Dear all,
I am very new to chiral HPLC.
Can someone help me understand a thing regarding chiral HPLC :
I am studying stereoselective reduction of bulky ketones by recombinant aldo-keto reductase.
This is the reference article : Article Gene mining-based identification of aldo–keto reductases for...
Attaching the same article as well.
So, in this article authors have shown optical purity of chiral alcohols formed by reacting with the recombinant aldoketoreductases.
If I inject +/- or DL or RS 'titled' specific chemical substrates into chiral HPLC column (following standardized protocol) , shouldn't I except resolved peaks like one each for D and L ?
or I must purchase pure R and S forms of all substrates and expect single corresponding peaks ?
If aldoketoreductase converts one of these substrates into either of orientation, will the chiral column be able to resolve such product peak?
So this way, if retention times of standard compound (D /L forms) match with the peak obtained after enzymatic reaction, they could be matched and optical purity could also be estimated in that case...
It is expensive affair to purchase so many pure substrates, so I needed an expert opinion before going ahead! Apologies for so many questions although I tried to search all questions related to chiral HPLC on researchgate.
Sincerely,
Aniket
Dear Aniket Limaye yes. For some general information about racemic mixtures please have a look at the respective Wikipedia entry
Racemic mixture
https://en.wikipedia.org/wiki/Racemic_mixture
Dear Dr. Frank T. Edelmann , thanks a lot.
I understood important convention of using (±) and (+)/(−)
In my case it seems the chemicals are not having 1:1 ratio, having prefix (+)/(−)
So, in order to get the distinct resolutions of the 2 racemic forms ( that is 2 distinct retention times) , would that be possible in case of such chemicals without having 1:1 mixture ?
If its a racemic mixture, I expect there must be 2 clear peaks, isn't it?
Thanks
Dear Zohreh Nazarian
thanks a lot for your detailed explanation.Yes, in fact I have compounds with unequal proportions of R and S forms.
But the good part is I have references giving retention times which could give primary idea about expected retention times for these forms.
Thanks for our help.
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