FDE-226

FOOD ANALYSIS LABORATORY

I PLEDGE THAT I HAVE PREPARED ALL REPORT BY MYSELF, RELYING ON
SOLELY ON MY OWN

Name & Surname: Ada Uçar

Student ID: 20210709023

EXPERIMENT-5
Determination of Fat Content

1. INTRODUCTION

The accurate determination of the fat content in food samples is important for various purposes,
such as declaring nutritional value, ensuring quality control, and complying with labeling
regulations. ( Borodulin, et al ,2020).
The Soxhlet method which is a typical technique for the extraction of analytes from solid materials,
is an extensively used method for determining fat content. This approach has been used regularly
in analytical laboratories since its development in 1879. It is also frequently used to calculate the
fat content of a variety of foods, including cereals, dairy products, meat, and oils. This approach
includes continuous extraction using a solvent (often hexane) until a steady state is reached. This
method is still a common procedure and is frequently used as a standard for how well modern
extraction techniques operate. ( Borodulin et al ,2020)
The Soxhlet Extraction Method has several Advantages, Disadvantages and Limitations. Some of
the significant features are listed below

Advantages:
• Soxhlet extraction is a reliable and efficient method for extracting lipids from samples with
low-to-moderate lipid content, such as nuts.
• Soxhlet extraction can extract fat from complex matrices, such as those found in pistachios,
with minimal interference from other compounds.
• Soxhlet extraction is a well-established method and is widely used in lipid analysis.
• The method is relatively inexpensive and requires simple equipment.

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(Basha & Rajagopal ,2009).
Disadvantages:
• Soxhlet extraction is time-consuming, as the process can take up to several hours.
• The method requires a large amount of solvent, which can be expensive and hazardous to
handle.
• Soxhlet extraction can lead to thermal degradation of the extracted lipids due to prolonged
exposure to hot temperatures.
• The method may extract other compounds along with the lipids, leading to potential
interference with downstream analysis.
(Barbieri, & Kennicutt II, 2006)

Limitations:
• Soxhlet extraction is not suitable for high-throughput analysis due to its time-consuming
nature.
• The method is not effective for samples with high lipid content, as the extracted lipids can
saturate the solvent and lead to reduced efficiency.
• Soxhlet extraction may not be suitable for heat-sensitive samples, as the hot temperatures
used in the process can lead to thermal degradation.
(Basha & Rajagopal, 2009).

While there are alternative methods available for fat extraction, Soxhlet extraction remains a
popular and reliable method for extracting lipids from pistachios and other samples with moderate
or high lipid content. The advantages and limitations of the method must be carefully considered
when choosing an appropriate extraction technique for specific research. (Proestos & Komaitis ,
2005)
Alternative methods to Soxhlet extraction are:
Solvent extraction: This method involves directly extracting lipids using a solvent, which is less
time-consuming than Soxhlet extraction but can lead to more interference from other compounds.
(Kocadağlı & Özdemir ,2017)
Supercritical fluid extraction: This method uses supercritical CO2 to extract lipids, which is faster
and more selective than Soxhlet extraction but requires more expensive equipment. (Kocadağlı &
Özdemir, 2017)
Microwave-assisted extraction: This method involves using microwaves to heat the sample and
solvent, which can reduce extraction time compared to Soxhlet extraction but may lead to more
interference from other compounds. (Eskilsson, C. S., & Björklund ,2000).

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Solvent extraction methods (Soxhlet, Goldfish, and Mojonnier) involve using solvents to extract
lipids from the sample. These methods are simple to perform but can be time-consuming and
require large sample sizes. (Proestos, & Komaitis , 2005).
Nonsolvent extraction methods (Babcock) rely on chemical reactions to estimate the fat content.
The Babcock method is useful for determining the fat content in milk and dairy products. (Guo, &
Guilfoil, 2021).
Instrumental approaches (near-infrared spectroscopy, gas chromatography) depend on the physical
and chemical properties of lipids to estimate fat content. These methods are faster and require
smaller sample sizes than traditional extraction methods but may require more expensive
equipment and specialized training to perform. (Guo & Guilfoil, 2021)
The purpose of this laboratory exercise is to determine the pistachio's lipid content. The reason
Soxhlet extraction is used is because this method is suitable where the desired compound has a
low solubility in a solvent, and unwanted side product is insoluble in that solvent. A
straightforward filtering would be used if desired content was highly soluble in a solvent. (S.
Kumar, et al 2018)
Also, another advantage of Soxhlet extractor is that it has a reducing effect on the amount of
solvent and wet sample used, because the target component is being extracted almost at its maximal
rate. ( Borodulin et al, 2020)
The choice of method depends on the specific application, available equipment, and desired level
of accuracy and precision.

2. MATERIALS AND METHODS

2.1. Materials

250 ml hexane (Sigma-Aldrich)

Pistachio sample (Peyman) obtained from Burçak Gıda and stored at room temperature until
analysis.
Flask, Soxhlet extractor, tong, rotary evaporator, hot plate, thimble, gloves.

2.2. Methods
The Pistachio sample was pre dried using a vacuum oven at 70ºC, 635 mmHg, for 24 h.
(The fat content was recorded as written on the label so it can be compared afterwards)

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30 g of pistachio sample is grinded slightly using a mortar and pestle
After the weight of the thimble is tared 2 grams of sample was put into thimble and weighed
The boiling flask was weighed and labeled then placed in a Soxhlet hot plate. 250 ml of hexane
(Sigma-Aldrich) was put in each flask and extracted for 6 h.
The flask was removed from the Soxhlet extractor using tongs, The flask was cooled to room
temperature, and the remaining hexane in the flask was evaporated using a rotary evaporator. After
all hexane is evaporated, the flask containing the extracted fat is weighed and required calculation
was made.

(𝐵𝑜𝑖𝑙𝑖𝑛𝑔 𝑓𝑙𝑎𝑠𝑘 𝑓𝑖𝑛𝑎𝑙(𝐹𝑎𝑡+𝐹𝑙𝑎𝑠𝑘)(𝑔)− 𝐵𝑜𝑖𝑙𝑖𝑛𝑔 𝑓𝑙𝑎𝑠𝑘 𝑖𝑛𝑖𝑡𝑎𝑙 (𝑔))

%Fat = ⋅ 100 (Eq.1)
Pr 𝑒 𝑑𝑟𝑖𝑒𝑑 𝑆𝑎𝑚𝑝𝑙𝑒 (𝑔)

3. RESULTS
Table 1. Data from Soxhlet Extraction
Replicates Pre-dried Boiling Boiling flask-final
sample (g) flask- (Fat+Flask)(g)
initial (g)
Replicate 1 2.11 204.33 205.64

Replicate 2 2.10 212.24 213.60

Replicate 3 2.08 212.75 213.67

205.64 −204.33
% Fat of Replicate 1 = ⋅ 100 = % 62.09
2.11
213.60 −212.24
% Fat of Replicate 2 = ⋅ 100 = % 64.76
2.10

213.67 −212.75
% Fat of Replicate 3 = ⋅ 100 = % 44.23
2.08

The results above were calculated using (Eq.1)

X(mean) = % 57.03
STD = 11.26
Results = 57.03 ± 11.26

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Label Value of Pistachio Fat = %44.4
(44.4 grams of fat per 100 grams pistachio product)

4. DISCUSSION

The determination of fat content in food products is significant in the food industry as it provides
important information for labeling, quality control, and product development. Labeling the fat
content of a food product is important for consumer awareness and can impact their decisions
when purchasing a product. Recording the fat content during production can ensure consistent
quality and help prevent any deviations or issues with the product. (Ramaswamy & Sivakumar ,
2014)
The results of the Soxhlet method indicate that the pistachio sample has a high fat content, with a
mean value of 57.03% and a standard deviation of 11.26 (57.03 ± 11.26). This indicates that the
sample contains more than the expected fat content of 44.4 grams per 100 grams of the product,
as stated on the label. The high difference between the measured fat content and the expected fat
content could be due to several factors, including variations in the sample, experimental errors, or
inaccurate labeling. (Kocadağlı, & Özdemir, 2017)
Comparing the results with literature values of pistachio samples gathered around turkey ranging
from 47.65 % to 63.31% it is safe to conclude that values are within the expected range with a slight overflow
in Replicate 3 where result was 64.27 percent fat. (Satil, F., Azcan, N., & Başer, K. H. C., 2003)

Factors affecting the method's performance can include equipment calibration, chemical purity,
operator technique, and sample preparation errors. Statistical measures such as mean and standard
deviation can be used to evaluate the method's performance and identify any potential sources of
error or variability. In this case, the high standard deviation suggests that there may be some
variability in the sample or the method, which could be addressed by improving the method or
optimizing the sample preparation. (Kocadağlı, T., & Özdemir, N. ,2017)
In conclusion, the Soxhlet method can provide a reliable and accurate measurement of fat content
in food products. However, it is important to consider potential sources of error and variability and
to compare the results with expected values or standards to ensure accuracy and consistency.

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Kumar, S., Bhandari, C., Sharma, P., & Agnihotri, N. (2018). Role of Piperine in Chemoresistance.
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