UPLC uses smaller particle sizes (<2 microns) than HPLC (3-5 microns) which allows it to operate at higher pressures and provide faster, more sensitive and selective separations compared to HPLC. Some key differences are that UPLC uses smaller diameter columns, achieves higher resolution and plate counts, reduces run times, solvent consumption and cost of operation compared to HPLC. However, the smaller particles used in UPLC columns have a narrower usable lifespan than HPLC columns.

1. DIFFERENCE BETWEEN HPLC
AND UPLC
Dr. Vrushali Sachinkumar Tambe
Professor (Department of Pharmaceutical Chemistry)
PES Modern College of Pharmacy (for Ladies), Pune,
India

2. • High-performance liquid chromatography (HPLC) is an
important liquid chromatography (LC) technique used
for the segregation of different components in
mixtures. It is also used for the identification and
quantification of compounds.
• In 2004, Waters launched and trademarked Ultra
Performance Liquid Chromatography (UPLC) which is
based upon small, porous particles (sub 2micron
particles). This technique achieved dramatic increase in
resolution, speed and sensitivity in LC.

3. Sr. No. UPLC HPLC
1 Particle size of stationary phase
is less than 2 micron
(typically 1.7 micron)
Particle size of stationary phase is
between 3 to 5 micron
2 Operates at higher pressure due
to reduced particle size
Maximum backpressure of 1000
bars/ 100 MPa
Operates at relatively lower
pressure than UPLC
Maximum backpressure between
300-400 bars
3 Inner diameter of the column is
generally 0.75-1.8 mm
Inner diameter of the column is
3-10 mm

4. Sr. No. UPLC HPLC
4 Dimension of column
150* 2.1 mm or 1mm
(length* id)
2-15 cm length
Dimension of column
150* 3.2 or 4.6 mm
(length* id)
2-15 cm length
5 More selective and sensitive Less selective and sensitive
6 High resolving power Less resolving power

5. Sr. No. UPLC HPLC
7 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
8 It decreases the consumption of
solvent and increases sample
throughput
More solvent consumption,
less sample throughput
9 The higher back pressures compared
to conventional HPLC decreases the
life of the columns (column
coagulation). Increasing the column
temperature reduces the back pressure
problem in UPLC.
Less back pressure, more
life of column

6. Sr. No. UPLC HPLC
10 The particles of less than 2 μm are
mostly non-regenerable and,
therefore, have a narrow use.
Has more use
11 Higher precision in sample
introduction is required
Lower comparative to UPLC
12 Detectors uses small flow rates
and low detection volume, Hence
high data sampling rate is
required.
Detectors uses higher flow rates
and high detector volume

7. Sr. No. UPLC HPLC
13 Stationary phase is modified to
withstand high pressures. HYBRID
of inorganic silica and organic group
is used.
Bridged ethylene hybrid (BEH),
High strength silica (HSS), Charged
surface hybride (CSH) are used as
stationary phase.
Inorganic Silica modified to
different polarity is generally
used.
14 Injection volume are small
(generally 2 microliter)
Injection volume are large
(generally 5 microliter)
15 Less run time More run time

8. Sr. No. UPLC HPLC
16 More plate count is achieved
during analysis
Less plate count is achieved
during analysis
17 System should withstand high
pressures, more efficient pumps
are required
Less efficient pumps are
required
18 Low extra column dispersion More extra column band
broadening