HPLC and UPLC

Comparison of HPLC Vs UPLC: Which One Is The Best

Table of contents

Introduction and Outcome

HPLC and UPLC are widely used instruments in the pharmaceutical industry. Pharmaceutical development is impossible without these instruments. These instruments are used at every stage (from reaction monitoring to the release of the material). In this article, I will explain the basic differences between these two instruments. After reading this post, all your doubts will be cleared and you will be able to answer several questions:

  • What is the principle of HPLC and UPLC?
  • What are the difference between HPLC and UPLC?
  • What are the limitations of UPLC?

Principle of HPLC and UPLC

Both HPLC and UPLC are used to separate, identify and quantify different components from the sample mixture. The principles for both HPLC and UPLC are the same. Both HPLC and UPLC are separation techniques based on the solid stationary phase and liquid mobile phase. Separation is achieved by partition, adsorption or ion exchange process. Separation is governed by the polarity and chemistry of the stationary phase, mobile phase and molecule.

Differences between HPLC and UPLC

HPLCUPLC
The full form of HPLC is High performance liquid chromatographyThe full form of UPLC is Ultra performance liquid chromatography
It is used to separate, identify and quantify each component of the sample mixtureIt is also used to separate, identify and quantify each component of the sample mixture
Maximum operating pressure is 6000 psiMaximum operating pressure is 15000 psi (higher than the HPLC)
Stationary phase particle size is 2.5 to 5 micronsStationary phase particle size is less than 2 microns
Inner column diameter: 3 to 10mmInner column diameter: 0.75 to 1.8 mm
Run time: longer run timeRun time : shorter run time
Peak shape: Symmetrical and well-shapedPeak shape: More sharp than HPLC
Resolution: goodResolution: better than HPLC
Sensitivity: GoodSensitivity: Better than HPLC
Solvent consumption: moreSolvent consumption: much less than HPLC
Mobile phase flow rate: Generally between 0.5 to 2 ml/minuteMobile phase flow rate: Generally between 50μl to 100 μl/minute

Which one is the best between HPLC and UPLC?

UPLC provides results ten times faster than HPLC. It can tolerate higher pressures due to the smaller particle size in the column compared to HPLC. Secondly, UPLC gives higher resolution, lower tailing factor, higher theoretical plate and sharper peak than HPLC. Therefore, UPLC is better than HPLC.

Applications

HPLC and UPLC are used in both qualitative (like reaction monitoring, identification and purity tests) and quantitative analysis (like assay and content tests) in:

  • Pharmaceutical industries
  • Pesticide industries
  • Food industries
  • Research and development centres and
  • commercial Laboratory
  • Fast analysis
  • Less solvent consumption and
  • High sensitivity

Disadvantages of UPLC

  • Does not work in normal phase chromatographic mode
  • Analysis performed at high pressure

Conclusion

Considering analysis time and consumption of the solvent UPLC is better than the HPLC. We can also say that the UPLC is an advanced version of the HPLC. I hope this post has enhanced your knowledge to the next level. Write your learning and opinion in the comment section.

FAQs

What is the differences between HPLC and UPLC

The major difference between HPLC and UPLC is that HPLC’s maximum operating pressure is 6000 psi whereas in UPLC, the maximum operating pressure is 15000 psi.

What is the principle of UPLC?

UPLC is the separation technique based on the solid stationary phase and liquid mobile phase. Separation is achieved by partition, adsorption or ion exchange process. Separation is governed by the polarity and chemistry of the stationary phase, mobile phase and molecule.

What is the pressure limit for HPLC?

The maximum operating pressure is 6000 psi

What is the pressure limit for UPLC?

The maximum operating pressure is 15000 psi

Are HPLC and UPLC the same?

No. Both are separating techniques but there is difference between HPLC and UPLC. UPLC is the advanced version of HPLC

What is the Typical UPLC flow rate?

Typical flow rate in UPLC is between 50μl to 100 μl/minute

Abbreviations:
  • PSI: pound-force per square inch
  • HPLC: High performance liquid chromatography
  • UPLC: Ultra-performance liquid chromatography
References
  • HPLC Vs UPLC

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