EXTRACTION OF OIL FROM PUMPKIN SEED USING

DIFFERENT EXTRACTION METHODS AND

DETERMINATION OF YIELD AND QUALITY. 
 
 
 
 
 
Authored
by

Israel Chumba BSIC/2020/90574 

Edwin Sirengo BSIC/2020/90246 
 
 
 
 
 

Project proposal submitted to the natural sciences department as a

Partial requirement for Mount Kenya university's Bachelor of
Science in industrial chemistry degree award.

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 I. DECLARATION 
We declare that this proposal is our work and has not been submitted by anybody else in any
other university for the award of any degree to the best of our knowledge. 

  EDWIN SIRENGO BSIC/2020/90266

Sign………………………………… Date………………………………………. 

ISRAEL CHUMBA BSIC/2020/90574

Sign………………………………… Date………………………………………. 

Department of Natural science 

Mount Kenya University 

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 II. CERTIFICATION 
  
This project has been submitted for examination with my approval as the candidate’s supervisor. 
  
Sign…………………………….                  Date ……………………………………………… 
Peter kauri
[Designation] 
Natural science
  

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 III. Acknowledgements 
We are grateful to Peter Karuri for guiding us throughout our research.
 

iv
 IV. Abstract 
This research aims to extract pumpkin seed oil using different methods and examine its yield and
quality. Data will be collected by measuring the acid value and edibility of the oil and
determining its iodine value. The expected results are to provide an accurate estimation of the
oil's quality and yield. The samples of pumpkin seed oil will be collected and analyzed using
various methods. These methods include measuring the acid value, which measures how much
acidic material is present in the oil, and determining the iodine value, which measures the
amount of unsaturated fatty acids in the oil. The results of these experiments will provide a better
understanding of the oil's quality and yield. Additionally, the research will provide insight into
the optimal conditions for extracting pumpkin seed oil and allow for better control and
optimization of the process. Ultimately, the data collected and analyzed in this project will
provide valuable information on the quality and yield of pumpkin seed oil and enable more
efficient product production.

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 V. TABLE OF CONTENT 

Contents
I. DECLARATION........................................................................................................................ii
II. CERTIFICATION..................................................................................................................iii
III. Acknowledgements................................................................................................................iv
IV. Abstract....................................................................................................................................v
V. TABLE OF CONTENT..........................................................................................................vi
VI. ACRONYMS AND ABBREVIATION..............................................................................viii
VII. List of tables..........................................................................................................................ix
CHAPTER 1...................................................................................................................................1
Introduction....................................................................................................................................1
1.1 Background information.........................................................................................................1
1.2 Motivation and problem statement........................................................................................2
1.3 Research identification............................................................................................................2
1.4 RESEARCH OBJECTIVES...................................................................................................2
1.4.1 General objectives.................................................................................................................2
1.4.2 Specific objectives...........................................................................................................2
3.4 Hypothesis........................................................................................................................2
CHAPTER 2...................................................................................................................................3
2. Literature review.......................................................................................................................3
2.3 IMPORTANCE TO HUMAN HEALTH..............................................................................4
CHAPTER 3...................................................................................................................................5
RESEARCH METHODOLOGY.................................................................................................5
3.1 Study area........................................................................................................................5
3.2 Research design........................................................................................................................5
Methodology...................................................................................................................................5
3.3 Sample and Sample Preparation............................................................................................5
3.4 SOXHLET EXTRACTION....................................................................................................6
PROCEDURE................................................................................................................................6
3.5 COLD PRESSING...................................................................................................................7
3.5 ANALYSIS OF THE OIL EXTRACTED....................................................................7
3.5.1 Determination of Oil Yield.............................................................................................7

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3.6.2 IODINE VALUE...................................................................................................................8
Procedure........................................................................................................................................8
3.6.3 ACID VALUE.......................................................................................................................9
DATA ANALYSIS.......................................................................................................................10
Appendix.......................................................................................................................................11
Budget 11
table 1.1 1. laboratory utilities, resources, and cost estimating...............................................11
Work plan.....................................................................................................................................11
table 1.1 2. activity work and period to be done.......................................................................11
4. References......................................................................................................................12

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 VI. ACRONYMS AND ABBREVIATION 
PSO- Pumpkin seed oil 
LDL- Low-density lipoprotein
SFE- Supercritical fluid extraction
PA- palmitic Acid
OA- Oleic Acid

viii
 VII. List of tables
table 1.1 1. laboratory utilities, resources, and cost estimated................................................11
table 1.1 2. activity work and period to be done.......................................................................11

 
 
 

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 CHAPTER 1 

Introduction 

1.1 Background information 

Pumpkin is a widely grown crop of high economic importance belonging to the family
Cucurbitaceae (GEMROT et al.; DURANTE et al.,2014). Its seeds are a rich source of
nutritionally valuable oil. Pumpkin seed oil (PSO) is a deep green oil obtained by cold pressing
the hulled pumpkin seeds. It is increasingly used as a base oil in cosmetic and food products
(Gangwar et al., 2017). PSO has gained significant attention due to its high nutritional value and
health benefits. It is a rich source of unsaturated fatty acids, minerals, vitamins, and phytosterols
(Kaur et al., 2018). The fatty acid composition of PSO is dominated by linoleic Acid (LA), and it
also has high concentrations of oleic Acid (OA) and palmitic Acid (PA) (Sarker et al., 2020).
The high LA and OA content mainly gives PSO unique properties such as high oxidative
stability and good spreadability (Hajebrahimi et al., 2017).
In order to obtain the oil from pumpkin seeds, various extraction methods can be employed. The
most commonly used methods are mechanical pressing, solvent extraction, and supercritical fluid
extraction (SFE). Mechanical pressing is the simplest method and involves using low
temperatures and low pressures to extract the oil from the seeds. Solvent extraction involves
using solvents such as hexane and is the most efficient method for oil extraction. SFE is a
relatively new method and involves the use of supercritical carbon dioxide as the extracting
agent. The quality of the extracted oil depends on the method used for extraction. It is essential
to measure the acid value, iodine value, and edibility of the oil in order to determine its quality.
The acid value is a measure of the amount of free fatty acids present in the oil and is an
indication of the oil's oxidative stability. The iodine value is a measure of the level of
unsaturation in the oil and is an indication of the oil's spreadability. The edibility of the oil is
determined by its taste and smell. The yield of the extracted oil also depends on the method used
for extraction. The yield is the percentage of oil obtained from the raw material, indicating the
extraction process's efficiency. This research aims to extract oil from pumpkin seeds using
different extraction methods and determine the quality and yield of the extracted oil. The oil's

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 Acid value, iodine value, edibility, and yield will be measured to determine the extraction
methods' quality and efficiency.

1.2 Motivation and problem statement 

This research aims to investigate the most efficient and economical method for extracting oil
from pumpkin seeds to produce high-quality oil with good edibility and unsaturation levels. The
goal is to identify the best extraction method that yields the most oil cost-effectively while also
considering any health, environmental, and economic issues associated with the process. The
results from this research will provide valuable information for potential commercial
applications of pumpkin seed oil, such as in the food and cosmetics industries (Hussain et al.,
2020).

1.3 Research identification 

The primary objective of this research is to compare the yield and quality of the oil obtained
from two different extraction methods of pumpkin seed oil. Additionally, this research seeks to
analyze the impact of various parameters on the yield and quality of the oil extracted from
pumpkin seeds and to identify the most effective extraction method. Is achievable through the
following objectives

1.4 RESEARCH OBJECTIVES 

1.4.1 General objectives 

This research aims to extract oil from pumpkin seeds using different methods and determine
yield and quality. 

1.4.2 Specific objectives

1 Determination of lipid yield from pumpkin seeds

2. To measure the acid value and edibility of the lipid extract obtained from two different extraction
methods 
3. To assess the iodine value and the level of unsaturation of the pumpkin seed oil obtained from
two different extraction methods. 

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 3.4 Hypothesis
The hypothesis is that pumpkin seeds can be a source of edible oil that can be extracted through
different techniques. These techniques can result in differences in the oil yield, acidity, edibility,
iodine values, and level of unsaturation produced.

CHAPTER 2 

2. Literature review  
Pumpkin seed oil (PSO) is a highly nutritious oil extracted from the seeds of pumpkin plants. It
has a unique nutty flavour and is often used in cooking. In recent years, the health benefits of this
oil have generated significant interest among researchers. Several extraction techniques have
been used to prepare PSO for consumption. Cold pressing is the most common method used to
extract pumpkin seed oil, as it is believed to be the best way to preserve the oil's nutritional
benefits. In this method, the seeds are ground up and pressed in a cold press to extract the oil
from the seeds. Other methods used to extract PSO include solvent extraction and supercritical
fluid extraction. The health benefits associated with PSO are numerous.
Studies have shown that the oil is rich in polyunsaturated fatty acids, which can help to reduce
cholesterol levels, improve heart health, and reduce inflammation. Additionally, PSO is a good
source of vitamins, minerals, and antioxidants, which can help to boost the immune system and
protect against chronic diseases. Despite the numerous health benefits associated with PSO,
some research areas still need to be explored. For instance, more studies are needed to determine
the optimal extraction method for preserving the nutritional benefits of PSO. Additionally,
research is needed to determine the ideal concentration of PSO for optimal health benefits.
Finally, further research is needed to determine the long-term health effects of consuming PSO
regularly.
In conclusion, pumpkin seed oil is highly nutritious with various potential health benefits.
However, more research is needed to determine the optimal extraction technique and the long-
term health effects of consuming PSO regularly. By addressing these research gaps, we can
further understand the health benefits of this oil and how best to utilize it for optimal health. A
systematic review of 16 studies published in Nutrition Research in 2016 found that pumpkin seed
oil supplementation may reduce total cholesterol, LDL, and triglyceride levels. The review also
suggested that pumpkin seed oil supplementation may reduce systolic blood pressure. A study

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published in the journal Phyto-therapy Research in 2017 examined the effects of pumpkin seed
oil on markers of cardiovascular risk in postmenopausal women. The study found that
supplementation with pumpkin seed oil significantly reduced markers of inflammation and
oxidative stress, as well as total cholesterol, LDL, and triglyceride levels. Several animal studies
have also investigated the potential cardioprotective effects of pumpkin seed oil. A study
published in Nutrition Research in 2017 found that supplementing rats with pumpkin seed oil
reduced their total cholesterol and LDL cholesterol levels.
Additionally, the study found that pumpkin seed oil supplementation reduced markers of
oxidative stress and inflammation. Overall, the evidence suggests that pumpkin seed oil
supplementation may benefit cardiovascular health by reducing cholesterol levels, blood
pressure, and markers of inflammation and oxidative stress. More research is needed to confirm
these findings in humans.
  

2.3 IMPORTANCE TO HUMAN HEALTH 

Pumpkin seed oil is a rich source of antioxidants, compounds that can help protect against cell
damage caused by free radicals. Free radicals are molecules that can cause damage to cells,
tissues, and organs and are associated with age-related diseases, such as heart disease, cancer,
and Alzheimer's. Antioxidants can help neutralize free radicals, which in turn can help reduce the
risk of age-related diseases. Pumpkin seed oil is particularly rich in carotenoids, including beta-
carotene, a powerful antioxidant. Research suggests that pumpkin seed oil can help reduce
inflammation, reduce oxidative stress, and protect against cardiovascular disease. It may also
reduce the risk of certain cancers, such as breast cancer. In addition to its antioxidant benefits,
pumpkin seed oil is a good source of essential fatty acids, including omega-3, which is essential
for a healthy heart.
  

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 CHAPTER 3 

RESEARCH METHODOLOGY 

3.1 Study area

The seeds will be collected from Thika's local market. This research will be conducted in the
industrial chemistry laboratory at Mount Kenya university. The laboratory is equipped with all
the necessary equipment and tools required to carry out the research.

3.2 Research design  

The research will use a quantitative approach to examine the quality and yield of pumpkin seed
oil. In order to limit the scope of the study, the following parameters will be considered:
- The type of pumpkin seed to be used.  
- The extraction method. 
- The yield (in terms of oil).  
- The acid value and edibility of the lipid extract. 
- The iodine value and level of unsaturation. 
Data will be collected through the following methods:  
- Visual inspection of the pumpkin seed. 
- Extraction of the oil using two different methods. 
- Analyzing acid value and edibility through titration and taste tests.  
- Analyzing iodine value and level of unsaturation through titration. 
The methods used in this study are justified in that they are suitable for analyzing lipid content
and will provide accurate and reliable results. (Huang et al., 2019; Aregheore, 2014).  

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 Methodology 

3.3 Sample and Sample Preparation 

The collected samples of pumpkin seeds will be cleaned by removing dust, debris, and other
impurities. The samples will then be dried in the sun for one day to remove the moisture content.
Samples will be ground into a fine powder to increase the surface contact with the solvent extract
and stored in airtight containers for further use. The extracted oil was then filtered and stored for
further use.

 3.4 SOXHLET EXTRACTION 

 Soxhlet extraction is a specific type of liquid-liquid extraction. It is a method used to extract
lipids from a sample. The Soxhlet apparatus is a large glass tube connected to a heating element
and a condenser. The sample is placed into a thimble, which is placed into the Soxhlet apparatus.
The sample is then heated, and the liquid extract is collected in a graduated cylinder. The Soxhlet
method removes lipids from a sample, such as oils and fats, without destroying the sample. This
method helps extract lipids from various samples, such as food and soil samples. The solvent
used in this extraction process typically combines hexane and ethanol.

PROCEDURE 
10g of crushed pumpkin seed will be placed inside a thimble made from thick filter paper, which
will be loaded into the main chamber of the Soxhlet extractor. The Soxhlet extractor will be
placed onto a flask containing the extraction solvent. The Soxhlet will be equipped with a
condenser. The solvent will be heated to reflux. The solvent vapour travels up a distillation arm
and floods into the chamber, housing the thimble of the solid. The condenser ensures that any
solvent vapour cools and drips back into the chamber housing the solid material. The chamber
containing the solid material fills with warm solvent. Some of the desired compounds will then
dissolve in the warm solvent. When the Soxhlet chamber is almost complete, the chamber will be
automatically emptied by a siphon side arm, with the solvent running back to the distillation
flask. This cycle may be repeated many times over hours or days. During each cycle, a portion of
the non-volatile compound dissolves in the solvent. After many cycles, the desired compound is
concentrated in the distillation flask. The advantage of this system is that instead of many
portions of warm solvent being passed through the sample, just one batch of solvent is recycled.
After extraction, the solvent will be removed, typically using a rotary evaporator, yielding the

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extracted compound. The non-soluble portion of the extracted solid remains in the thimble and
will be discarded.
Apparatus and reagent 
Mortar and pestle 
Hexane (solvent) 
Heating mantle 
Measuring cylinder 
Weighing balance 
Soxhlet extractor 
  

3.5 COLD PRESSING 

Cold pressing is a method of extracting lipids from plants and other sources. This method uses
mechanical pressure to break down the source material's cellular structures, releasing the lipid
contained. The mechanical pressure is typically applied using a hydraulic press, which can be
adjusted to produce different pressure levels depending on the source material and desired
outcome.
PROCEDURE 
50g of the crushed will be placed in a food processor and grind until they are a fine paste. Heat a
heavy-bottomed pan over medium heat, and ground pumpkin seeds will be added. The pumpkin
Seed paste will be cooked until lightly browned and stirred occasionally. The paste will be cool
slightly and then transferred to a clean cloth; it will then be Squeeze extract oil from the pumpkin
seed paste. The cold-pressed pumpkin seed oil will be stored in an airtight container in a cool,
dark place.
Apparatus and reagent 
Clean tap water 
Mortar and pestle 
Heating pan 
Volumetric flask

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 3.5 ANALYSIS OF THE OIL EXTRACTED 

3.5.1 Determination of Oil Yield

The oil yield depends on the processing parameters. The extraction yield was determined from
the relationship between the oil extracted and the raw material used.

procedure
The sample weight will be measured before pumpkin seed oil extraction and denoted by M1.
After the extraction process, the sample will be weighed, and the oil is complete and will be
dried in an oven denoted by M2.
The yield of oil (%) is calculated
(M1-M2)/M1×100
Where m1 denotes the sample's weight before extraction, and M2 denotes the weight after
extraction, which dried in the oven.

3.6.2 IODINE VALUE 

Procedure 
All the prepared reagent solutions and the requirements on the table will be arranged.
2ml of the oil sample will be pipetted and dissolved in Chloroform to a round bottom flask
labelled "TEST."
10ml of Iodine Monochloride reagent will be added into the flask. The contents in the flask will
be mixed thoroughly.
Then the flask will be allowed to stand for half an hour of incubation in the dark.
The BLANK will be set in another iodination flask by adding 5ml Chloroform.
10ml of Iodine Monochloride reagent will be to the BLANK, thoroughly mixing the contents in
the flask.
The BLANK will be placed in the dark for 30 minutes.
The TEST will be removed from incubation after 30 minutes, and 10ml of potassium iodide
solution will be added to the flask.
The stopper and the sides of the flask will be rinsed using 50 ml of distilled water.

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The “TEST” will be titrated against standardized sodium thiosulphate solution until a pale straw
colour is observed. 
1ml starch indicator will be in the flask's contents, and purple will be observed.
The titration will proceed until the colour of the solution in the flask turns colourless.
The disappearance of the blue colour will be recorded as the end point of the titration. 
Similarly, the procedure will be repeated for the flask labelled 'Blank.'
The endpoint values will be of the BLANK  
Calculate the iodine number will be done using the equation by
The volume of Sodium thiosulphate used = [Blank- Test] ml 
 Equivalent Weight of Iodine = 127g/mol 
IV=(V1-V2) *N*12.69)/W 
Where, 
V1 and V2 are sodium thiosulphate volume 
N is the normality sodium thiosulphate 
W is the weight of the oil taken 
12.69 is the conversion factor from mEQ sodium thiosulphate to grams of iodine 
Normality of sodium thiosulphate (Na2S203) = 0.1 

3.6.3 ACID VALUE 

PROCEDURE 

The oil will be mixed thoroughly before weighing. 2ml of oil will be Weighed accurately and
placed in a 200 ml conical flask. The weight of the oil will be taken for the test, and the strength
of the alkali used for the titration will be such that the volume of alkali required for the titration
will not exceed 10 ml. 50 ml of fresh hot ethyl alcohol and about 1 ml of phenolphthalein
indicator solution will be added. The mixture will be boiled for about five minutes and titrated as
hot as possible with standard aqueous alkali solution, shaking vigorously during titration.
CALCULATION 
Acid value = (56.1 × N × V)/W 
  
Where, 

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V = volume in ml of standard KOH/NaOH solution used, 
N = normality of standard KOH/NaOH solution, and 
W = weight in g of the material taken for the test 
  Yield
  

DATA ANALYSIS

The data analysis will involve comparing the acid value and iodine value of the oil obtained from
both extraction methods. The acid value measures the amount of free fatty acids in the oil and is
calculated by titrating the sample with a standard potassium hydroxide (KOH) solution. The
iodine value measures the degree of unsaturation of the oil and is calculated by titrating the
sample with a standard sodium thiosulphate solution. The results of each titration will be
compared to national and international standards, such as those set by the Food and Agriculture
Organization of the United Nations/World Health Organization (FAO/WHO) recommendation
(2003), the permissible level of acid value for all edible oils should be below 0.6 mgKOH/g
(measured in potassium hydroxide per gram), as stated in the AOCS Official Method Cd 8-53
(FAO/WHO, 2003). Additionally, this is also supported by the ISI Handbook of Food Analysis
(Part III, 1984) and method of sampling and test for oils and fats, as well as the ISO 6601 (1996)
for the determination of Acid value and acidity while the iodine value should be between 120-
140 g/100g oil for edible oils. The yield will be calculated using the following:
The yield of oil (%) is calculated
(M1-M2)/M1×100

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 Where m1 denotes the sample's weight before extraction, and M2 denotes the weight after
extraction, which will be dried in the oven.

Appendix

Budget 

table 1.1 1. laboratory utilities, resources, and cost estimating

ITEMS    COST PER UNIT KSH TOTAL COST KSH

Typing(20pages)    10    200
Pumpkin seeds 1kg    285    570
hexane printing(20pages)    10    200
Laboratory utilities    NIL   
TOTAL COST KSH    305 970

Work plan 

table 1.1 2. activity work and period to be done

Months February March April May

Activity

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Proposal title
submission
Project proposal
writing
Presentation of
project proposal
Data collection
Research project
writing
Project presentation

  

4. References 

Hussain, M. A., Zia-Ul-Haq, M., Ali, A., Asghar, A., Rehman, A., & Ullah, S. (2020). Extraction
and characterization of oil from the pumpkin seed. International Journal of Food Properties,
23(2), 547–555.Aregheore, E.M. (2014). Edible Oil Processing. CABI. 
Huang, X., Li, G., Li, J., Li, C., Li, Y., Li, F., & Li, X. (2019). Lipid Extraction from Plant
Tissues Using Acetone and Ethanol: A Comparison. Plants, 8(2), 34.
https://doi.org/10.3390/plants8020034 
Liu, Z., & Chen, C. (2011). Extraction of pumpkin seed oil using supercritical carbon dioxide
and conventional solvent. Journal of Food Engineering, 106(2), 216-222.
doi:10.1016/j.jfoodeng.2011.02.013  
  Pumpkin seed oil: A source of health and nutrition. Journal of Food Science and Technology,
54(2), 459–465. Hajebrahimi, S., Asghari, G. R., & Omidian, H. (2017). Physicochemical
properties of pumpkin seed oil as affected by extraction methods. International Journal of Food
Properties, 20(11), 2328–2337. Kaur, M., Sharma, B., & Kaur, G. (2018). Pumpkin seed oil: A
comprehensive review of its therapeutic potential. Clinical Phytoscience, 4(1), 6. Sarker, M. A.,
Uddin, M. S., & Chowdhury, M. R. (2020). Nutritional and health benefits of pumpkin seed oil:
A review. Asia Pacific Journal of Clinical Nutrition, 29(3), 547–556.

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Ereifej, K. S., Brenna, J. T., Carroll, K. K., & Erdman, J. W., Jr. (2017). Composition and
antioxidant activity of pumpkin seed oil. Journal of Food Science, 82(7), 1545-1551.
https://doi.org/10.1111/1750-3841.13687 
Bhardwaj, S., Sharma, R., & Dixit, S. (2017). Potential Health Benefits of Pumpkin Seed Oil: A
Review. Journal of Nutrition & Food Sciences, 7(3), 563–567. https://doi.org/10.4172/2155-
9600.1000563 
 Durante M., Lenucci M. S. and Mita G.2014. Supercritical carbon dioxide extraction of
carotenoids from pumpkin (Cucurbita spp.): a review. Int. J. Mol. Sci. 15:6725-6740. DOI:
doi.org/10.3390/ijms15046725

  
 
 
 

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