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Metformin & Glyburide, A Real Life Application
A 3 minute isocratic analysis on TYPE-C HPLC Columns
It is always more useful to chromatographers to read about a real life separation challenge and how it was
solved. Theoretical situations are very helpful to understand the character of phases but real life case studies
are more practical. Here is a real life situation and how the separation challenge was met and optimized.
The pharmaceutical formulation of the highly polar molecule, Metformin (LogP of minus 2.64) and the relatively non
polar molecule Glyburide (LogP 4.79) are of interest to many Generic Pharmaceutical companies that are considering
its analysis. Ideally a simple quick isocratic analysis was desired.
But Metformin is so polar that when examined, its retention on all of the following Type-B HPLC columns was
unsatisfactory: HPS C18, hQ C8, Axis C18, Axis Phenyl, Cogent F5, HPS Cyano or
HPS Amino using Cogent Mini-Scout Strategy were all used. It should be noted that metformin was too
polar to be retained on C8, C18, cyano or amino columns. Metformin was not retained in 100% aqueous, or buffered a
queous. This was a problem.
Then 2 different TYPE-C phases were examined.
Both TYPE-C Silica and Cogent UDC-Cholesterol columns gave a good symmetrical peak shape for Glyburide,
but a slightly tailing peak for Metformin standards when run independantly.
However, Glyburide was a broad but symmetric peak on a TYPE-C Silica column, but a relatively narrow
symmetric peak could be achieved if the acetonitrile concentration was adjusted so as to give the same retention
time that it had on a Cogent UDC-Cholesterol column with the same column dimensions.
Either TYPE-C Silica or TYPE-C based UDC-Cholesterol columns can be utilized when separating
these two compounds, but the latter was the preferred column because of the better peak width and relative
selectivity after trying both of them.
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