Thin Layer Chromatography (tlc) for Vegetables
Essay by Nadhira Kim • April 14, 2018 • Lab Report • 2,077 Words (9 Pages) • 1,431 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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