Original scientific paper

https://doi.org/10.5599/admet.3.4.249

Rapid identification of bioactive carbohydrazide reaction products by an LC-DAD-SPE-NMR approach

Iva Habinovec ; University of Zagreb, Faculty of Science, Department of Chemistry, Horvatovac 102a, 10000 Zagreb, Croatia
Tomislav Jednačak ; University of Zagreb, Faculty of Science, Department of Chemistry, Horvatovac 102a, 10000 Zagreb, Croatia
Predrag Novak ; University of Zagreb, Faculty of Science, Department of Chemistry, Horvatovac 102a, 10000 Zagreb, Croatia

Abstract

On-line coupling of high performance liquid chromatography with diode array detection to solid phase extraction combined with nuclear magnetic resonance (LC-DAD-SPE-NMR) was used to monitor carbohydrazide condensation reaction progress. First, a chromatographic method was developed and optimised and individual peak separation was readily achieved by using an isocratic acetonitrile-phosphate buffer mobile phase. Subsequently, separated compounds were trapped on SPE cartridges and dried with nitrogen gas. Peak elution was then performed with deuterated acetonitrile and sent for NMR analysis. Single and multiple trapping options were applied. One- and two-dimensional NMR spectra were recorded using a Prodigy cryoprobe. It was demonstrated that LC-DAD-SPE-NMR setup was proved very useful for rapid and unambiguous identification of the reaction products and for determination of their structure. By using Prodigy cryoprobe in NMR measurements we were able to detect and identify compounds present at microgram level thus proving a high sensitivity of this methodology for monitoring reactions of bioactive molecules and drugs.

Keywords

carbohydrazide derivatives; reaction monitoring; LC-DAD-SPE-NMR; product identification; structure elucidation

Hrčak ID:

150559

URI

https://hrcak.srce.hr/150559

Publication date:

30.12.2015.

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