Chemical detection of Aflatoxins in dairy products

High performance liquid chromatography

High performance liquid chromatography (HPLC) is a widely used analytical technique. It allows samples to be separated and their components determined.

The basic principle of HPLC is passing a sample through a column packed with tiny particles (usually less than 10um diameter) and applying high pressure to increase the resolution and speed of the technique.


There is solvent constantly flowing through the system. This solvent is called mobile phase. To ensure the solvent is constantly moving through the system a pump is employed. This gives the required pressure to allow mobile phase and sample to move through the column with ease and speed. The sample is injected into the mobile phase so it can proceed directly to the column. The sample comes out from the column (elutes) and proceeds to the detector. This monitors the refractive index difference between the pure mobile phase entering the column and the column elute.

The signal from the detector is sent to a data recorder. This measures the time of each component (retention time) and the size of the signal (peak area).
By injecting a known standard a retention time and peak area are achieved. If an unknown sample is then injected and peak obtained at the same retention time.
The peak area can be used to calculate the amount of standard material in the unknown by proportion.

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