An Improved High Performance Liquid Chromatographic Method for Identification and Quantization of Polyamines as Benzoylated Derivatives — Oak Academic Publishing
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An Improved High Performance Liquid Chromatographic Method for Identification and Quantization of Polyamines as Benzoylated Derivatives
A simple reversed phase HPLC method was developed for the determination of Putrescine, Cadaverine and Spermidine (a class of polyamines) in their benzoylated form from external known standards. In the optimization procedure, a number of parameters were examined: 1) Solvent used in the extraction of standard polyamines (diethyl ether versus chloroform); 2) Solvent used in the elution of the polyamine (methanol versus acetonitrile); 3) Mode of derivatization and extraction step(s) (derivatization and extraction performed together versus derivatization and extraction performed separately); and 4) Other instrumental parameters (such as UV detection wavelength, gradient profiles). The advantages of our method, relative to the standard Morgan method are: a) decreased chromatographic runtime, b) ease of preparation with good resolution, sensitivity, and reproducibility using a standard RP-HPLC method.
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