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POSTED ON NOVEMBER 4, 2017 BY DR. DEEPAK BHANOT LEAVE A COMMENT

Critical considerations in HPLC

solvent selection
Selection of solvents for use as
a mobile phase in HPLC
analysis is a key component of
method development. It is not
possible to hava a universal
solvent which will meet all
applications and more than
often a combination of solvents
is decided based on the
analysis requirements.
Selection of suitable solvents is
based on their physical
properties and compatibilities Solvent selection for mobile phase preparation requires
careful review of its properties
with the sample and column
stationary phase.

The article discusses some of the essential considerations that play an important role in deciding
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suitability of solvents as mobile phase.
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Cost is an important consideration as HPLC requires superior purity grade solvents and it is
common to see dozens of HPLC systems operating round the clock in large laboratories. This
means consumption of high grade solvents in bulk quantities and therefore cost considerations
play a vital role. TOPICS

AAS (53)
Solubility
Analytical Chemistry (3)
The sample should be completely soluble in the mobile phase. Slightest insolubility will result in
phase separations or suspensions which will contribute to operational problems. Calculations (5)

Chemical (2)
Absorbance Chromatography (9)
Generally the detectors used in HPLC are based on absorbance of light by sample contituents.
Environmental Analysis (2)
The intrinsic absorbance of the mobile phase components in the selected wavelength range
should not interfere with the absorbance of the sample. The mobile phase solvent should ideally Featured (2)
have no absorbance at the wavelength of interest. Free e-courses (2)

Gas Chromatography (55)

Volatility
General Topics (140)
The mobile phase solvents should have low volatility especially for use with light scattering
detectors. Highly volatile solvents can lead to compositional changes in mobile phase Guest Posts (27)

composition over use and storage. This can lead to poor reproducibility of chromatograms. HPLC (60)

ICP Techniques (13)

Viscosity
Jobs (1)
The selected solvents should have low viscosity so that ow through the column does not lead to
development of high back pressures. Lab Bytes (17)

Lab Management (95)

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28/04/2019 Critical considerations in HPLC solvent selection - Lab-Training.com

Inertness Laboratory Insurance (8)

Laboratory Safety (13)

The selected solvents should be inert to sample components, column packing and column
material. Any reactivity with any of these components can lead to formation of precipitates, gases Method Validation (1)
or other reaction products which can upset the system performance. The solvents should not
Microbiology (14)
form separate phase on coming in contact with the sample. In other words there should be
complete miscibility of solvents. News (3)

Paper Chromatography (10)

Water pH Measurement (3)
Water is also an important component of reverse phase mobile phase composition either in
Preformulation Studies (3)
combination with other solvents or as bu er media. Water puri cation systems are commercially
available. The water generated by such systems should be used at the earliest possible so that its Quality System (9)
quality does not degrade over the storage period. Separation Science (11)

Spectroscopy (80)
HPLC technique is highly sensitive to changes in the composition of mobile phase reaching the
detector so it is essential to lter the solvents through 0.45μm lter to remove any solid Stability studies (8)
suspensions. Degassing is also an essential requirement to remove any trace of dissolved air
Training (21)
which can lead to ow restriction or spurious peaks.
Weighing (3)

Filed Under: HPLC

About Dr. Deepak Bhanot

Dr Deepak Bhanot is a seasoned professional having nearly 30 years expertise
beginning from sales and product support of analytical instruments. After
completing his graduation and post graduation from Delhi University and IIT
Delhi he went on to Loughborough University of Technology, UK for doctorate research in
analytical chemistry. His mission is to develop training programs on analytical techniques and
share his experiences with broad spectrum of users ranging from professionals engaged in
analytical development and research as well as young enthusiasts fresh from academics who
wish to embark upon a career in analytical industry.

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