HPLC - Glossary - peptidetesting.cc

HPLC is an analytical chemistry method that separates molecules in a liquid sample based on their chemical properties. This technique produces a chromatogram, which is a graph used in lab reports to record and interpret the substances detected during testing.

The HPLC separation process uses high pressure

The HPLC process works by pumping a liquid solvent at high pressure through a column packed with solid material. When a sample is injected, its components interact with the solid material in different ways, causing them to move at different speeds. As each component exits the column, a detector measures it to create the final chromatogram.

Confirming identity: The time a substance takes to exit the column is compared to a known standard to verify its identity.
Measuring purity: The detector compares the target compound against impurities to generate an area-percent purity score, such as 98% pure. This indicates the sample contains very few unwanted byproducts or degradation materials.

HPLC and Mass Spectrometry serve different roles

HPLC is often confused with Mass Spectrometry (MS) in lab reports, but the two methods provide different information. HPLC determines purity by measuring how much of a substance is present, while Mass Spectrometry identifies the substance by its molecular weight. Because HPLC alone can miss hidden impurities, labs often combine both methods into a single LC-MS test.

The HPLC separation process uses high pressure

HPLC and Mass Spectrometry serve different roles

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