HPLC INSTRUMENTATION / What is HPLC?
The figure above shows a typical HPLC system.
acronym for "High Performance Liquid Chromatography", saw widespread
adoption beginning around 1970 with the advent of high-pressure
"Chromatography" refers to "compound separation", a
technique for separating the components contained in a mixture to
understand their properties and quantities. The crux of compound
separation is the "separation column". The stationary phase within
the column interacts with compounds, and differences in the
interaction power result in variations in elution time (i.e.,
An HPLC system is primarily composed of three
The mobile phase (the liquid
moving within the column) is drawn from the reservoir by the pump
and discharged, providing stable delivery to the column at a set
When a few to several
hundred μL of sample solution containing solutes (in the case of
analysis) is mixed into the mobile phase and "injected" into the
column, a phenomenon occurs where the solutes interact on a
molecular level with the stationary phase within the column,
resulting in retention and separation.
When the solutes separated and eluted from the column pass through
the detector, they exhibit qualitative and quantitative responses.
These responses are converted electrically, recording digital
signals such as time-response or spectra.
In recent years,
HPLC, with a mass spectrometer (MS) as the detector in the form of
LC-MS, has become an indispensable tool for chemical analysis. It is
used for identifying and quantifying trace elements in various
fields such as pharmacology, food, environmental studies, and