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Everyone who bought "Problem Solving in HPLC" by Stavros Kromidas will equally benefit from nearly new practical examples for optimization, trouble-shooting, and instrument performance given in this sequel. The author provides guidance for selecting and evaluating methods, intstruments and columns practical help with everyday A unique approach to solving HPLC problems. 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.
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. Get A Copy.
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More Details Other Editions 3. The worst are those which are not detected but which affect the result anyway. Nevertheless, the book does not distinguish between the two types. The readers decide how they classify them. With increasing experience in HPLC it should become easier to avoid the pitfalls.
The third edition could be expanded with new examples and proposals. Many people helped me with examples, hints or ideas on how to improve the text and gures. I want to thank all of them. Special thanks to the publisher who supports the idea of a picture book, not for children but for novices and experts in the analytical laboratory. I hope that the book will be a useful aid in daily laboratory work thanks to intelligible explanations and lucid illustrations. Gallen, August Veronika R.
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- HPLC/UHPLC Troubleshooting!
This book is not an introductory text to HPLC and also not a troubleshooting guide of the kind what shall I do if my instrument does not work?. It does not replace such books but is intended to complement them. Some texts which, according to my personal opinion, are very useful and should therefore be present in the HPLC laboratory are listed on the next page.
Now this book on your desk is a picture book.
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The gures are at least as important as the texts; sometimes more information can be found in them than could be given in the short descriptions. It is possible, and in principle recommended, to study all the pages in sequence from beginning to end. This method guarantees that one learns about errors which are uncommon and unexpected. On the other hand each pair of pages is limited to one topic, linked to other pages by arrows only, and can therefore be studied in isolation.
The index at the end of the book can help you nd the right pages when a problem occurs, although it must be stated once again that quick troubleshooting advice is not usually provided. Many topics may be absent because this is not a textbook, but the matter presented is of utmost relevance in HPLC. Thus the topics discussed should act as reminders and be used for revision.
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Part II lists the pitfalls and sources of error. They are in a logical sequence, as far as this is possible, following the ow path in an HPLC instrument, from the preparation of the mobile phase to data evaluation. The list is somewhat arbitrary, and not all errors are of equal importance with regard to their possible consequences.
It would, however, be dangerous to distinguish between grave and harmless errors. A minute error can cause much damage under special circumstances. Part III gives some hints on what can be done to avoid errors. Again this synopsis is very heterogeneous in character. This does not diminish its value, of course. Incompleteness is an inevitable feature of this book. I am grateful for all hints on other pitfalls and sources of error or on how to avoid them. XIV Introduction. John W. Dolan and Lloyd R.
Paul C. Lloyd R. Snyder, Joseph J. Kirkland and Joseph L. In chromatography, a physical separation method, the components of a mixture are partitioned between two phases.
One of the phases stays in its place and is called the stationary phase, whereas the other moves in a denite direction and is called the mobile phase. According to the type of mobile phase we distinguish between gas chromatogra- phy, supercritical uid chromatography, and liquid chromatography.
The separation is based upon the different partition coefcients of the sample components between the two phases. It is helpful to divide the chromatographic column into small hypothetical units, the socalled theoretical plates. Within each plate a new partition equilibrium is established. The narrower a theoretical plate, the more equilibrium processes can take place within a column of given length and the more demanding the separation problems which can be solved.
The gure shows the separation of two compounds. One of these prefers the mobile phase but also enters the stationary phase. For the other compound the preference is the other way round. Thanks to this large difference in their properties the two types of molecule can easily be separated.
They are transported through the column by the ow of the mobile phase and thereby reach zones where new equilibria are formed again and again. This height depends on the packing quality of the column, on the mass transfer properties of the phases, and on the analytes involved. Plate height is a function of the particle diameter of the stationary phase. For good columns, plate heights are equal to ca. A ne packing, e. The column with the ne packing can therefore be used for more difcult separation problems.
To judge a chromatogram it is necessary to calculate some data which can be easily obtained.
In addition the breakthrough time or dead time, t0 , must be known although it can be a problem to measure it unambiguously. In principle, the rst baseline deviation after injection marks t0. It depends only on the phase system the types of mobile and stationary phase and on the temperature. For HPLC separations should be 1. The plate number is a measure of the separation performance of a column. The equations given here are in principle only valid for symmetrical peaks. From the plate number it is possible to calculate the height, H, of a theoretical plate e.
More Practical Problem Solving in HPLC
The smaller a peak compared with its large neighbor the greater is the resolution necessary to separate them. It is important to realize that the resolution is inuenced by the three parameters. The separation factor has the largest effect. If a separation needs to be improved it is well worth the effort of increasing , although it is impossible to give a general proposal concerning how to do this.
If the plate number is increased, the effect is only by the factor N; if the column length is, e. Increasing the retention factor only has a notable inuence on resolution if k was small to start with. The upper gure presents several pairs of peaks separated with varying resolution.