Standardization in the determination of red blood cell polyamines by high-performance liquid chromatography

The choice of the standardization method in the high-performance liquid chromatographic determination of dansyl polyamines (spermidine and spermine) in red blood cell extracts is discussed. 1,6-Hexanediamine, commonly used as an internal standard, is unsuitable for the quantification of spermidine and spermine in red blood cells because their percentage recoveries are significantly different (100% for 1,6-hexanediamine, and 70% for spermidine and spermine). The external standard method and the standard addition method are better suited. The procedure for the preparation of the standard mixture before dansylation has an influence on the values of red blood cell polyamines. Two procedures are compared and the corresponding percentages of variation were found to be high for spermidine and spermine. Thus the procedure in which the standard is treated in a strictly similar way as the red blood cells is certainly the most appropriate one for the quantification.