for Occupational Health and Safety, n.d. Web. 13
Nov. 2015.
[5] Material Safety Data Sheet Biodiesel. San Antonio, TX: Tesoro, n.d. PDF.
[6] "Fuel Definitions." Fuel Definitions. Aus-e-tute, n.d. Web. 13 Nov. 2015.