28/04/2019 Ion pair chromatography in reverse phase HPLC

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POSTED ON NOVEMBER 28, 2013 BY DR. DEEPAK BHANOT 2 COMMENTS

Ion Pair Chromatography – a

viable alternative for Separation of
High Polarity molecules by
Reverse Phase HPLC
Ion pair chromatography helps separate charged analytes using the popular reverse phase
chromatography. Ion pair reagents enhance peak shape and retention time when modifying
mobile phase ratios or changing stationary phase is not of much help.

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Ion Pairing Schematic TOPICS

AAS (53)
The technique was developed by Dr. Gordon Shill in 1973. Ion paired reagent has an ionic Analytical Chemistry (3)
terminal functional group at the end of a non polar chain. The reagent is added to the mobile
phase and given su cient time to equilibrate with the column. The non polar end of the chain is Calculations (5)

held strongly to the non- polar stationary phase whereas the charged end sticks out into the Chemical (2)
mobile phase. Ionic spieces of opposite charge in the mobile phase get immobilized by the
Chromatography (9)
charge on the reagent, resulting in neutral ion pairing between the sample molecule and the
reagent. This helps in selective retention of charge to analytes in reverse phase HPLC. Environmental Analysis (2)

Featured (2)
Ion exchange chromatography had been in existence for several years but introduction of Ion
pair chromatography o ered several advantages: Free e-courses (2)

Gas Chromatography (55)

Simple preparation of bu ers
General Topics (140)
Wide choice of carbon chain lengths for improved retention and separation
Signi cantly reduced separation time Guest Posts (27)
Simultaneous separation of both ionized and non-ionized molecules
HPLC (60)
High reproducibility of results
Improved peak shapes ICP Techniques (13)

Jobs (1)
Analysis of acidic samples is carried out by adding quatrenary ammonium salts such as tetrabutyl
Lab Bytes (17)
ammonium chloride to the mobile phase
Lab Management (95)

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28/04/2019 Ion pair chromatography in reverse phase HPLC

Similarly a negatively charged reagent such as an alkyl sulphonic acid can be used to selectively Laboratory Insurance (8)
retain positively charged ionic bases
Laboratory Safety (13)

Method Validation (1)

Microbiology (14)
Precautions in use of Ion pair reagents :
News (3)

Ion pair reagents selected should lack absorbance in UV or uorescence wavelength region Paper Chromatography (10)
selected for analysis
pH Measurement (3)
Both mobile phase composition and temperature should be controlled precisely so gradient
elution is ruled out when using ion pair reagents Preformulation Studies (3)
Allow su cient time for the mobile phase to equilibrate with the column so that changes in Quality System (9)
mobile phase or temperature are minimized
Separation Science (11)
The column used with ion pair agents should be dedicated for such analysis as it is di cult
to completely ush out ion pair reagents even after extensive ushing Spectroscopy (80)

Stability studies (8)

In case you have experience with the use of Ion pair separations please leave your comments
and share your experiences with others. Training (21)

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.

Comments

Reversing says:
June 11, 2014 at 7:49 pm

Its like you read my mind! You appear to know a lot about this, like you wrote the
book in it or something.

I think that you can do with some pics to

drive the message home a bit, but other than that, this is great blog.
A fantastic read. I will certainly be back.

Reply

Dr. Deepak Bhanot says:

June 13, 2014 at 9:28 am

Your suggestions are always welcome

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28/04/2019 Ion pair chromatography in reverse phase HPLC

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