28/04/2019 How to ensure trouble-free HPLC System Operation

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POSTED ON FEBRUARY 11, 2014 BY DR. DEEPAK BHANOT LEAVE A COMMENT

How to ensure trouble-free HPLC

System Operation
HPLC is the major support system in
laboratories dealing with separation and
identi cation of  complex organic
compounds. In industrial sector some
laboratories are equipped with several
HPLC systems in Quality control and R&D
sections which run like workhorses 24 X7
hours for 365 days in a  year. Downtime
of these systems proves to be very
expensive in terms of production and
loss of man-hours so It becomes
extremely important to keep HPLC HPLC Preventive Maintenance
systems up and running.

The HPLC system comprises of di erent

modules and things can go wrong unexpectedly. It becomes essential to follow preventive
maintenance schedules religiously even when the system is giving excellent performance. Many
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comprehensive HPLC texts are available which deal with pages and pages on the subject of
preventive maintenance but the objective of this short article is to provide some valuable tips TODAY WEEK MONTH ALL
that can ensure trouble-free system operation.
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Mobile phase
Mobile phase is the lifeline of your HPLC. Any disruptions in the ow can result in unstable
baseline and erratic uctuations in retention time of peaks. TOPICS

Use high purity grade solvents and HPLC grade water for preparing mobile phase mixtures AAS (53)

Mobile phase should be prepared fresh every time. Analytical Chemistry (3)
It is advisable to lter even HPLC grade mobile phase solvents prior to use under vacuum
Calculations (5)
using 0.45 µ lter
Clean or replace online lters regularly as a practice Chemical (2)
Degas the mobile phase on preparation even if your system has inbuilt degassing facility.
Chromatography (9)
When mobile phase requires to be changed ush the system with the solvent having
intermediate polarity and then only run on the new mobile phase Environmental Analysis (2)

Flush the system with at least 200 ml of water after using bu ered mobile phases. Bu ers if Featured (2)
allowed to dry out lead to formation of solid crystalline deposits which can damage pistons
Free e-courses (2)
and plunger seals.
Gas Chromatography (55)

Sample General Topics (140)

Try dissolving sample in mobile phase. If this is not possible then try dissolving in a solvent Guest Posts (27)
of close polarity
HPLC (60)
Filter sample prior to injection
Rinse sample vials with sample solution before loading onto the autosampler. ICP Techniques (13)

Jobs (1)
Column Care
Lab Bytes (17)
Column is a critical component and needs proper attention and care :
Lab Management (95)

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28/04/2019 How to ensure trouble-free HPLC System Operation

Avoid damage to packing by exposure to thermal or mechanical shocks Laboratory Insurance (8)
Avoid sudden change of operating conditions such as ow rate and pressure
Laboratory Safety (13)
Use guard columns ahead of the main column
Before storage purge out bu ers or mobile phase. For long storage purge and store with Method Validation (1)

the manufacturer recommended solvent and close the end caps Microbiology (14)
Use columns within the pH range 2 – 8.
News (3)
It is a good idea to keep a stock of frequently used columns
Paper Chromatography (10)

Pump care pH Measurement (3)

The e ciency of your system will depend on the capability of the pump to always give Preformulation Studies (3)
consistency of ow rates.
Quality System (9)

Maintain stock of recommended essential spares such as o-rings, plungers, check valves, etc Separation Science (11)
Operate well within the speci ed pressure limits to prevent damage to the component parts
Spectroscopy (80)
Do not allow bu ers to dry inside the pump
Clean seals and o-rings periodically as per supplier recommendations Stability studies (8)

Clean online lters at regular intervals Training (21)

Prime pump with new mobile phase to remove previous mobile phase components
Weighing (3)

Injector care
In manual injection rinse sample loop with sample to prevent memory e ects
Flush with iso- propanol in between injections and also when using the auto sampler

Detector care
Detector response decides the nal output of your results. Care and maintenance of detector has
same importance as any other HPLC component :

With continuous use the lamp can contribute to baseline noise and drift. Keep in stock a
spare lamp and replace as per supplier recommendations
User HPLC grade solvents for mobile phase preparation. Always degas solvents to prevent
bubble formation inside the detector cells
Clean cell o -line when required with 10% nitric acid or chromic acid. Flush with several
volumes of water and nally with the mobile phase
Do not exceed the prescribed pressure limits to prevent damage to the cell walls

The tips provided can ensure long trouble-free operation of the HPLC system. We shall
appreciate receiving comments based on your experience on this important topic.

Filed Under: HPLC Tagged With: High-performance Liquid Chromatography, Hplc, Hplc Grade Water,
Hplc System

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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