Wednesday, June 16, 2021

Difference between HPLC and UPLC

The major difference between HPLC and UPLC is that due to particle size HPLC works at lower pressures, while UPLC works at higher pressures, which allows UPLC a reduced run time, lower solvent consumption, and better analysis separation and detection by the detector.

Chromatography is a technique used to separate chemical compounds based on differences in their relative affinity for the mobile phase (gas/liquid) and stationary phase (solid/liquid). The different types of chromatography are paper chromatography, thin-layer chromatography (TLC), column chromatography, partition chromatography, high-performance liquid chromatography, and gas chromatography are commonly used for qualitative as well as quantitative determination of molecules.


Both high-performance liquid chromatography and ultra-performance liquid chromatography are the separation techniques used for separate different components found in the complex mixture. UPLC works on high pressures and particle size of columns is less, Whereas, HPLC works under low pressure, hence UPLC offers better detection and separation, shorter run times, and consumption of less solvent.

What is UPLC?

UPLC stands for ultra-performance liquid chromatography, it becomes the modern standard HPLC platform. Because it offers increased efficiency, sampling throughput, sensitivity, and reduced run time making it more powerful and time and cost-effective.

HPLC vs UPLC
Difference between HPLC and UPLC

What is HPLC?

HPLC stands for high-performance liquid chromatography, it is an analytical technique used to separate, identify, and quantify components in a sample mixture. It is the most common chromatography technique used in most laboratories across the world. HPLC system can withstand pressure up to 6000 PSI. The most commonly used form of HPLC is reversed-phase (RP-HPLC).

Difference between UPLC and HPLC:

  • UPLC works on about max 15000 psi pressure, while HPLC works on max up to 6000 psi pressure.
  • UPLC the particle size of the column stationary phase is less than 2 µm, whereas the particle size of the column stationary phase in HPLC is 3 to 5 µm.
  • The one more difference between HPLC and UPLC is they can analyze the sample of UPLC very quickly, which is about 3-10 minutes which can reduce the cost of operation; while HPLC can take about 10-20 minutes to separate the analytes.
  • UPLC analysis required less solvent and increases sample throughput than HPLC, while high-performance liquid chromatography analysis required more solvent or mobile phase and less sample throughput.
  • UPLC is more selective and sensitive than HPLC, while HPLC is less selective and sensitive than UPLC.
  • UPLC has high resolving power compared to HPLC, while HPLC has less resolving power compared to UPLC.
  • Because the packing material particles are less than 2 μm, UPLC columns are often non-regenerable and have a narrow use, whereas HPLC columns have more use.
  • Sample injection necessitates high precision in UPLC, while HPLC has lower comparative to UPLC.
  • A small injection volume is required for UPLC (generally 2 microlitres), while a large injection volume is required for HPLC (generally 5 to 20 microlitres).
  • UPLC system works with higher pressures and requires more efficient pumps than HPLC systems.
  • UPLC achieves a higher theoretical plate count than HPLC, while HPLC achieves a lower plate count.
  • Higher back pressure shortens column life in UPLC, while lower back pressure increases column life in conventional HPLC.

Commonly asked questions on liquid chromatography are as follows.

Which one is more popular between UPLC and HPLC?
In the analytical field, UPLC is more popular between UPLC and HPLC, especially for complex sample analysis, because of its advantages of higher resolution, faster processing speed, and better potential sensitivity.

What is the advantage of UHPLC?
Improved resolution, faster analysis (up to 10 times), eluent saving, enhanced sensitivity, and excellent peak shape is the major advantage of UHPLC.

Why UPLC is higher efficiency than HPLC?
Because of particles less than 2m in diameter ultra-performance liquid chromatography (UPLC) provides superior resolution and higher efficiency than HPLC.


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