I suggest you to analyze the total lipid fraction (without transesterification reaction) by using capillary column RTX 65-TG (30 m; 0.25mm ID; 0.1 ft; Restek) and applying a temperature programme starting from 50°C; T increase of 10°C/min; final temperature 360°C; hold for 5 min. In this way you will be able to identify many lipid categories like triglycerides, wax esters, sterols, steryl esters, diglycerides.

I suggest you to analyze the total lipid fraction (without transesterification reaction) by using capillary column RTX 65-TG (30 m; 0.25mm ID; 0.1 ft; Restek) and applying a temperature programme starting from 50°C; T increase of 10°C/min; final temperature 360°C; hold for 5 min. In this way you will be able to identify many lipid categories like triglycerides, wax esters, sterols, steryl esters, diglycerides.

Thank you. That is very helpful.

Besides, after and before capillary column, you can use thin layer chromatography to qualitatively estimate "purity" depending on solvent eluent polarity. And hence use NMR spectroscopy to integrate certain types of compounds of your interest, such as Prof. Naviglio stated, e.g. triglycerides, waxy esters, fatty esters and so on. Just revise 1H NMR of those present compounds in order to track them in the spectra. Have luck in your determinations.

HPTLC is good option for you.

Lipids can be conveniently analysed by HPTLC /HPLC . You may review the following ref.

1.Extraction of Lipids from Tissues - Lipid Library

lipidlibrary.aocs.org/topics/extract2/file.pdfFile

by WW Christie -

extract lipids more thoroughly from animal, plant or bacterial tissues than other simple solvent combinations (the endogenous water in the tissues should be ...

2. Determination of Lipids in Animal Tissues - University College London

www.ucl.ac.uk/~ucaptt0/TLC.

Densitometrie HPTLC Determination of Lipids in Animal tissues were extracted as soon as possible after animals had been killed. When this was not feasible the ...

3. of lipid extraction, methylation and gas chromatography, for the ...

editorialbiogenesis.udea.edu.co/index.php/biogenesis/..

by J Angulo - 2011

Lipid extraction and methylation . .... procedures for lipid extraction from animal tissues will be .... for HPLC and 500μL of chloroform/methanol (2/1) if are going to ...

Lipid extraction by methyl-tert-butyl ether for high-throughput ...

if you extract by organic solvents, try extraction by NaHCO3 5% and then nutralization by HCl.

or you can analyze extraction br GC-Mass with an appropriate column and identification your mixture by comparing sample owadays GC-Mass instruments equipped by different large libraries. Good Luck

try to do dupe solvent extraction using HPLC grade solvent. Then inject the sample to LC-MS using appropriated column. this process will take less than 10 minutes.

I dont have many experience with HPLC/LC systems. Im guessing I would have to use a normal phase column. Which would be "an appropriated column" for my type of samples (lipids, with intermediate to high polarity compounds)?

I think gc/gc-ms and tlc techniques are useful to confirm the purity of ur extract, no doubt nmr is best technique but is time consuming.

Which methods are established in your lab (TLC, GC, NMR, HPLC...; detection: MS, UV...). Depending on the lipid class you are investigating I would also recommend TLC. TLC is an easy way to get qualitative information about the purity of your sample since you can seperate and detect a broad range of lipid classes in a single run. Moreover "fatty acids containing lipids" like glycero- or glycerophospholipids can be easily isolated by TLC and used for further analysis. Otherwise HPLC techniques (e.g. reversed phase HPLC/MS) are the more sophisticated approaches but this depends on the facilities of your lab. Good luck

If you have enough of the extract (5-20 mg), the best way to get the whole picture is to measure NMR. Simple 1H proton spectrum shall tell you the amount of glycerolipids, waxes, alcohols, fatti acids, degree of unsaturation, average chain length, amount of sterols. Combined with TLC mobilities and GC data this gives quite much of information.

1- On your bench, you can develop a TLC (thin-layer chromatogram) using an appropriate solvent system. It is a cheap and self-dependent strategy.

2- If you have facilities of Mass Spectrometry or NMR then you can use both of them to check the purity.

3- you can also use the capillary column suggest by Daniele Naviglio.