Comparison of HPLC Vs UPLC: Which One Is The Best
Table of contents
- Introduction and Outcome
- Principle of HPLC and UPLC
- Differences between HPLC and UPLC
- Which one is the best between HPLC and UPLC?
- Advantages of UPLC
- Applications
- Conclusion
- Disadvantages of UPLC
- FAQs
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
| HPLC | UPLC |
| The full form of HPLC is High performance liquid chromatography | The full form of UPLC is Ultra performance liquid chromatography |
| It is used to separate, identify and quantify each component of the sample mixture | It is also used to separate, identify and quantify each component of the sample mixture |
| Maximum operating pressure is 6000 psi | Maximum operating pressure is 15000 psi (higher than the HPLC) |
| Stationary phase particle size is 2.5 to 5 microns | Stationary phase particle size is less than 2 microns |
| Inner column diameter: 3 to 10mm | Inner column diameter: 0.75 to 1.8 mm |
| Run time: longer run time | Run time : shorter run time |
| Peak shape: Symmetrical and well-shaped | Peak shape: More sharp than HPLC |
| Resolution: good | Resolution: better than HPLC |
| Sensitivity: Good | Sensitivity: Better than HPLC |
| Solvent consumption: more | Solvent consumption: much less than HPLC |
| Mobile phase flow rate: Generally between 0.5 to 2 ml/minute | Mobile 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
Advantages of UPLC
- 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.
You may also want to check out other articles on my blog, such as:
- Difference between HPLC and GC
- How to develop a method using GC-MS?
- Need of Chromatographic Method in Drug Development
- What is the Chiral Gas Chromatography?
- How to decide system suitability test (SST)?
- Allowable GC Method Adjustment
- How to develop the HPLC method for basic compounds?
- How to develop the HPLC method for acidic compounds?
- How to develop the HPLC method for non-polar compounds?
- What should be the Analytical method development approach?
- LC-MS in pharmaceutical development
- Tips on HPLC column equilibration and
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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