28/04/2019 How are analytical scale and preparative scale HPLC different?

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POSTED ON AUGUST 22, 2017 BY DR. DEEPAK BHANOT LEAVE A COMMENT

How are analytical scale and

preparative scale HPLC di erent?
HPLC is a versatile analysis tool that a ords
separation and quanti cation of large range
of chemical compounds. It nds unlimited
applications in basic research,
pharmaceuticals, foods, polymers and other
similar applications. The availability of
numerous columns, stationary phases,
mobile phases and detectors makes HPLC a
high utility facility in modern laboratories.

Analytical scale HPLC is the building block

Preparative HPLC system (Image Courtesy :
on which the edi ce of other HPLC http://www.nis.unito.it)
techniques such as ion exchange
chromatography, gel chromatography and
preparative chromatography stand.
Developments in these related branches would not have been possible without preliminary
studies carried out on analytical HPLC. The objective of the present article is to o er clarity on
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how analytical HPLC is di erent from preparative HPLC.
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The rst and foremost thing to keep in mind is that the basic principles are common to all the Jetpack plugin with Stats module
mentioned HPLC techniques, e.g., sample introduction, separation of sample components in the needs to be enabled.
column and nally detection using a range of detection techniques. To develop a suitable
separation using preparative separation you have to rst carry out preliminary studies using
analytical HPLC and suitably upgrade to preparative scale. The basic di erence between the two
techniques is the scale of operation which will be highlighted along with other di erences below: TOPICS

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Clarity on the Objective of Analysis
Analytical Chemistry (3)
Before you proceed with the analysis you should have a clear understanding of objective of
Calculations (5)
analysis. Do you wish to have information regarding nature of components of sample followed
by their quantitative estimation or you wish to separate components of interest followed by their Chemical (2)
collection in desired quantities for future reference and use. In the rst case should you have to
Chromatography (9)
use analytical scale separation whereas in the latter you have to resort to collection of the
separated components. Environmental Analysis (2)

Featured (2)
Scale of Operation Free e-courses (2)
In analytical scale separations the quantity of samples injected is small, usually in micro liter
Gas Chromatography (55)
levels whereas in preparative scale separations injection volumes are large so as to yield pure
compounds in milligrams to gram levels. The column eluent instead of being led to a waste General Topics (140)

container is led into separate containers in a fraction collector after passing through a non- Guest Posts (27)
destructive detector. Automated switching valve in the system permits automatic collection of
HPLC (60)
fractions in di erent receptacles depending on the peak response of the detector. Multiple
injections can be made to purify and collect larger quantities. ICP Techniques (13)

Jobs (1)
Column Dimensions
Lab Bytes (17)
Preparative HPLC columns are large in comparison to analytical columns. The analytical scale
Lab Management (95)
columns have diameter of 4.6mm whereas preparative column id’s are generally in the range of

lab-training.com/2017/08/22/analytical-scale-preparative-scale-hplc-different/ 1/3
28/04/2019 How are analytical scale and preparative scale HPLC different?

50 – 200 mm. Laboratory Insurance (8)

Laboratory Safety (13)

Analytical columns are packed with stationary phase particle sizes ranging between 3-5μm in
Method Validation (1)
comparison to preparative scale columns which are packed with larger particles above 10μm
size. Such particle sizes permit higher ow rates in columns. The longer length and wider column Microbiology (14)
diameters o er lower back pressures.
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Injection Mode Paper Chromatography (10)

pH Measurement (3)
Analytical level separations generally involve large number of samples along with small sample
injection volumes so an autosampler is the preferred mode of injection. Conversely, in the Preformulation Studies (3)
preparative scale separations the number of samples is small or there could be even a single
Quality System (9)
sample y but injection volumes are larger so large volume injection loops are used.
Separation Science (11)

Choice of Detectors Spectroscopy (80)

Due to the analysis requirements only non-destructive detectors such as UV, uorescence, or Stability studies (8)
refractive index detectors are commonly used for preparative scale applications. However, due to
Training (21)
large sample volumes the detectors have larger volume ow- cells in comparison to analytical
scale separations. Weighing (3)

To summarize some of the operational parameters are mentioned in the table below :

Parameter Analytical HPLC  Preparative HPLC

L=150cm or more, Id=50-
Column dimensions L=100-250mm ,id= 4.6mm
200mm
Particle size 3-5μm 20μm-50μm

Flow rates 0.1-10 ml/min 10-200ml/min

 

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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