FACULTY OF SCIENCE

BACHELOR OF SCIENCE (HONOURS) DIETETICS

UDDN 1304 NUTRITIONAL BIOCHEMISTRY

EXPERIMENT TITLE

(Determination of Crude Lipid of Vegetable Seeds)

STUDENT NAME STUDENT NO. COURSE YEAR/SEM SIGNATURE

TAN SEE THEAN 2000151 DT 1/1

TEOH HWEI FANG 2000150 DT 1/1

SIOW YEN YEN 1907048 DT 1/2

ASLINDA JIE BT 1906353 DT 1/1

ISMAIL

UNG KAH YENG 1806958 DT 1/1

LECTURER: DR. TAN CHIN XUAN

PRACTICAL GROUP: P3 (GROUP 1)

DATE OF SUBMISSION: 11 March 2020

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 1.0 Objectives

● To determine the crude lipid of vegetable seed using the Soxtherm method.
● To familiarize with the Soxhlet method.

2.0 Principles

Oil and fat from the solid seed powders are extracted by repeatedly washing the sample with an organic
solvent under reflux, where in this experiment, petroleum ether was the organic solvent used. This method
is known as percolation. A continuous evaporation and condensation of the solvent is known as reflux.
Reflux is often used because oil is heavy and can form sediments throughout the process. By refluxing the
solvent, the solvent is vaporized in pure form, therefore, decreasing the impurities in the extractions.

3.0 Procedures

A. Extraction Beakers Preparation

1. The extraction beakers were preheated together with the boiling stones (3pcs) at 105
degree Celsius for 1 hour in the drying oven.
2. The extraction beakers were cooled down in desiccators.
3. The extraction beakers were weighed and recorded as M1 value.

B. Sample Preparation

1. The sample was made sure it was homogenized. Sample with high fat=1-3g, and sample
weight was 6g.
2. The sample on the filter paper was weighed and recorded as M0 value.
3. The sample with filter paper was wrapped and the bottom of a thimble was inserted.
4. A piece of cotton wool was inserted on the top of the sample.
5. The thimble was put into a thimble holder and then they were inserted into the extraction
beaker.
6. The amount of solvent was poured in certain into the extraction beakers in fume hood.
7. The sample was extracted in boiling solvent during the first step.
8. The level of the solvent was reduced until there was a clear separation between the
solvent and the extraction thimble. The distillate was collected in the recovery tank.
9. The material was extracted from the sample over the boiling solvent by the refluxed,
condensed solvent.
10. The bulk of the solvent was distilled off into the storage tank for later recovery.
11. The extraction beakers were lifted from the hotplate automatically. The evaporation
process continued slightly, without adverse effects on the sample. Coolant and heater
were atomically switched off when extraction was finished after the fifth stage.

C. After Extraction:

1. The extraction beakers were taken out and put into the insert rack with aid of a clamp.
Extraction beakers should not be handled with bare hands.
2. The thimble holder together with the thimble was removed from extraction beakers in the
fume hood.

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 3. The extraction beakers were heated at 105 degree Celsius for 1 hour in the dying oven.
4. The extraction beakers were cooled in desiccators for about 1 hour.
5. The extraction beakers were weighed and recorded as M2 value.

4.0 Results and Calculations

Results

Table 1: Crude lipid content of vegetable seeds

Type of vegetable seeds Bitter gourd Cucumber Snack gourd

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 Sample weight (g) R1: 6.10 R1: 6.00 R1: 6.02

R2: 6.12 R2: 6.15 R2: 6.00

Empty extraction beaker R1: 127.63 R1: 126.88 R1: 126.08

R2: 125.66 R2: 126.50 R2: 126.36

Extraction beaker with R1: 145.27 R1: 144.19 R1: 142.95

lipid (g)
R2: 142.75 R2: 143.92 R2: 142.75

Lipid (%) R1: 289.18 R1: 288.50 R1: 280.23

R2: 279.25 R2: 283.25 R2: 273.17

Average ± Standard 284.22±7.02 285.88±3.71 276.70±4.99

deviation

Remarks: R1: Reading 1 and R2: Reading 2

*When measuring the weight of extraction beaker with lipid, we did not take out the timber holder and
timber, so there is an error occurred in the percentage of lipid, average and standard deviation of result.

5.0 Discussion

Lipids are organic compounds insoluble in water and soluble in organic solvent such as alcohol,
ether and chloroform. The major lipids are fatty acids, triglycerides, phospholipids and sterols. The most
abundant lipids in the foods are fatty acids, it consists of different forms of fatty acids including saturated
fatty acids, unsaturated fatty acids and trans fatty acids (McNight, 2017). The fatty acids can be obtained
either from animal or plant sources.

Saturated fatty acids (SFAs) are fat molecules containing carbon-carbon single bonds, it exists as
solid form in room temperature. Animal sources contribute the majority of dietary saturated fatty acids
(SFAs). Examples of food with saturated fatty acids are fatty beef, lamb, poultry with skin, butter, cheese
and other dairy products (Moll, 2019). Secondly, fatty acids that contain one or more double bonds are

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 unsaturated fatty acids and it exists as liquid form at room temperature. The unsaturated fats can be
further classified into two which are monounsaturated fats and polyunsaturated fats. Monounsaturated
fatty acids (MUFAs) are fatty acids with only one double bond and mostly found in plant based foods
which are olive oil, almonds, cashews, peanuts and avocados (Robertson, 2017). Polyunsaturated fatty
acids (PUFAs) are fatty acids that contain two or more double bonds and it can be obtained either from
animal and plant based foods. Animal based foods such as salmon, herring, sardines and mackerel provide
a high amount of PUFAs that are rich in omega-3. Whereas, plant source oil also provides PUFAs that are
rich in omega-6 fatty acids like safflower seed oil, grapeseed oil, flaxseed oil and sunflower seed oil
(Walle, 2018). Apart from that, trans fatty acids can be found naturally in dairy and beef products
(McGuire and Beerman, 2013). unsaturated fats found primarily in partially hydrogenated vegetable oils
and foods containing these oils and in ruminant (animal) fats.

Lipids are one of the important macronutrients because they provide a source of energy during
critical time such as fasting and starvation. Fats can be stored around delicate internal organs in order to
protect the organs, for example, the kidney. Fats also form the living cell membrane. Most cell
membranes feature phospholipid bilayers. In addition, the lipid component allows the cell membrane to
remain hydrophobic, which prevents certain molecules from moving through. Fats act as insulation that
form layers of fat under the skin insulate the body when temperatures drop by generating heat through
metabolism (Bailey, 2020). It also plays a role in making hormones such as prostaglandin which regulate
blood pressure (McGuire and Beerman, 2013).

Recommended daily allowance is intended to help people obtain adequate nutrition for health and
wellness. The recommended intake for lipids is less than or equal to 30% of calories from total fat, less
than or equal to 10% of calories from saturated fat, and no more than 300 mg of cholesterol. The
Acceptable Macronutrient Distribution Ranges (AMDR) recommend that healthy adults consume 20 to
35% of their energy from lipid. If a person requires 2,000 kcal per day, he or she should obtain about 400
to 700 kcal from the dietary lipids. The energy content for 1 gm of fats produced 9 kcal of energy. Hence,
the amount of dietary intake of fats needed for most adults are 44 to 78 g (McGuire and Beerman, 2013).
So the exact gram requirement for the intake of total dietary fats depends on the average number of
calories in the diet (Vercelletto, 2019).

There are few consequences of excess lipid consumption like obesity and cardiovascular disease.
If too much fat is consumed, it will have the potential for weight gain. The reason that fat is easy to gain
in weight is because it has higher calories compared to other nutrients which is 9 calories per gram. When
humans consume excess calories per day it can lead to weight gain and obesity (Tremblay, 2018).
Besides, truncal obesity is a risk factor for heart attack. Thus, there is a risk of cardiovascular disease for
overeating fats. Saturated and trans fats increase the level of harmful cholesterol in our bloodstream.
However, there are some consequences of inadequate lipid consumption. Fats played important roles in
absorption of fat soluble vitamins like vitamins A, D, E and K, so inadequate lipid consumption will
cause deficiencies in fat-soluble vitamins. Furthermore, essential fatty acids are required to build and
maintain our cell membranes, which is important for our skin, hair, eyes, heart and so on. Therefore, there
are some signs that our body lacks fats. It was dry and scaly skin, dry eyes, feeling constantly cold which
because fats act as insulation that form layers of fat under the skin insulate the body when temperatures
drop by generating heat through metabolism. Nevertheless, there are also signs of dry hair, hormonal
problems, always feeling hungry, and constant fatigue because fats are a source of energy (Steen. 2017).

There is an error occurred in the percentage of lipid and so that the average and standard
deviation results, because the timber holder and timber were not taken out from the extraction beaker with
lipid before weighing. However, the comparison based on the above results showed that cucumber has the
highest fats among three of the samples. Cucumbers have an average 285.88% of lipid, while bitter gourd
and snake gourd are 284.33% and 276.70% respectively.

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 References

Bailey, R. (2020). Fats, Steroids, and Other Examples of Lipids. [online] Available at:
<https://www.thoughtco.com/lipids-373560> [28 February 2020].

McGuire, M. and Beerman, K. A., 2013. Nutritional Sciences. Third edition. United States: Wadsworth
Cengage Learning.

Moll, J. (2019). The Difference Between Saturated and Unsaturated Fats. [online] Available at:
<https://www.verywellhealth.com/difference-between-saturated-fats-and-unsaturated-fats-697517> [29
February 2020].

Robertson, R. (2017). What Are the Benefits of Monounsaturated Fats? [online] Available at:
<https://www.healthline.com/nutrition/monounsaturated-fats> [1 March 2020].

Steen, J. (2017). 9 Signs You’re Not Eating Enough Fat. [online] Available at :
<https://www.huffingtonpost.com.au/2017/11/19/9-signs-youre-not-eating-enough-fat_a_23279029/> [6
March 2020]

Tremblay, S. (2018). Health Risks Associated With Excessive Intake of Fats and Carbs. [online] Available
at: <https://healthyeating.sfgate.com/health-risks-associated-excessive-intake-fats-carbs-5365.html> [6
March 2020]

Vercelletto, C. (2019). Here’s How Much Fats You Should Actually Be Eating on the Daily. [online]
Available at: <https://www.livestrong.com/article/370685-how-many-fat-grams-per-day/> [1 March
2020].

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 Walle, G. V. D. (2018). Polyunsaturated Fats: Know the Facts About These Healthy Fats. [online]
Available at: <https://www.healthline.com/nutrition/polyunsaturated-fat#what-are-they> [2 March 2020].

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