Explain how we choice exact column during reverse phase chromatography
This is too broad a topic for a web forum. Please invest your own time to read one of the informative classic texts on the subject of HPLC method development. Many of these texts include some general guidelines for new to intermediate level users. Here is the one that I most commonly recommend to my students and clients.
- "Practical HPLC Method Development"; Snyder, Kirkland and Glajch. 2nd edition (1997). ISBN 0-471-00703-X. (*which includes a section with initial column selection advice as well as information on how the sample properties and goals of separation direct the choice).
Hiii , The hydrophobic subtraction model evaluates the selectivity of HPLC reversed-phase columns so that researchers can choose a suitable substitute or a sufficiently orthogonal second column ....there are certain factors for column selection , one of them is selectivity factor , capacity factor ...which depends on this .,,,the bonded phase chromatography of certain columns which are mainly used in RPC
hope these article will help u a lot Muhammad Amjad
It is 'really' the molecule of interest (in the sample matrix) that determines the means of retention (the column, the mobile phases, and the detector). Your question is WAY too broad.
Hello, It better to start with your analytes and then come to the column selection.
Please look at the following below links which may help you in your analysis:
https://pubs.acs.org/doi/pdf/10.1021/ac071905z
https://www.obrnutafaza.hr/pdf/tosoh/kolone/TSKgel-RPC.pdf
https://www.sigmaaldrich.com/technical-documents/protocols/biology/hydrophobic-interaction/rpc-purification-options-with-source.html
Thanks
Sasikala M wrote: "c18 ODS column is best for RPC elution" ????
Best? No. Especially when no sample type, or specific application is presented. Is it a common type of RP column? Yes, the "C18" is, but the question asked was HOW TO CHOOSE a RP Column (not name one type of generic RP column). *Advice is only useful when it addresses the question or concern.
Dear William Letter,
What will be the pH of the phosphate buffer during the simultaneous determination of dexamethasone sodium phosph and prednisolone acetate during HPLC analysis on C18 column. Kindly help me.
Thanks and regards
Muhammad: If you have a new, unrelated question, then please delete your duplicate posts and start a new thread.
PS: Please research your own questions first, before posting asking others to do it for you. To obtain suggestions on HPLC methods, please use a search engine (i.e. GOOGLE) with the appropriate search words to find example applications, not this forum.
Dear Muhammad,
Selecting the best column for your application is usually a bit of trial and error but you can narrow down the options tremendously by keeping your application in mind. Your column length, ID, particle size, and phase are important and influenced by your sample as well as instrument limitations. It is also important to keep in mind the solvent system you are using might influence your elution profile (example methanol and acetonitrile can give different elution profiles on a phenyl phase).
Focusing only on the phase, in order to select the best RP column, it is important to know the analytes of interest. You ultimately want to match your compound's functional groups to that of the column you want to use.
Consider if your compound has potential for hydrogen bonding, Pi-Pi interactions, weak ionic, dipole-dipole, or hydrophobic interactions. Then match the columns functional groups to these.
There are a variety of RP columns on the market including a range of differently modified C18 columns (C18 with polar embedded groups, C18 end-capped, non-end capped, C18 with pH stability, 100% aqueous C18, ...) beyond this there are also phenyl columns, cyano columns and amino columns that can all run in RP. Your chromatography supplier should be able to guide you in what columns are available, their functional groups, and what would be recommended for your application. This information can also be found in column manufacturer brochures and on their websites
Example: https://www.phenomenex.com/Kinetex/Selectivities
In some cases, it may be easier to look for an application note on the compound of interest than trying to match a column to your application. Please have a look at the below link that can be of use:
https://www.phenomenex.com/Application/Search
https://chromaappdb.mn-net.com/
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