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Pure Practical Series




The Pure Practical Series is a set of one-day courses which contain only hands on HPLC experience, run in the labs at Laserchrom HPLC Laboratories Ltd in Rochester, Kent, UK.

For this year there are five possible practicals to choose from. Participants will work alone or in pairs, and all practicals run together on the same day. We have four Pure Practical days scheduled in 2012 - see the Course Calendar.


There are four different methods for calculating Limit of Detection and Limit of Quantitation. These are described in the ICH Guidelines, the Pharmacopoeas and in Lloyd Snyder's Introduction to Modern Liquid Chromatography. Problem is that each tends to give a different answer!

During the course you will use each method and compare the results!

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2: Fast HPLC using new fused core particle columns

3: Fluorescence detection with post column derivatisation

A number of new fused core packings have become available, and these allow the possibility to achieve results close to uHPLC from a standard HPLC instrument.

This course allows you to try the new columns and compare the results to those from a standard HPLC column. Prepare to be impressed!

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During this course you run samples on a gradient HPLC system using a fluorwescence detector and psot-column detection. You will learn how to set up such a system, and how to use it without breaking the fluorescence flow cell!

Fluorescence is a very powerful tool for sensitivity in HPLC, but samples often require derivatisation. During this course we use OPA.

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4: HPLC with Diode Array Detection

5: Consistent Results with Area % calibration


Diode array detection offers a dramatic extension to the capabilities of an HPLC system without a huge additional cost.

We use primarily an Agilent 1100 DAD, and show the difference in spectra at 1024 diodes and 256 diodes. During the course you will see what a solvent front and UV cut-off look like with DAD, and learn to use the detector settings to good advantage. We also look at factors which change the observed UV spectrum, and which could therefore affect peak identification

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We get more queries from Area% calibrations than anything else! So this course shows how to set it up, what sample concentration to use, how to get a reproducible gradient blank, and how to get accurate and consistent results

During the course you will run two samples, one pure and one less pure, and learn how to identify which peaks are impurities and include them in the Area% calculation.

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