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Thin Layer Chromatography (tlc) for Vegetables

Autor:   •  April 14, 2018  •  Lab Report  •  2,077 Words (9 Pages)  •  95 Views

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FACULTY OF BIORESOURSE AND FOOD INDUSTRY

2014/2015 SEMESTER II

FIS 3122

FOOD CHEMICAL ANALYSIS

Experiment 4: Thin Layer Chromatography of Vegetables

Name: Siti Nadhira bt Mohamad Basri

ID Number: 034505  

Group: 1

Course: Bachelor of Food Technology (Hons)

Instructors:

1. Prof. Madya Dr. Kamarul ‘Ain Mustafa

2. Dr. Nurul Zaizuliana Rois Anwar

3. Pn. Tengku Farizan Izzi Che Ku Jusoh

4. En. Roslan Arshad  

Objectives  

1. To conduct Thin Layer Chromatography (TLC) tests on spinach and carrot.  

2. To determine the retardation factor, Rf value for spinach and carrot.

3. To understand the principle of Thin Layer Chromatography (TLC).  

Procedures

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Results    

Distance solvent moved from the spotting line (origin): 5cm

Table 1: Spinach

Trial

Colour of spot

Distance moved

(cm)

Rf value

1

Light yellow

1.0

0.2

Yellowish-green

1.5

0.3

Yellow

1.75

0.35

Green

2.0

0.4

Light green

2.2

0.44

2

Light yellow

0.7

0.14

Light yellow

1.4

0.28

Yellowish-green

1.75

0.35

Yellowish-green

1.9

0.38

Green

2

0.4

Light green

2.2

0.44

Table 2: Carrot  

Trial

Colour of spot

Distance moved

(cm)

Rf value

1

Yellowish

1.7

0.34

Light orange

2.6

0.52

-

-

-

2

Yellowish

0.7

0.14

Light orange

1.7

0.34

Orange

2.7

0.54

Calculations:  

Rf value = Distance from origin to component spot

                Distance from origin to solvent front  

Table 3: Rf value of each spot for spinach

Trial

Colour of spot

Distance moved

(cm)

Calculations  

(cm)

Rf value

1

Light yellow

1.0

Rf = 1.0/5

0.2

Yellowish-green

1.5

Rf = 1.5/5

0.3

Yellow

1.75

Rf = 1.75/5

0.35

Green

2.0

Rf = 2.0/5

0.4

Light green

2.2

Rf = 2.2/5

0.44

2

Light yellow

0.7

Rf = 0.7/5

0.14

Light yellow

1.4

Rf = 1.4/5

0.28

Yellowish-green

1.75

Rf = 1.75/5

0.35

Yellowish-green

1.9

Rf = 1.9/5

0.38

Green

2.0

Rf = 2.0/5

0.4

Light green

2.2

Rf = 2.2/5

0.44

Table 4: Rf value of each spot for carrot

Trial

Colour of spot

Distance moved (cm)

Calculation  

(cm)

Rf value

1

Yellowish

1.7

Rf = 1.7/5

0.34

Light orange

2.6

Rf = 2.6/5

0.52

-

-

-

-

2

Yellowish

0.7

Rf = 0.7/5

0.14

Light orange

1.7

Rf = 1.7/5

0.34

Orange

2.7

Rf = 2.7/5

0.54

Discussion

Chromatography is used to separate mixtures of substances into their components. All forms of chromatography work on the same principle. They all have a stationary phase and a mobile phase. The mobile phase flows through the stationary phase and carries the components of the mixture with it. Different components travel at different rates. Thin layer chromatography is done using a thin, uniform layer of silica gel or alumina coated onto a piece of glass, metal or rigid plastic. In this experiment, the silica gel is the stationary phase and the mobile phase is 70:30 hexane-acetone solvent. The samples used are spinach and carrot.  

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