Conclusion If you choose unfavorable threshold and attenuation, you can always change them later; the raw data are saved. In each case the problem, the solution and the conclusions are presented over a maximum of 4 pages, and in addition the book contains manufacturers’ addresses, references, data tables and checklists. The Case Let us assume that you found the best separation for your particular sample using a C I8 column at pH 9. The solution is to focus according to the manufacturer’s manual; often you only have to tum some adjusting screws to optimize the lamp energy. Comparison of acetonitrile and methanol. The choice of the stationary material does not matter – you could even use ! In some software programs, the peak width is linked to the data rate.

There was a problem filtering reviews right now. The remarkable thing about the new technique was the high pressure to avoid confusion we should speak about high column pressure that gave the name for this technique. In Lieu of an Introduction. The advent of HPLC has not only brought us elaborate instrumentation, but has also made reversed-phase chromatography the leading modality of analytical liquid chromatography. If everything is OK, the peak area is constant; if the peak increases, a part of your sample is sticking to the injection system. Which tests are suitable?

The Solution Preliminary remark If you do not feel competent in the English language, ask your software supplier about manuals in your mother krokidas. This is a direct consequence of the growing importance of certain separation mechanisms.

stavros kromidas more practical problem solving in hplc

You should have flexible software with further options regarding kromdias general data processing: Schaffer, Grunstadt Cover Design: Pump precision or pump constancy We differentiate between ptactical and short-term constancy a Long-term constancy One way to access this solvinv the precision often incorrectly: It’s a first-aid kit for every HPLC user.

  M101P HOMEWORK 5.1

Ln the dead time till is constant and the retention times increases, this means that the pump is OK and the cause is temperature, column or mobile phase.

But let us get back to our original question: Conclusion Please measure the dead volume of your isocratic equipment. Is the chromatogram after the second injection identical to the first one? You have performed a kind of displacement analysis with a miniaturized rising step. We found immense differences see Fig. Remove column, use end fittings to avoid drying and record the conditions in a column log book. He is author and co-author of several articles and numerous successful books.

If you have any deviation, you will have to check the status of the HPLC modules.

Practical Problem Solving in HPLC

If the resulting chromatograms are identical with a reference chromatogram which you obtained with the equipment in perfect condition – maybe after a scheduled master check – there is no need for further checks.

Is the column still in the system or has it been removed? The left hand chromatogram was recorded with a peak width of 0.

stavros kromidas more practical problem solving in hplc

C 18mobile phase: In isocratic elution, flushing is used at the end of a measurement series, to change to another method or merely when necessary, and is also advisable for economic reasons. Also replace the material of the seals. The sample is dissolved in a stronger eluent than the mobile phase e.


Among others some general criteria. Very often, only a defined and constant pH therefore a buffer will allow the desired selectivity and reproducibility. The situation is quite complicated, but the above-mentioned two factors are the reasons in simplified form.

Fewcr problems therefore arise during the separation of polar groups, because no additional interactiom of a polar nature are possible. It is simple to check for a memory effect: The author provides – guidance for selecting and evaluating methods, intstruments and columns, – practical help with everyday trouble-shooting, – advice for optimizing separations, always explaining the reason why. The question now soving How do I prepare a mobile phase?

More Practical Problem Solving in HPLC – Stavros Kromidas – Google Books

There’s a problem loading this menu right now. Of course, the dead volume is more critical the earlier the peaks elute and the more of them there are. The Case Buffered eluents are important for the separation of ionic solutes because they maintain a constant desired pH. The history of chromatography is probably well known, as well as various anecdotes about the subject.

There could be many causes and we only list some of them: The second column contains, for example, 0. What was the problem?

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