Difference between HPLC and UPLC


HPLC meaning : High Performance Liquid Chromatography or High Pressure Liquid Chromatography.

UPLC meaning: Ultra-performance liquid chromatography

What is HPLC (High Performance Liquid Chromatography)?

  • HPLC is known as High-Performance Liquid Chromatography, which is previously denoted as high-pressure liquid chromatography.
  • For the past 30 years, HPLC has been one of the most dominating and commonly utilised technologies in analytical laboratories.
  • Pressure: HPLC system can operate at pressure up to 6000 psi.

Realated: Calibration of HPLC, Difference Between Thin Layer and Paper Chromatography, Type of HPLC Column

HPLC (High Performance Liquid Chromatography)
Waters HPLC

What is UPLC (Ultra Performance Liquid Chromatography)?

  • UPLC Definition:  UPLC’s full name is Ultra Performance Liquid Chromatography. Because of its ability to increase sample throughput, chromatographic efficiency, sensitivity, and reduce run time, UPLC chromatography has become the modern standard HPLC platform.
  • Pressure: UPLC system can operate at pressure up to 15,000 psi.
UPLC (Ultra Performance Liquid Chromatography)
Credit@ Waters Corporation, USA

What is the Difference Between UPLC and HPLC?

  • The liquid chromatography techniques HPLC and UPLC are both used to separate the different components contained in mixtures.
  • UPLC, on the other hand, uses higher pressures (15,000 psi) and allows for smaller particle sizes in the columns, whereas HPLC uses lower pressures (max 6000 psi).
  • Particle sizes: UPLC has a stronger sensitivity as well as the ability to handle higher pressure levels. As a result, it is suitable for low-component analysis since it allows for smaller particle sizes within columns.
  • UPLC improves analyte resolution and sensitivity while also reducing solvent usage and reducing run times.
  • UPLC is a common laboratory technique that decreases the cost of designing and validating a method while increasing the efficiency of analysis.

Brief on HPLC vs UPLC:

Sr. No.ParameterHPLCUPLC
 1HistoryThe work of Russian botanist Mikhail S. Tswett defined liquid chromatography (LC) in the early 1900s.  
In 1967, Waters Associates (now Waters Corporation), a business that provides analytic tools and systems to scientists, developed the first commercially available HPLC system, the ALC100 HPLC.
Waters Corporation USA, in the year 2004, launched and trademarked Ultra Performance Liquid Chromatography (UPLC).
 2MeaningHigh Performance Liquid ChromatographyUltra-performance liquid chromatography
 3Operating Pressure15,000 psiLess than 6000 psi
 4Stationary Phase– Particle Size3 to 5µmLess than 2µm
 5Injection Volume5-20 µLGenerally, 2µL
 6Column Inner Diameter0.75 to 1.8 mm3 to 10 mm
 7Dimension of Column 3.9 to 4.6 mm id and 15.0, 25.0 & 30.0 cm in lengthGenerally, 150mm L* 2.1 mm  id  
Selective & SensitivityLess than UPLCMore than HPLC
 9ResolutionLess than UPLCMore than HPLC
 10Retention timePeak retention time is more  Reduce process cycle time and assures great quality.
 11Solvent ConsumptionMore solvent consumption so, More cost than UPLCLess solvent consumption than HPLC so, Less cost of operation
 12Back Pressure300-400 bars Relatively less backpressure than UPLC1000 bars High Backpressure leads to decrease column life.
 13Theoretical Plat countLess Theoretical Plat count than UPLCMore Theoretical Plat count

What are the differences between FPLC and HPLC

  • FPLC full name is Fast protein liquid chromatography (FPLC), is a method of liquid chromatography that is frequently used to analyze or purify mixtures of proteins.
  • FPLC was developed and promoted in Sweden by Pharmacia in the year 1982, and was originally called fast-performance liquid chromatography (FPLC) to difference it with HPLC (High-performance liquid chromatography).
  • Although FPLC is typically used to separate proteins, it has a wide range of applications due to the variety of resins and buffers available.

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