Biochemistry Lab Essay, Research Paper
I. Introduction
In this experiment, lipoids from land Myristica fragranss are extracted utilizing a combination of dissolvers and place the lipoids through chromatography. The intent of utilizing solvent combinations is to elute the lipoids based on their mutual opposition to binding of the silicon oxide gel. The chromatography is performed on a silicon oxide gel home base and the usage of I to visualise the lipoids. By ciphering the Rf values for each compound and comparing them to the known lipoids, we are able to separate the lipoids within the grounded Myristica fragrans.
II. Procedures
Add 5 g crushed nutmeg and 50 mL hexane-isopropanol into a flask and warm for 15 proceedingss.
Remove the excess dissolver on a steam bath under a goon while blushing the flask with N2 gas, go forthing the petroleum infusion. Weigh infusion.
Prepare silica gel column. Add 6 g of silicon oxide gel in 20 milliliter of hexane to do a slurry. Block column with little piece of glass wool, add 5 milliliter of hexane and so add the silicon oxide slurry up to the 10 centimeter grade.
Prepare.05 to.075 g of petroleum lipid by fade outing it in hexane. Add to the silicon oxide gel slurry in the column.
Get down roll uping samples with the pure hexane. Keep adding hexane so that the silicon oxide gel column does non run dry. Roll up one 20 milliliter sample. Repeat with 90:10 hexane and roll up 4 20-mL bottles. Repeat with 80:20 hexane and roll up 2 20-mL samples.
Analyze each fraction by descrying 10 times with capillary tubings on a TLC home base, which is exposed to iodine vapour for 15 proceedingss.
III. Consequences
Lipids Distance Traveled Rf & # 8211 ; Correct Results
Rf = Distance Solute Traveled / Distance Solvent Traveled ( millimeter )
Hexane- 0
Hexane 90:10- 123/140 = .879
Hexane 90:10- 39/141= .277
Hexane 90:10- 38/141= .270
Hexane 90:10- 0
Hexane 80:20- 20/139= .863
Hexane 80:20- 19/139= .137
Triolein- 107/137= .78
Oleic Acid- 44/143= .309
Cholesteryl Linolet- 141/142= .99
Crude Lipid- ( 10 musca volitanss )
0, .035, .063, .092, .155, .204, .303, .45, .669, .99
Lipids Distance Traveled Rf & # 8211 ; Incorrect Results
Hexane- 0
Hexane 90:10 ( All 4 samples ) – 0
Hexane 80:20 ( Both Samples ) – 0
Triolene- 85/149= .570
Oleic Acid- 55/149= .369
Cholesteryl Linolet- 150/150= 1
Crude Lipid- 15 musca volitanss
IV. Discussion
In this experiment, extracted lipoids are compared to three known lipoids listed supra. The consequences obtained for our group were classified as incorrect due to unknown experimental mistake. Therefore, consequences from another group were attained for treatment intents. In looking at the consequences it can non be clearly identified what the unknown lipoid is. The Rf values from the three criterion Ar
e triolene: .78, oleic acid: .309, and cholesteryl linolet: .99. The petroleum lipoid produced 10 different musca volitanss. Topographic point 9: .669 is reasonably near to the standard triolene. On the petroleum consequences, topographic point 10: .99 is near to the standard cholesteryl linolet. The remainder of the hexane 90:10 and 80:20 samples did non fit up to the criterions. The hexane was supposed to bring forth a 0 Rf values since it is nonionic and would non be expected to go on the TLC home base. Theoretically, the hexane infusion, a really nonionic compound, should hold traveled further than the 90:10 and 80:20. In looking at the consequences this is precisely what happened since the concluding value of the lipoid was.99. The Rf value of the hexane infusion should be nigh one as those of cholesteryl linolet or oleic acid because the silicon oxide gel is polar. The 80:20 infusion is the most polar infusion every bit far as this experiment is concerned and it should hold the lowest Rf value since it will adhere to the silicon oxide gel home base and will non go with the solvent forepart every bit readily as nonionic molecules. The 80:20 should hold Rf values near the oleic acid, since both are polar. As for the 90:10, the Rf value does non propose that it is any of the standard lipoids, nevertheless, during the ocular review, its location appeared to be in the same place as triolein. The Rf value of triolein and that of the 90:10 infusion are the nearest to each other with a fringy sum of mistake. The mistake may be because improper measurings were made because the solute has migrated horizontally from its original topographic point, giving it a smaller value so would be expected.
In comparing these consequences with our group & # 8217 ; s consequences, one can see many discreprencies. First of wholly, the hexane samples in all three concentrations did non bring forth any consequences. The oleic acid and cholesteryl linolet had right values fiting up to the other group & # 8217 ; s consequences. But the triolene had a value of.57 and the triolene sample from the other group was.78, demoing a difference. The major ground that the consequences may hold been different is experimental mistake. While roll uping samples, the column went dry at one juncture. This may hold produced samples with wrong sums of lipoid in it. The silicon oxide gel column has to be kept running with fluids at all times. Otherwise, the consequences will be wrong, as our consequences were.
V. Conclusion
The experiment served its intent by showing the features of lipoids. The utile techniques of surface assimilation chromatography on a silicon oxide gel column and analysis by thin bed chromatography proved to be utile techniques. In my sentiment these techniques should be learned by anyone that wants to travel in a biology/biochemistry calling. This experiment besides proved how of import experimental technique and process is. A small error or experimental mistake, such as the column drying, can bring forth extremely wrong consequences. Overall, the experiment was really gratifying and fatty acids can be concluded as a nonionic with little mutual opposition substance.
