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Ultra Performance Liquid Chromatografy
Ultra Performance Liquid Chromatografy
PERFORMANCE
LIQUID
CHROMATOGRAFY
Presented by :
Kaveri Shivaji Aher.
M.Pharm
Quality Assurance Technique
Guided by :
Dr. A.B.Thomas.
UPLC ,Wikipedia
2
Introduction
In 2004 separation science was revolutionized by
‘’Ultra Performance Liquid Chromatography’’ .
To achieve the dramatic change in 1] Resolution
2] Speed
3] Sensitivity
Based on
Fig. no. 1
3
Improvement in
UPLC :
Higher Linear Velocity,
gives lower minima &
less curvature.
Hold for 1min.
Rapid injection ,low
volume.
Auto sampler is
available.
Higher efficiency
higher temperature
reduce the viscosity of
mobile phase.
cooler maintain the
temperature.
decrease in retention
time
Increase in Resolution.
Table. no. 1
4
Fig. no. 4
6
Fig. no. 5
©2012 Waters Corporation
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Principle
UPLC, HPLC both are based on Van Deemter Equation from 1956.
Explain correlation between flow rate and plate height.
H = A + B/ν + Cν
Where,
1] H = HETP ( Height equivalent to theoretical plate)
H= L/N
Where,
L = Length of column
N = No of theoretical plate
Fig. no.6
2] A,B,C are Constant.
A = Eddy’s diffusion Fig. no.7
Fig. no.8
Fig .no.9
10
Efficiency
of L/Particle size
Column
Fig no. 10
11
Case Study
of analysis
of 12
phthalates
by HPLC
& UPLC.
Table no. 3
13
Fig. no. 12
Fig no.13
15
Fig. no. 14
Instrumentation
1.Binary Solvent Manager ( high pressure pump)
- Moves solvent through system .
- Provide steady and ( pulse free) solvent flow.
- Eg :
Solvent High Pressure.
flow rates
a. 1ml /min 103,421kPa
(1034bar,15000
psi)
b. 2ml /min 62.053kPa
(621bar,9000psi
)at reduced
pressure.
Table no.4
Working :
Fig. no. 18
4.Columns
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Column packed with different technology. Table no. 5
Types
1] Evaporative
light scattering
detector :
- incorporate flow
type nebulizer.
- Charged aerosol
detector (CAD)
- less volatile
property than the
mobile phase. Fig. no. 20
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1] Fluorescence detector :
- Multi-channel, multi wavelength fluorescence detector.
- low volume
- high intensity of Hg – Xe lamp in design that minimize
stray light that enhancing the quality of stray light.
- wavelength 200 nm to 800nm excitation rang .
Fig . No.22
H class , H class Bio attribute.
• Based on Acquity UPLC.
• For Biocompatible fluids for biological
molecule.
• True UPLC Performance.
Fig. no. 23
©2012 Waters Corporation7
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Disadvantages :
Higher Back Pressure compare to conventional HPLC.
Decrease the life of column.
Increase the life of column temperature, reduce the back pressure
problem in UPLC.
The particle of less than 2micron are mostly non –regenerable. Have a
narrow use.
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Application :
1. Determination of Particle in Groundwater.
2. Rapid Dose Formulation Analysis.
3. Analysis of Traditional Chinese Medicines.
4. Multi –residue Analysis of Pharmaceuticals in Waste Water.
5. Identification of metabolite.
6. In manufacturing / Quality Assurance(Q.A.) /Quality control.(Q.C.)
7. Impurity Profiling.
8. Method Development /Validation.
9. Forced Degradation Studies.
10. Dissolution Testing.
11. Bioequivalence / Bio analysis study.
12. Toxicity Studies.
13. Analysis of Explosives.
14. Analysis of Contamination in Foodstuffs
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Application
16. Determination of Phytoconstituents .
Principle, Instrumentation, and Applications of UPLC Open Chemistry Journal, 2016, Volume 3
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References :
Waters ACQUITY UPLC Family UPLC Users Meetings Denmark April, 26-27th
Frederic Forini Waters European Headquarters
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References :
Wu T, Wang C, Wang X, Xiao H, Ma Q, Zhang Q. Comparison of UPLC and
HPLC for analysis of 12 phthalates. Chromatographia. 2008 Nov 1;68(9-10):803-
6.
Richard JP. Time-delay systems: an overview of some recent advances and open
problems. automatica. 2003 Oct 1;39(10):1667-94.
Taleuzzaman M, Ali S, Gilani SJ, Imam SS, Hafeez A. Ultra performance liquid
chromatography (UPLC)–a review. Austin J Anal Pharm Chem. 2015;2(6):1056.
www.google.com
Thank You.
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