Andre Smith

October 21, 2015

FST 101A-A06, Wednesday 9 AM
RH
LAB 4 Total Acidity of Olive Oils
I. Purpose/Objective
The purpose of this lab was to look at the free fatty acid content of three different
types of olive oils, extra virgin olive oil, pure olive oil, and extra light olive oil by
titration with sodium hydroxide.
II. Introduction
Olive oil contains a mixture of triglycerides and free fatty acids. A triglyceride
consists of a glycerol backbone with three fatty acids attached. Olives contain a
variety of fatty acids, but mostly contain oleic acid. The fatty acids can be cleaved by
enzymes such as lipase and as olives begin to ripen, lipase naturally acts on the
triglycerides in the olives which increases the free fatty acid content. Through
titration, the %acidity of the olive oil can be tested to determine the quality of the
olive oil. Often the free fatty acid content of olive oil is considered to be the % oleic
acid as it makes up over 55% of the fatty acid profile.
Extra Virgin Olive Oil (EVOO) is olive oil that is bottled after being cold pressed
once. There is no chemical or heat treatment, so it has the lowest free fatty acid
content and the highest polyphenol content and is considered to be the highest quality
of the different olive oil types. Pure Olive Oil (POO) has a slightly higher free fatty
acid content. EVOO has under 0.8% free fatty acid content while POO has about 1-
2%. POO also has a lower polyphenol content. POO is often a mix of different quality
olive oils. Extra Light Olive Oil (ELOO), also known as Refined Olive Oil, has the
lowest free fatty acid content and the lowest polyphenol content. Typically a lower
free fatty acid content means that the olive oil is a higher quality, but in the case of
ELOO the low percentage of free fatty acid is due to neutralization of the acids in the
refining of the oil.
Each different class of oil has its own attributes. EVOO has strict standards and
cannot be sold in the US unless those standards are met. Each type of olive oil has a
 different use and knowing what type of olive oil is being purchased is important for
carrying out its intended use such as cooking or tasting.

III. Procedure
The procedure for Lab 4 is found in Principles of Food Composition, Laboratory
Manual, FS&T 101A (2015) page 31. There were no modifications for this lab.
IV. Data
Table 1: Extra Virgin Olive Oil Titration. Normality of NaOH = 0.1104 N
Titration # Weight (g) Start NaOH End NaOH Total NaOH
Volume (ml) Volume (ml) used (ml)
1 27.5 19.10 22.10 3.00
2 27.6 22.21 25.40 3.19
3 27.6 22.50 28.20 2.70

Table 2: Extra Light Olive Oil Titration. Normality of NaOH = 0.1104 N

Titration # Weight (g) Start NaOH End NaOH Total NaOH
Volume (ml) Volume (ml) used (ml)
1 27.9 28.35 29.70 1.35
2 27.9 29.70 30.80 1.10
3 28.0 30.95 32.00 1.05

Table 3: Pure Olive Oil Titration. Normality of NaOH = 0.1104 N

Titration # Weight (g) Start NaOH End NaOH Total NaOH
Volume (ml) Volume (ml) used (ml)
1 28.6 32.10 32.90 0.80
2 28.7 33.00 34.05 1.05
3 28.6 34.19 35.05 0.96
 Table 4: %Acidity, mean, and standard deviation for each titration.

EVOO %Acidity POO %Acidity ELOO %Acidity

1 0.336 1 0.151 1 0.087

2 0.360 2 0.123 2 0.114

3 0.305 3 0.117 3 0.105

Average 0.333 Average 0.130 Average 0.102

Std Dev 0.028 Std Dev 0.018 Std Dev 0.014

Table 5: Sensory evaluation of olive oils

Oil # Color Depth Aroma Depth Sourness/Acidity Fruitiness/Nuttiness Guess
1 Lightest Weakest Least Least POO
2 Middle Middle Middle Middle ELOO
3 Darkest Strongest Most Most EVOO

Table 6: ANOVA Single Factor for % acidities of each titration.

Anova: Single Factor

SUMMARY
Groups Count Sum Average Variance
Column 1 3 1.001 0.333667 0.00076
Column 2 3 0.391 0.130333 0.000329
Column 3 3 0.306 0.102 0.000189

ANOVA
Source of VariationSS df MS F P-value F crit
Between Groups
0.095817 2 0.047908 112.4022 1.76E-05 5.143253
Within Groups
0.002557 6 0.000426

Total 0.098374 8
 Average % Acidity
0.4

0.35

0.3

0.25

0.2

0.15

0.1

0.05

0
EVOO POO ELOO

Fig. 1: Average % acidity of each oil with standard deviation.

V. Calculations
Calculating average % acidity:
Using Group Nines data for the EVOO %acidity, calculating the mean of the trials can
be done by adding all 3 numbers together and dividing by the sample size.
0.336% + 0.360% + 0.305% = 0.333%
3
Calculating sample standard deviation:

Once the mean has been calculated, the standard deviation can begin to be calculated by
subtracting the mean from each number in the data set.
0.336-0.333= 0.003
0.360-0.333= -0.027
0.350-0.333= -0.017
Square each of the differences
(0.003)2 = 9.00 x 10-6
(0.027)2 = 7.29 x 10-4
 (0.017)2 = 2.89 x 10-4
Add the squared numbers together and divide by n-1. In this case, 3-1=2. Then, take the
square root of the resulting number to calculate sample standard deviation.
9.00 x 10-6 +7.29 x 10-4 +2.89 x 10-4 = 1.03 x 10-3
2
5.1510-4= 0.023

Calculating %Acidity
First, multiply the volume of NaOH used in liters times the concentration of the NaOH.
This gives the equivalents of NaOH. Oleic is a monoprotic acid, so the equivalents of NaOH is
the same as the equivalents of oleic acid. Multiply the equivalents of oleic acid times the
equivalent weight of oleic acid (282 g/mol) to find the mass of oleic acid.
For the first titration of EVOO:

0.003 L NaOH x 0.1104 N NaOH x 282 g/mol = 0.0933g oleic acid

Next, divide the mass of oleic acid by the mass of the olive oil used. Multiply this
number by 100 to get the % acidity.

(0.0933 g oleic acid/ 27.5 g olive oil) x100 = 0.336%

VI. Results/Discussion
Table 6 shows that at least one of the oils is significantly different from the other oils
because the F value is greater than the F critical and the P value is lower than the alpha
value. According to figure 1, EVOO is significantly different from POO and ELOO. The
error bars on ELOO and POO do overlap so they are not significantly different. To be
labeled as EVOO the oil must abide by a certain set of standards. EVOO is not heat or
chemically treated so it has a low fatty acid content and high polyphenol content.
However, in this experiment it was found that both ELOO and POO have lower fatty acid
contents than the EVOO. Refined oils are sometimes neutralized and this can be seen
 with the low % acidity for each titration of ELOO. POO is a mixture of different quality
olive oils. The low fatty acid content shows that refined oils can be part of this mixture.
In the taste test, EVOO was assumed to be olive oil #3. It had the darkest color, and
deepest aroma and taste. This experiment does not quantify polyphenol content, but
perhaps the rich attributes are contributed by the minimal processing of the olives to
make EVOO. Olive oil #2 was assumed to be POO, because the flavor and aroma profiles
were much less intense than oil # 3. Oil # 1 had even weaker qualities and was assumed
to be ELOO. While it was difficult to sense acidity in the olive oils by taste, the titration
data shows that all of the oils had less than 0.4% acidity. Pure olive oil should have a
%acidity closer to 1%, but only displayed an average of 0.130%, which may contribute to
the difficulty in tasting different levels of acidity in these oils.

VII. Conclusion
The purpose of this experiment was to quantify the % oleic acid of each of the three olive
oils and see if each oil fit the qualifications for its particular grade. % Acidity is an
important factor in determining a grade of oil along with its processing method and
polyphenol content.

VIII. Questions
1. Describe in your own words what extra virgin olive oil is. Use both analytical and
sensory terms/ attributes in your description.
EVOO is the highest quality olive oil. It has a specific set of standards it must abide by to
be labeled and sold as EVOO. It should have a dark green color and high polyphenol
content. It should also have a free fatty acid content of <0.8%. It is able to maintain its
flavor components due to its minimal processing. The olives are harvested early and cold
pressed once. There is no heat or chemical treatment on the olives.
2. Based on your statistical analysis are there any real differences in % oleic acid content
among the oils? In your answer make sure you refer to F value and post hoc analysis.
In the ANOVA single factor analysis, the F value was larger than the F critical value and
the P value was lower than the alpha value of 0.05 indicating there at least one group was
 significantly different from the others. To determine which groups are different, a post
hoc analysis must be done. Excel cannot provide this test, so the mean %acidity of each
of the oils was graphed with their standard deviations. The error bars for POO and ELOO
overlapped, so they are not significantly different but the EVOOs error did not overlap
anything indicating that group is significantly different from the other ones.
3. What assumptions are you making by basing the titratable acidity on oleic acid molecular
weight? Can you suggest a better alternative?
By using the MW of oleic acid to calculate % acidity, we are assuming the most of the
fatty acids in the oil are oleic acid. Oleic acid can make up up to 80% of the fatty acid
profile in olive oil. A more accurate result can come from the analysis of other acids in
the solution such as palmitic or linoleic.