Difference Between HPLC and UPLC


HPLC and UPLC are both Liquid Chromatography used in separating the components of a compound. To know the difference between HPLC and UPLC, we first need to know what HPLC is. This is because UPLC is a special version of HPLC and can be easily understood if we know what HPLC is. For starters, HPLC stands for High Performance Liquid Chromatography is a technique used to separate different constituents of a compound. It is the most widely used technique to identify, quantify and separate components of a mixture. It uses high pressure to push solvents through the column. HPLC is widely used in biochemistry for the analysis of constituents of a compound. It is an ideal way for separation and identification of amino acids, nucleic acids, proteins, hydrocarbons, pesticides, carbohydrates, antibiotics, steroids and countless other inorganic substances.

HPLC has many advantages over previously used liquid chromatography methods. It allows for higher resolution and speed of analysis, HPLC columns can be reused without repacking or regeneration, better control of parameters affecting efficiency of separation, easy automation of instrument operation and data analysis, and adaptability to large scale procedures. This is also because it supports very small particle sizes and large surface areas and allows for application of high pressure for solvent to flow.

In UPLC, which is nothing but ultra high performance liquid chromatography, particle sizes less than 2um can be used. This allows for better separation that particle size of 5um that are used in HPLC. It also allows for a very fast analysis. Though, UPLC is a trademark technology of Waters Corporation, people use it as a general term for a separate technique. Where as the pump pressure in HPLC is 40MPa, in UPLC, this pressure can go up to 100MPa, which is what makes this technique so very exciting and efficient.


• HPLC is high performance liquid chromatography, whereas UPLC is ultra high performance liquid chromatography

• In essence, UPLC is just a special variant of HPLC

• The pump pressure, which is 40Mpa (400 atmospheres) in HPLC, can go up to 100MPa in UPLC. UPLC also allows for much smaller particle size, normally less than 2um whereas it is normally 5um in HPLC.