Ultra Performance Liquid Chromatography (UPLC) is an advanced analytical technique that enhances the resolution, speed, and sensitivity of liquid chromatography through innovations in instrumentation and particle size. UPLC operates at higher pressures and uses smaller particle sizes compared to High Performance Liquid Chromatography (HPLC), allowing for significantly reduced analysis time without loss of efficiency. This technology is widely applied across various industries, including pharmaceuticals and environmental analysis, providing advantages in cost, quality, and reproducibility compared to conventional methods.

1. ABSTRACT
INTRODUCTION
APPLICATIONS
PRINCIPLE
UPLC is an advance analytical technique where it takes
advantage of innovation in various technologies such as
instrumentation and particle size to achieve dramatic
increases in resolution, speed and sensitivity of the liquid
chromatography. It operates at higher pressure than that used
in HPLC and uses fine particles (less than 2.5µm) & mobile
phases at high linear velocities. It can be hyphenated with
other techniques such as (MS), (IC), (NMR) and (IR) etc. This
is used in all industries and has found application in various
fields such as pharmaceutical, food, environmental, forensic,
toxicology and pesticide.
UPLC V/S HPLC
The Basic principle and instrumentation of UPLC system remains same as that of HPLC but
only differs in upgrading instrumentation and hardware. The UPLC System consists of a
binary solvent system, sample manager, column manager and detector. The solvent
manager uses two flow pumps to deliver a parallel binary gradient which is mixed under high
pressure. Degassing system degasses the mobile phase, which is selected by valve up to four
solvent. UPLC system can withstand pressure of about 15,000 psi (about 1000 bar) to take full
advantage of the sub-2-mm particles. The sample manager is also having advance technology
where sample temperature can be taken to as low as 0°C , whereas column manger can
manage the column temperature up to 90°C using high temperature ultra performance liquid
chromatography (UPLC), it is possible to drastically decrease the analysis time without loss in
efficiency.
From past 30 year HPLC is the predominant technique to use
for analysis in Laboratory but due to significant advances
and innovation in instrumentation, detector design, data
processing and particle size technology, leads to the
development of Ultra Performance Liquid Chromatography
(UPLC). Principle of UPLC basically remains same only with
the help of technology; it achieves dramatic increases in
resolution, speed and sensitivity of the liquid
chromatography. UPLC Technology is applied throughout the
world for providing, advantage of time, cost and quality.
Beside this it is also reproducible result and robust method
as compared to conventional HPLC.
UPLC has many advantages over conventional HPLC of
which major advantages are speed, quality and cost of
analysis. Due to development of particle size technology and
the use of sub micron particle size in column packing,
significantly reduced in the analysis run time which result in
the development of faster methods and faster analysis of
samples.
((
DETECTORS
1) UV/visible
2) PDA
3) FLR
4) RI
5) MS
We are thankful to Almighty ALLAH for His divine
support.
We are thankful to
Honorable Madam Dr .HUMA ISHAQUE SHAIKH
for giving such a good platform to enhance our skills.
INSTRUMENTATION
UPLC A NEW TREND IN ANALYSIS
NATIONAL CENTRE OF EXCELLENCE IN
ANALYTICAL CHEMISTRY UNIVERSITY OF SINDH
1.Dadu Mal 2.Zubair Ali Shah 3.Rabia Khalid 4 .Ali Haider 5. Shanker Lal
NCEACUNIVERSITYOFSINDHNCEACUNIVERSITYOFSINDH
NCEACUNIVERSITYOFSINDHNCEACUNIVERSITYOFSINDHNCEACUNIVERSITYOFSINDHNCEACUNIVERSITYOFSINDH
NCEAC UNIVERSITY OF SINDH
NCEACUNIVERSITYOFSINDH
NCEAC UNIVERSITY OF SINDH
Separation in chromatography can be explained by Van Deemter
equation which gives relation of resolving power of
chromatographic column with various flows and Kinetic mass
transfer.
HETP = A + B/u + Cu,
Where, HETP is height equivalent to theoretical plate.
A is eddy diffusion.
B is longitudinal diffusion.
C is mass transfer.
u is linear velocity (flow rate).
NCEACUNIVERSITYOFSINDH
NCEAC UNIVERSITY OF SINDH
NCEACUNIVERSITYOFSINDHNCEACUNIVERSITYOFSINDH
CHARACTERSTI
CS
HPLC UPLC
Particle size 3-5µm Less than 2µm
Max: Back pressure 35-40 Mpa 103.5 Mpa
Column Alltima C18 Acquity UPLC BEH C18
Column Dimension 150 × 3.2 mm 150 × 2.1 mm
Column Temp: 30 oC 65-90 oC
Volume Injection 5µL 2µL
Sample Preparation Simple Tedious
Column Coagulation Doesn’t takes place Takes place
Analysis time More Less
Sensitivity Less Higher
AKNOWLEDGMENT
Pharmaceutical Food Environmental Forensic
REFRENCES
World Journal of Pharmaceutical Research Volume 5, Issue 3, 387-394.
Review Article, ISSN 2277– 7105,
(UPLC): A NEW TREND IN ANALYSIS Dr. Sayyed Hussain and Tabrez Shaikh