Connecting People, Science and Regulation ®

Ultra High Performance Liquid Chromatography (UHPLC):

Opportunities and Challenges for the
Quality Operations Laboratory

Jack Leary, PhD

Global Quality Operations
Pfizer Global Manufacturing
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Why is the implementation of new technology important for

Pharmaceutical Q
Qualityy Operations
p laboratories?

Productivity improvements accelerate speed of lab

decisions, which translates to less idle time for
manufacturing equipment
 Connecting People, Science and Regulation ®

Why is the implementation of new technology important for

Pharmaceutical Q
Qualityy Operations
p laboratories?

Productivity improvements accelerate speed of lab

decisions, which translates to less idle time for
manufacturing equipment

For the phamaceutical industry, how is new technology

defined in the analytical
y and regulatory
g y arena?

Depends on frame of reference

 Connecting People, Science and Regulation ®

Why is the implementation of new technology important for

Pharmaceutical Q
Qualityy Operations
p laboratories?

Productivity improvements accelerate speed of lab

decisions, which translates to less idle time for
manufacturing equipment

For the phamaceutical industry, how is new technology

defined in the analytical
y and regulatory
g y arena?

Depends on frame of reference

One example of new technology is Ultra High Performance

Liquid Chromatography
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What is UHPLC?

Ultra High Performance Liquid Chromatography (UHPLC)

uses:

• Chromatographic columns with smaller diameter particles

(<2um)

• System
y pressures which are ggreater than typical
p yp High
g
Performance Liquid Chromatography (HPLC), e.g. > 6000
psi
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A Technique with Many Names:

•Ultra High Performance Liquid Chromatography (UHPLC)

•Ultra High Pressure Liquid Chromatography (UHPLC)
•High Speed Liquid Chromatography
•Fast Liquid Chromatography

Ultra Performance Liquid Chromatography (UPLC) – Waters

Rapid Resolution LC System (RRLC) – Agilent
X
Xtreme Li
Liquid
id Chromatography
Ch h (X
(X-LC)
LC) – Jasco
J
Accela High Speed Chromatography System - Thermo
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Effects and Potential Benefits of sub 2 um particle size and

higher
g column ppressures:

•Decreased run time

•Improved sensitivity
•Enhanced peak resolution
•Decreased solvent consumption
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Development of UHPLC Method or Conversion of Existing

HPLC Method

UHPLC is based upon the same principles of HPLC.

However, since the column packing material is of smaller
diameter, modification of the chromatographic conditions
and/or sample preparation are often needed.
needed
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Development of UHPLC Method or Conversion of Existing

HPLC Method

When converting an HPLC method to UHPLC, use of the

same type of column/packing material is recommended, when
possible, with the exception of the particle diameter and the
column dimensions
dimensions. It is also advisable to filter mobile phase
with 0.20 um filter, use a titanium “stone”, and install
precolumn filters to minimize column blockage
p g from
particulates.
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UHPLC Flow Rate

The fl
Th flow rate off UHPLC methods
h d generally
ll range from
f 0.25
0 25 –
0.75 mL/min, but are dependent upon the column
backpressure.
backpressure

Focus on system
y p
pressure is important.
p Currently,
y Agilent
g
RRLCs can only be used up to a maximum of ~9000 psi while
Waters UPLCs can be used up to 15000 psi. Therefore, it is
i
important
t t tot consider
id the
th type
t off iinstrumentation
t t ti that
th t will
ill be
b
used at all the site(s) running the UHPLC method being
developed before selecting a final flow rate for the converted
method.
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UHPLC Injection Volume:

Typical injection volumes range from 0.3 – 0.8 µL, but is

dependent upon peak shape and peak intensity.

Although injection volumes for UHPLC are less than for

HPLC, improvements in sensitivity are often realized. The
reason for the improved sensitivity is because of the greater
efficiency and tighter eluting bands.
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Other UHPLC Recommendations:

•The dead volume from the column outlet should be kept to a

minimum to reduce band broadening effect

•Cell volumes of detectors may need to be reduced for the

same reason and split ratios into MS detectors may need to be
reduced or eliminated.
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Sample Gain in Efficiencies

HPLC vs UHPLC

Data from Waters Corp.

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Comparison of HPLC and UHPLC Assay& Deg Method for Capsules

Deg 2 API 9.01min

Run Time-16min
4.72 min
Inertsil, ODS-2, 4.6x150mm,5µ
Flow Rate: 0.6mL/min Deg 1
Inj. Vol: 20µL 3.80min
M.P. 0.05M Potassium Phosphate
pH3.5:ACN(45:55)

Deg 1 0.15 min

Run Time-0.4min API 0.34min
1.7µm C18 2.1x50mm
Flow Rate: 1.5mL/min
Inj. Vol: 0.4µL
M.P. 0.05M Potassium Deg 2 0.25
Phosphate min
pH3.5:ACN(45:55)
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HPLC Chromatogram for Cleaning Method (22 min)

UHPLC Chromatogram for Cleaning Method (4 minutes)

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Industry Considerations When Implementing New Technology

• Requirements for comparison of new technology to existing
technology, including, where applicable assuring equivalence with
compendial methods
ƒ Mode of regulatory change will be driven by determination whether
the new technology an extension of your old technology, brand new
t h l
technology for
f thi
this application?
li ti ? UHPLC is i an extension
t i off existing
i ti
technology (HPLC)
ƒ Weight and extent of method validation is also linked to whether
this is a new technology or an extension of existing technology
ƒ UHPLC equipment
q p are relatively
y new models: Need to determine
robustness and reproducibility for long term routine testing; New
technologies have a small customer base and experience has not
been developed.
developed
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Strategies for Implementing New Technology

• Focus on areas of operation where mode of regulatory change

is self-approvable, such as HPLC to UHPLC conversion for
Cleaning Verification
• Allows Control Labs to gain familiarity with instrumentation
• Interface the creation of the new technology in tandem with
the ongoing with development and process improvements of
manufacturing process improvements
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Pfizer Experience with UHPLC

• Many of our manufacturing site based laboratories have either

purchased or have plans to purchase UHPLC systems

• Waters & Agilent systems have been more commonly acquired

• Local vendor support for introduction of this new technology

has been productive

• Current focus has been on self approvable new technology

extensions
i
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Acknowledgements

Jeff Snider
Edgardo
g Ortiz
Leon Clark,
Scott Afman,
Avish Ramcharan
Prasad KPP