Dear Sudhakar,
I think your product is highly aromatic one. Its better to use UV detector with the wavelength obtained from UV SPEC( its absoption maxima). then use combination of mobile phase either with acetonitrile or methanol. or combination of both. you have to optimize the method. Make the flowrate as 1ml/min. its better to go for isocratic elution rather than the gradient one. Get the standard compound and run with the operating temperature nearly 35degree C. run with different concentration and get the standard graph.
Note: choose the mobile phase based on the solubility of the sample.
Hi Sudhakar,
I too agreed with Ramadoss, if optimized method of HPLC for analysis of (Z)-ethyl 3-hydroxy-2-phenyl acrylate is not available from literature then we can develop method by referring literature of HPLC method of similar compounds and using physicochemical properties of (Z)-ethyl 3-hydroxy-2-phenyl acrylate. I would suggest to develope method by using ODS column , UV detector and Acetonitrile & water as mobile phase for preliminary run. regarding lamda max either use pda detector or scan sample solution of pure (Z)-ethyl 3-hydroxy-2-phenylacrylate in UV-visible spectrophotometer. If you can share some phycochemical properties (Like solubility, melting/boiling point, lamda max, molecular weight, polarity , PKa value etc) of compound then i can suggest some tentative method which can work for such compound.
Regards
Santosh
Dr. Santosh,
This compound is physically liqid and highly soluble in methanol.
Actually we developed one method but compound shows two couple peaks because this compound is contain aldo-enol tautomers.
Finally one thing how to avoid this splitting, i need single peak.
Regards
Sudhakar
Hello Sudhakar,
Could you please let me know the method you are using at present for aldo-enol tautomers, it may be possible to analyse such samples either by addition of some additives in mobile phase like triethylamine or we have to analyse isomeric analytes using normal phase mode.
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