Question

How could you use HPLC to determine the purity of your acetaminophen sample?

Answer 1

HPLC --> stands for high performance liquid chromatography; which is used in determination of solvents

similar to the TLC; which stands for Thin Layer Chromatography; it is an analytical technique in which we exploit each sample's affinity toward different polar/nonpolar solvents.

We use a stationary phase: a phase which does NOT moves, as the name implies. It is typically the "thin layer" or the "plate". It is commonly to use silica gel plates, which are polar, but any species could be used, such as paper.

We also use a mobile phase: as the name implies, this phase will "move" and will carry along the path all the components from the solvent. Many times, you will be able to split in several components the "mixture" of the sample. Therefore, they can be seen in the stationary phase (plate). Typically, we allow the mobile phase to go up to the "Rfinal" mark, or final reference. This is the distance travelled by the solvent (mobile phase). It starts at solvent level, it is R0, therefore.

Rf = distance traveled by sample / distance traveled by solvent

Then, "Rf" value allows us to understand the affinities of anaytes towards the solvent.

As Rf increases, this implies higher affinity toward the mobile phase ( since it is carried all along the mobile phase)

If Rf is low, this implies a low affinity toward the mobile phase; therefore higher affinity toward the stationary phase, since it sticks to the plate rather than travel upwards.

now... for the impurity of the acetominophen:

Recall that acetominophen is a pure substance and so impurities can be considered a mix of 1 or more pure susbtance

each impurity will have a different retention towards solvent/column

therefore, we can separate acetominophen's from the impurities and identify the different species present in the plate/column

this way, we would expect 1 spot if pure, and at least 2spots if acetominophen + impurities