UPLC refers to ultra performance liquid chromatography. It enhances speed, resolution, and sensitivity compared to HPLC by using particles less than 2μm in diameter. UPLC operates at very high pressures and provides better separation and faster analysis. It has applications in determining pesticides, analyzing pharmaceutical impurities, and more. UPLC offers advantages like reduced run time and solvent usage but also has disadvantages like higher back pressures reducing column life.

1. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 1
A Seminar as a part of curricular requirement for I year M. Pharm I semester
Presented by
P. Sudheer Kumar (20L81S0712)
M. Pharmacy
Department of Pharmaceutical Analysis, RIPER.
Under the Guidance of
Dr. P. Ramalingam, M. Pharm., Ph. D
Director – R & D
Professor of Pharmaceutical and Medicinal Chemistry.
Ultra Performance Liquid
Chromatography

2. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 2
• Introduction of UPLC
• Principle
• Applications
• Advantages
• Disadvantages
• Instrumentation
• HPLC vs UPLC
• Conclusion
• References
Contents

3. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 3
• UPLC refers to ultra performance liquid chromatography, which
enhance mainly in three areas: speed, resolution and sensitivity.
• UPLC is applicable for particle less than 2μm in diameter to acquire
better resolution, speed, and sensitivity compared with HPLC.
• The separation and quantification in UPLC is done under very high
pressure (up to 100M Pa).
• In this separation mechanism the principle involved is Van Deemeter
equation.
Introduction

4. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 4
• It evaluates the efficiency as a function of linear viscosity or flowrate.
• HETP- Height equivalent to theoretical plate.
• L- Column length, N- Number of theoretical plates.
Van Deemter equation

5. RIPER
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NAAC &
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SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 5
• Smaller particles
-Provide the increase efficiency
-Maintain efficiency over a wider linear velocity
-Provide the ability to add resolution and increase the speed of separation
• Particles are central to the quality of separation

6. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 6
• Determination of pesticides in ground water
• Improved resolving power in peptide maps
• Rapid dose formulation studies
• Multi residue analysis of pharmaceuticals in waste water
• Impurity profiling
• Forced degradation studies
• Analysis of contaminants in food stuff
• Analysis of explosives
• Toxicity studies
• Therapeutic drug monitoring
Applications

7. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 7
• Require less run time and enhance sensitivity.
• Provides the selectivity, sensitivity, and dynamic range of LC
analysis.
• In chromatogram resolved peaks are obtained.
• Speedy analysis, accurately quantifies the analyte and related
products.
• Use of fine particle (2μm) for packing of stationary phase make
analysis fast.
• Time and cost both are reduced.
Advantages of UPLC

8. RIPER
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NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 8
Contd…
• Consumption of solvents is less.
• More products are analyzed with existing resources.
• Delivers real-time analysis in step with manufacturing processes.
• Assures end-product quality, including final release testing.

9. RIPER
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NAAC &
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SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 9
• The higher back pressures decreases the life of the columns.
• Increasing the column temperature reduces the back pressure problem
in UPLC.
• Moreover, the particles of less than 2 μm are mostly non-regenerable
and therefore, have a narrow use.
Disadvantages of UPLC

10. RIPER
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NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 10
Instrumentation of UPLC

11. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 11
S.No. UPLC HPLC
1. Particle size of stationary phase is less
than 2 microns (typically 1.7 microns).
Particle size of stationary phase is in
between 3 to 7 microns.
2. Operates at high pressure due to reduced
particle size.
Operates at relatively low pressure
than UPLC.
3. Maximum back pressure of 1000
bars/100 Mpa.
Maximum back pressure between 300
to 400 bars.
4. Inner diameter of the column is generally
0.75- 1.8 mm.
Inner diameter of the column is
generally 3- 10 mm.
5. Reduces process cycle time and assures
end product quality with reduced cost of
operation and decreased run time.
High cost of operation and high run
time as compared to UPLC.
HPLC vs UPLC

12. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 12
S.No. UPLC HPLC
6. It decreases the consumption of solvent
and increases the solvent throughput.
More sample consumption, less
sample throughput.
7. The higher backpressure compared to
HPLC decreases the life of the columns
(column coagulation). Increase the
column temperature reduces the back
pressure problem in UPLC.
Less back pressure, more life of the
column.
8. Injection volume are small,
generally 2 microliter.
Injection volume are large,
generally 5 microliter.
9. Less run time. More run time.
HPLC vs UPLC

13. RIPER
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SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 13
S.No. UPLC HPLC
10. Systems should withstand high pressures,
more efficient pumps are required.
Less efficient pumps are required.
11. Stationary phase is modified to withstand
high pressures. Hybrid of inorganic silica
and organic group is used. Eg- Bridged
ethylene hybrid (BEH), High strength
silica (HSS), Charged surface hybrid (CSH)
Inorganic silica modified to different
polarity are used.
12. Low extra column dispersion. More extra column band broadening.
13. More plate count is achieved during
analysis.
Less plate count is achieved during
analysis.
HPLC vs UPLC

14. RIPER
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SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 14
HPLC vs UPLC

15. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 15
UPLC is one of the most important tools in analytical chemistry
which increases the speed, resolution, and sensitivity of the
chromatographic analysis and decreases the time, solvent
consumption and cost involved. This technology thus creates a new
opportunity for business profitability in highly efficient manner and
allows the product to be introduced to the market in less time.
Conclusion

16. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 16
• Taleuzzaman M, Ali S. Ultra Performance Liquid
Chromatography- A Review. Journal of Analytical and
Pharmaceutical chemistry. 2015; 2(6): 1056.
• Gita chawla. Principle, Instrumentation and Applications of UPLC:
A novel technique of liquid chromatography. Open Chemistry
Journal. 2016; 8: 1874-8422.
• Srinivas rao. S. Ultra Performance Liquid Chromatography- A
Review. Research Gate. 2020; 11(3): 8051-8056.
References

17. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 17