High Performance Liquid Phase Separation of Glycosides. I. Reversed Phase Chromatography of Cyanogenic Glycosides with UV and Pulsed Amperometric Detection

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Abstract
High performance liquid chromatography procedures based on reversed phase chromatography (RPC) using microparticulate octadecylsilica columns were introduced for the separation and detection of some representative cyanogenic glycosides and their degradation products. Pulsed amperometric detection (PAD) provided relatively low detection limits (10−5-10−7 M) for the cyanogenic glycosides and permitted the detection of those lacking a chromophore in their structures (e.g., linamarin) which could not be detected in the UV even at low wavelength. In addition, the PAD was a more selective method of detection when compared to UV at 200 nm, a wavelength at which the molar absorptivity and in turn the detection sensitivity were relatively high for the chromophoric cyanogenic glycosides. However, and in the presence of acetonitrile in the eluent, the detector response in PAD was linear in concentration range over 2 to 3 orders of magnitude as opposed to 4 to 5 orders of magnitude in the UV. Finally, RPC proved very useful in monitoring the rate of hydrolysis of mandelonitrile to benzaldehyde, which is a degradation product of the cyanogenic glycosides amygdalin and pranasin. This hydrolysis was found to be a first order reaction.