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Current Trends in UPLC

This document discusses current trends in ultra-performance liquid chromatography (UPLC). UPLC uses fine particle columns and high pressures to improve chromatographic resolution, speed, and sensitivity compared to HPLC. It works on the principles of adsorption or partition chromatography. The instrumentation includes binary solvent managers, sample managers, column heaters, pumps, and detectors like UV and ELSD. UPLC has applications in drug discovery, quantitative analysis, dosage form analysis, amino acid analysis, and pesticide residue detection. Advantages are faster analysis times, higher sensitivity, and reduced solvent usage. Disadvantages include higher maintenance needs and non-regenerable columns under 2 micrometers.

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Diwya Kumar
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46 views15 pages

Current Trends in UPLC

This document discusses current trends in ultra-performance liquid chromatography (UPLC). UPLC uses fine particle columns and high pressures to improve chromatographic resolution, speed, and sensitivity compared to HPLC. It works on the principles of adsorption or partition chromatography. The instrumentation includes binary solvent managers, sample managers, column heaters, pumps, and detectors like UV and ELSD. UPLC has applications in drug discovery, quantitative analysis, dosage form analysis, amino acid analysis, and pesticide residue detection. Advantages are faster analysis times, higher sensitivity, and reduced solvent usage. Disadvantages include higher maintenance needs and non-regenerable columns under 2 micrometers.

Uploaded by

Diwya Kumar
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PPTX, PDF, TXT or read online on Scribd
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CURRENT TRENDS IN

UPLC
Presented by:
Diwya Kumar Lal
7th Sem/ B. Pharmacy
Contents:
 INTRODUCTION

 RECENT ADVANCEMENTS

 PRINCIPLE

 INSTRUMENTATION AND WORKING

 APPLICATIONS

 ADVANTAGES

 DISADVANTAGES
INTRODUCTION
 Ultra-Performance Liquid Chromatography (UPLC) is a relatively new
technique giving new possibilities in liquid chromatography, especially
concerning decrease of time and solvent consumption

 It improves in three areas: chromatographic resolution, speed and


sensitivity analysis.

 It uses fine particles and saves time and reduces solvent consumption.
RECENT ADVANCEMENTS
 As compared to HPLC, the separation on UPLC is performed under
very high pressures (up to 100 MPa) but it has no negative influence on
analytical column or other components of chromatographic system.

 It also facilitates the analysis of complex mixtures in relatively short


time and the peaks obtained with the help of these method provides
more information which is more convenient and clearer that of HPLC.
PRINCIPLE:
The principle for the separation can either be adsorption or partition.

Adsorption chromatography:
 When adsorbate or mixture compounds is dissolved in mobile phase (eluent) travels
through a column of stationary phase (adsorbent), they move according to the relative
affinities towards stationary phase.

Partition chromatography:
 The most widely used type of UPLC is partition chromatography.
 The early forms of Partition chromatography liquid-liquid columns. These have been
replaced in modern LC systems by liquid – bonded phase columns.
 In liquid – liquid chromatography, the liquid was held in place by physical adsorption.
In bonded – phase chromatography, its attached by Chemical Bonding, resulting highly
stable pickings insoluble in the mobile phase.
INSTRUMENTATION AND WORKING:
The various instruments used in the Ultra performance liquid chromatography are
as follows:

 Binary solvent manager

 Sample manager including the column heater

 Optional Sample manager

 Pumps

 Detector
INSTRUMENTATION AND WORKING:
INSTRUMENTATION AND WORKING:
Binary Solvent Manager:
 The binary solvent manager uses two individual serial flow pumps to deliver a
parallel binary gradient. The binary solvent manager is a high-pressure pump that
moves solvent through the system.
 It provides steady (pulse free) solvent flow at analytical flow rates.

Sample Manager:
 The Acuity sample manager injects the sample it draws from Micro titer plates or
vials in to the chromatographic flow stream. A locating mechanism uses a probe to
access sample locations and draw sample from them.
 The Sample manager can perform an injection in approximately 15 seconds.
INSTRUMENTATION AND WORKING:
Column Heater
The column heater is of a modular design and its foot print is identical to that of the
sample manager. Thus, it attaches to the top of the sample manager and serves as that
instrument’s top cover.

Optional Sample Organizer


The optional sample organizer stores micro miter or vial plates and transfers them to
and from the sample manages, automating their processing and increasing through put.
INSTRUMENTATION AND WORKING:
Pumps:
 The UPLC pump is considered to be one of the most important components in a
liquid chromatography system which has to provide a continuous constant flow of
the eluent through the UPLC injector, column, and detector. The two basic
classifications are
 Constant pressure pump: The constant pressure is used only for column packing.
 Constant flow pump: This type is mostly used in all common UPLC application.

Detectors:
Detectors can be classified as:
 Optical detectors
 Tunable ultra violet detectors
 Evaporative light scattering detectors
APPLICATIONS

 1. Drug Discovery: UPLC improves the drug discovery process by means of high
throughput screening, combinational chemistry, high throughput in vitro
screening to determine physiochemical and drug’s pharmacokinetics.
 2. High throughput quantitative analysis: UPLC coupled with time of flight
mass spectroscopy give the metabolic stability assay.
 3. Analysis of Dosage form: It provides high speed, accuracy and reproducible
results for isocratic and gradient analysis of drugs and their related substance.
Thus method development time decrease.
 4. Analysis of amino acids: UPLC used from accurate, reliable and reproducible
analysis of amino acids in the areas of protein characterizations, cell culture
monitoring and the nutritional analysis of foods.
 5. Determination of Pesticides: UPLC couples with triple Quadra-pole tandem
mass spectroscopy will help in identification of trace level of pesticides from
water.
ADVANTAGES:
 It decreases the run time and increases sensitivity.

 Provides the sensitivity, selectivity, and dynamic range of LC analysis.

 Maintains the resolution performance.

 Expands scope of Multiresidue Methods.

 Faster analysis is possible with the use of a novel separation material which
are of very fine particle size.

 The cost of operation in less.

 Solvent consumption is less.


DISADVANTAGES:
 Due to increased pressure requires more maintenance and reduces the life of
the columns of this type.
 So far performance similar or even higher has been demonstrated by using
stationary phases of size around 2 μm without the adverse effects of high
pressure.
 In addition, the phases of less than 2 μm are generally non-regenerable.
THANK YOU

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