Hi, I need to perform experiment for separation of lipids sample by TLC (for learning purpose to students). Samples are: soybean oil, mustard oil, butter. I need to choose suitable solvent system to run TLC????
I found these two:
1. petroleum ether: diethyl ether: glacial acetic acid in the ratio of 80:20:1
2. chloroform: acetone: methanol: acetic acid: water (100:40:20:20:10)
Which one should I go for?
According to my experience in this field
In each client separated by TLC, several types of mobile phase must be used
Then depend on the mobile phase that gave the highest number of compounds
The best separation between vehicles without interference
You can also use two-dimensional
First, use a single number moving phase
Then use another phase after turning the plate on its side
In this way. Use one spot of the extract on one side of TLC
After the completion of the migration, flip on the side where you are traveling and place in the second mobile phase.
This method ensures a clear contrast of compounds without interference
Hi Nabin
When analysing lipid clases, I use hexane/diethyl ether/acetic acid (80:20:2, by vol), as neutral solvent, to separate free fatty acids, triacylglycerols, cholesterol and esterol esters. You will leave the polar fraction (all the phospholipids) on the bottom of the plate.
If using some phospholipids rich source, you could also go for a double development, using methyl acetate/isopropanol/ chloroform/methanol/0.25% (wt/vol) KCl (25:25:25:10:9, by vol) first, to separate the phospholipids. Then you can leave the plate dry and run again, using the neutral mix this time. In that way you could see all lipid classes.
Because you are looking into use oils or butter, I would go for the development using the neutral mix.
Petroleum ether is less pure than hexane, contains hexane but also other hydrocarbon compounds, but it is also cheaper. So I would use the first mix that you suggest if you can't use hexane.
Any thoughts about the dye you are going to use? I have worked with 2′,7′-dichlorofluorescein in methanol, but you would need an UV lamp to see the bands. I also have used 1% iodine in chloroform. That would be more straight forward, because you could see the bands right away.
I hope it helps!
Noemi
For such type of lipids i use 100% chloroform or 100% dichloromethane. Then visualize the separated spots by para anisaldehyde reagent.
Note: Such type of oils mainly contain triacylglycerols which are un separable compounds, so some oils will appear as a single spot.
good answer Naser Jawad Kadhum Naser Jawad Kadhum Harith Rajab Almosawy Dhifaf Zeki
- Badges
- Science topic