How to evaporate choloroform off from lipid suspension - (Oct/01/2007 )

Do someone know how to evaporate chloroform off from the fatty acid dissoved in chlorofrom
I has tried by using negative pressure (25 mmHg for 5 hrs) however less than 20 % of chloroform evaporated
If someone know the good method please tell me i would be appreciate.

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I dont know much about lipid suspension, but does it work if you just leave it under fume hood overnight?

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Try drying under nitrogen. What vials or tubes are you using? You might need to attach a glass pipette to the end of your gas supply and suspend that right down inside the test tube.

Also, try using organic phases other than chloroform. Would di-ethyl ether not work? Also, we acidify samples first to prevent ionization of the fats.

Hope that helps.

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If you meant residual chloroform in lipid, you need to remove it using an oil pump for overnight. Otherwise blow it with N2 stream or rotovap for initial removal of the bulk chloroform solvent.

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Drying under N2 is the best way to avoid oxidation of your fatty acids.
Your problem maybe due to high amount of lipids. Use more solvent and bigger glass tube.

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My total volume is just 500 uL of Chloroform as a solvent to mix 3 fatty acids in eppendoft tube.
How could i know that all chlorofrom completly evaporated.
Are there any method to test this?

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classical protocol with Argon; N2 only if there are no polutions with O2

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I would suggest you to use glass culture tubes instead. You dry it under N2, then vacuum till the weight is constant.

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Dear all,

My name is Joana, and I am currently starting to work with lipids.
I use Drosophila as a model system.

Since I am new to this field, I am having some difficulties in getting enough good quality lipid extracts in my samples.

Does anyone knows a protocol suited for experiments using Drosophila tissue?

Most protocols I see use the Folch or Bling Dyer Methods.
In these protocols, after concentration of the sample (by speed vac), in what solvent do you ressuspend your lipids? (I will be quantifying the lipids using colorimetric methods, so if I can avoid chloroform or methanol, that would be excelent, since the 96-well plates I have are not resistant to ant of these solvents).
Also, does anyone knows if this "concentration" step be done just by "air-drying" (I do not have a speed vac)?

Thanks,
Joana

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If nitrogen or argon are not available in your lab, you can add butylated hydroxytoluene (e.g. 0.01%) to the samples to avoid oxidation. Then you can use a stream of air to dry the samples

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