Extraction method (1): Reflux

Used with solid samples, such as hair. Simply placing a sample in a suitable solvent and waiting for any substances of interest to dissolve into the solvent is very inefficient, and would not give good quantitative results. Often, solvent extraction by reflux is used. A suitable solvent is chosen: The substances of interest must be soluble in it. Its boiling point must be low enough not to damage the substances we are extracting. For amphetamines in hair, a mixture of Toluene and Dichloromethane is often used. If we were to heat the solvent (and sample) in an open flask, the solvent would soon evaporate. Refluxing involves heating a solution to boiling and then returning the condensed vapours to the original flask before they are lost by evaporation. A reflux apparatus consists of a round-bottom flask fitted with a reflux condenser. As the solvent in the flask boils, the vapours rise into the condenser where they are converted back to a liquid that drips into the original flask. A typical reflux is run for at least one hour.

Extraction method (2): Solvent extraction

Used with aqueous samples, such as urine. Here we want to extract the substance of interest from an aqueous medium into an organic solvent. Again, we need to be careful about choosing a suitable solvent: It must be immiscible in water The substance of interest should be more soluble in it that in water. Ideally, they should have different densities. If you mix (for example) Dichloromethane and urine, you will see that they separate into two distinct layers they are said to be immiscible. In this case Dichloromethane is denser than water and will form the bottom layer. Solvent extraction is done using a separating funnel. This is a glass tube with a tap at one end and a stopper at the other. If we vigorously shake the funnel (with tap closed and stopper tightly fitted!), the two layers will mix and the compound we are interested in will now be distributed between the two layers. Depending on the system in use, there will be an upper limit to the amount we can extract into the organic layer. This can be quite high, say 90%, but will never be 100%. So how do we improve the efficiency of the extraction? We simply run off the organic layer, add some more organic solvent, and then repeat the extraction. Again our 90% limit applies, but the after first stage we only had 10% left, so the second extraction extracts 90% of the remaining 10%. So, after only two extractions we have a 99% efficient extraction.

Removal of extraction solvent using a rotary evaporator

After using either extraction method, we are left with our sample in a fairly large volume of an organic solvent. If we want to find out how much sample we have by weighting it, or we want to concentrate the solution for analysis, we have to remove most of the solvent. This is normally done using a rotary evaporator. This is a sophisticated distillation system employing: Reduced pressure (solvents evaporate at lower temperature at lower pressures) A rotating spindle which turns the boiling flask in the water bath (this reduces the chances of local hot-spots forming) A solvent recovery system.