SBL 1023

Technique in Biology And

Biochemistry Laboratory

Lab 4: lipid analysis

NOR AIYNA SYERA BT MOHD ASRI

STUDENT`S NAME
( E20161015981)

GROUP B

DATE/DAY/PLACE 21/11/2017 /TUESDAY/ B2-L3-MP 14

LECTURE`S NAME Professor Madya Dr. Shakinaz Binti Desa

Tittle

- Lipid analysis

Objective

- To determine the amount of protein in lipid sample.

- To find the percent of lipid extraction in the food sample.
- To see the reaction of lipid with Sudan
- To test the solubility of the lipid

Introduction

Lipids are one of the major constituents of foods, and are important in our diet for a
number of reasons. They are a major source of energy and provide essential lipid nutrients.
Nevertheless, over-consumption of certain lipid components can be detrimental to our health,
e.g. cholesterol and saturated fats. In many foods the lipid component plays a major role in
determining the overall physical characteristics, such as flavor, texture, mouthfeel and
appearance. For this reason, it is difficult to develop low-fat alternatives of many foods,
because once the fat is removed some of the most important physical characteristics are lost.
Finally, many fats are prone to lipid oxidation, which leads to the formation of off-flavors
and potentially harmful products. Some of the most important properties of concern to the
food analyst are:

 Total lipid concentration

 Type of lipids present
 Physicochemical properties of lipids, e.g., crystallization, melting point, smoke point,
rheology, density and colour.
 Structural organization of lipids within a food

Lipids are usually defined as those components that are soluble in organic solvents
(such as ether, hexane or chloroform), but are insoluble in water. This group of substances
includes triacylglycercols, diacylglycercols, monoacylglycercols, free fatty acids,
phospholipids, sterols, caretonoids and vitamins A and D. The lipid fraction of a fatty food
therefore contains a complex mixture of different types of molecule. Even so,
triacylglycercols are the major component of most foods, typically making up more than 95
to 99% of the total lipids present. Triacylglycerols are esters of three fatty acids and a
glycerol molecule. The fatty acids normally found in foods vary in chain length, degree of
unsaturation and position on the glycerol molecule. Consequently, the triacylglycerol fraction
itself consists of a complex mixture of different types of molecules. Each type of fat has a
different profile of lipids present which determines the precise nature of its nutritional and
physiochemical properties. The terms fat, oil and lipid are often used interchangeably by food
scientists. Although sometimes the term fat is used to describe those lipids that are solid at
the specified temperature, whereas the term oil is used to describe those lipids that are liquid
at the specified temperature.

Sudan stains are synthetic organic compound that are used to dye food and other
biological sample, especially that contain lipid. Sudan has height affinity to lipids, so they are
often used as marker to test the presence of lipid in a sample. Like lipids, Sudan stain is not
soluble in water: it is however, soluble in lipids. If lipids are present, the Sudan stain will
stain them reddish-orange (positive test).

Equipment and materials

- Lipid samples (cooking oils, margarine, vegetable shortening, lecithin)

- Food samples (choki-choki)
- Petroleum ether
- Deionised water
- Test tubes and test tube rack
- Measuring cylinder
- Dropper and spatula
- Erlenmeyer flask.
- Beaker
- Weighing scale
- Hot plate
Methodology

(A) Extraction of lipids from foods.

1. Food sample such as choki-choki flavor peanut and butter was weighed about 2 gram.
2. Food sample was crushed into small pieces used mortar and pestle.
3. Empty beaker with a glass rod inside the beaker was weighed and the data was
recorded in Table 1.0.
4. The crushed food sample was placed in the beaker.
5. The beaker (with a glass rod) that contained the crushed food was weighed and data
was recorded in Table 1.0.
6. 10 ml of petroleum ether was added to the flask contained the crushed food. Then, the
crushed food with petroleum ether was stirred with the glass rod for 5 minutes to get
the lipids to dissolve in the petroleum ether.
7. The petroleum ether was carefully decant from the beaker.
8. The beaker containing the remaining solid food was placed on a hot plate to evaporate
all of the petroleum ether.
9. After all of petroleum ether was evaporated, the smell characteristics of the petroleum
ether was checked until no longer strong smell, and then, allowed the beaker to cool.
10. The beaker and remaining food was weighed again (with the glass rod) after the
beaker has cooled to room temperature. Then, the weigh was recorded in Table 1.0
11. By using Table 2, the mass of lipid extracted from the food sample was calculated and
the weight percent of lipid in that food was determined.
Result

Weight of Weighed of Weight of Weight lost

empty beaker with Weighed of beaker with from food
% lipid
Food sample beaker crushed raw crushed raw dried food (weight of
extraction
(with glass food (with food, g (with glass lipid
rod) , g glass rod) , g rod), g extracted) , g

66.00 g - 66.00 g –
Choki-choki
63.95 g 66.0 g 63.95 g = 65.40 g 65.40 g = 29 %
(chocolate)
2.05 g 0.60 g

Choki-choki 67.99 g – 67.99 g –

(chocolate + 66.05 g 67.99 g 66.05 g = 67.43 g 67.56 g = 29 %
peanut) 1.94 g 0.56 g

Choki-choki 61.95 g – 61.95 g –

(chocolate + 59.80 g 61.95 g 59.80 g = 61.22 g 61.22 g = 29 %
vanilla) 2.15 g 0.73 g

Table 1: Extraction of Lipid

Choki-choki
(chocolate)

Choki-choki
(chocolate + peanut)

Choki-choki
(chocolate + vanilla)

Table 2: Image of the sample

 Weighed new beaker Weight of beaker
Food sample without decant with dried decant % lipid
petroleum ether , g petroleum ether, g

Choki-choki
65.20 g 45.17 g
(chocolate) 31 %

Choki-choki
65.25 g 50.15 g
(chocolate + peanut) 23 %

Choki-choki
65.70 g 32.67 g
(chocolate + vanilla) 50 %

Table 3: % lipid

Calculation

weight of lipid extracted

% Lipid extraction = × 100%
weight of crushed food sample

weight petroleum ether before heated −

weight of petroleum ether after heated (dry)
% Lipid = × 100%
weight of petroleum ether before heated

1. Choki-choki (chocolate)

0.60 𝑔
% 𝐿𝑖𝑝𝑖𝑑 𝑒𝑥𝑡𝑟𝑎𝑐𝑡𝑖𝑜𝑛 = × 100%
2.05 𝑔

= 29 %

65.20 𝑔 − 45.17 𝑔
% 𝐿𝑖𝑝𝑖𝑑 = × 100%
65.20 𝑔

= 31 %
 2. Choki-choki ( chocolate + peanut)

0.56 𝑔
% 𝐿𝑖𝑝𝑖𝑑 𝑒𝑥𝑡𝑟𝑎𝑐𝑡𝑖𝑜𝑛 = × 100%
1.94 𝑔

= 29 %

65.25 𝑔−50.15 𝑔
% 𝐿𝑖𝑝𝑖𝑑 = × 100%
65.25 𝑔

= 23 %

3. Choki-choki (chocolate + vanilla)

0.73 𝑔
% 𝐿𝑖𝑝𝑖𝑑 𝑒𝑥𝑡𝑟𝑎𝑐𝑡𝑖𝑜𝑛 = × 100%
2.15𝑔

= 29 %

65.70 𝑔 − 32.67 𝑔
% 𝐿𝑖𝑝𝑖𝑑 = × 100%
65.70 𝑔

= 50 %
Discussion

From the table above, we can see that choki-choki with different type of flavour has
the same percentage of lipids extracted from it. The percentage of extraction lipid of the
sample is shown in Table 2. The percentage of lipid extraction between chocolate, chocolate
+ peanut and chocolate + vanilla are same, 29%. Then, from the result, percentage of lipid of
the sample is shown in Table 3. From the table, the percentage of lipid in chocolate is 31%,
chocolate + peanut is 23% and chocolate + vanilla is 50%. So, we can conclude that Sample
C form the higher percentage of lipid. This is because Sample C has combination of two type
of flavour. It is chocolate and vanilla. But from my opinion, Sample B will get a higher
percentage of lipid. This is because, in the Sample B contains a peanut. Peanut is
monosaccharides and it contains higher of lipids. So, we can conclude that the peanut in the
sample is just as a flavor that does not affect any lipid content in it. It just a flavour that not
contain any real of peanut.

There are some tips to prevent the error during the experiment. First, we must ensure
that we used a dry beaker because any water or other chemical present on the beaker may
affect the experiment and consequently the result. Then, we must to stir the beaker by using a
glass rod after placed the petroleum ether. It is important to ensure any lipid in the sample has
been dissolve.

Conclusion

From the experiment, according to the result, we can conclude that the lipid extracted from
the chocolate consists of saturated fatty acid and the chocolate contains less of lipid.

References

1. Lab 4: lipid analysis. (2017). SBL 1023 LAB MANUAL. Faculty science and
mathematic.
2. McNaught, A. D., & Wilkinson, A. (1997). Compendium of Chemical Terminology
(the "Gold Book"). Oxford: Blackwell Scientific Publications.