Hi Mahesh, unfortunately, we do not have a good column for the analysis. Probably the ion-exchange column would work well. Here is a link to a recent app note for pharmaceutical analysis which includes ibuprofen.
Here is a link to our different applications. You can find which column works best depending on which compound you are interested in. Which column is best suitable for separating small oligosaccharides and sugar molecules? C18, C8 or C4? For maltooligosaccharides you can use the following columns in reverse phase. However, anomers will be separated too:. Nicely explained….
I appreciate it… I too have some doubt from the above description, of it is so what is the difference between C18 and H Column or Chiral Colmns… Which column is recommended for Lactic acid Detection and Estimation?
Chiral columns are used to separate different stereoisomers of the same compound. Unlike the linear alkanes used in C18 columns, the stationary phase of chiral columns is usually one stereoisomer of a polymer such as an oligosaccharide. Thank you for this information. I have been using HPLC for normal phase for a while but presently I need to use a reverse phase for purification of protein.
My worry is if I can use the same machine for a reverse phase? If yes, how can I collect a purified sample from the process? The whole thing is quite confusing! This must be done because the solvents used for normal phase chromatography are not miscible with the water and buffers used during reversed-phase.
These flushes should be done with the column removed. The manufacturer of your HPLC may have further guidance. Your email address will not be published. Notify me of follow-up comments by email. Notify me of new posts by email. What Is C18? HPLC has three steps: 1. But what features are the most important? Base Packing Material Silica gel is by far the most common base packing material. Pore and Particle Size The pore size you want is based on the molecular weight of your compound.
Column Dimensions Again, the column dimension used to be standardized along with particle size 4. What is C18?
Kishore on January 23, at AM. Thanks for valuable info. Admin on January 24, at PM. Hi Kishore, Thank you for your comment. Regards, Develosil US Team.
Lingarao on May 6, at PM. Thanks for valuable information Reply. Admin on May 8, at AM. Michael Churchill on September 2, at AM. Hi Anuja, Thank you for reaching out. Shinji Azuma on September 23, at PM. Related: Determination of Theoretical Plates in Columns. Porous stationary phase in these columns allows the separation of the components according to their size.
Combination of polymers like polysaccharides and silica is used as stationary phase in these columns. Small sample molecules penetrate in the pores of stationary phase while the large molecules penetrate partially into the pores. Therefore the large molecules of the sample elute first than the small molecules and this chromatography is called Size Exclusion Chromatography. These columns are generally not used in the analysis of pharmaceutical compounds.
These factors affect the analysis of sample, therefore, these are considered important during the HPLC analytical method development. Columns are selected according to the nature of the compound to be analyzed and the mobile phase. Column performance should also be evaluated time to time generally after runs or as required. Pin it. Ankur Choudhary is India's first professional pharmaceutical blogger, author and founder of Pharmaceutical Guidelines, a widely-read pharmaceutical blog since Sign-up for the free email updates for your daily dose of pharmaceutical tips.
Visitors are also reading:. You can ask questions related to this post here. Setiadjit 05 January. Unknown 10 September. Unknown 28 May. Unknown 04 July. Unknown 17 August. Unknown 24 July. Ankur Choudhary 20 March. The retention times are measured against known standards, thus, the molecules in the sample can be identified.
The longer the molecules spend in the column, the wider and less sharp the peaks become on the chromatogram. The internal pressures on the mobile phase can also reach up to atmospheres.
Normal phase HPLC uses non-polar solvents as the mobile phase and silica particles as the stationary phase. In normal HPLC, polar compounds will stick to the polar silicone longer in the stationary phase compared to non-polar compounds. Therefore, non-polar compounds elute faster in normal HPLC. The pore sizes are generally around 3 microns. In reverse phase HPLC, the stationary phase is modified by hydrocarbon chains typically carbons long , causing the column to become non-polar.
A polar solvent is also used.
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