Determination of total polyamines in tumor cells by high-performance capillary zone electrophoresis (HPCZE) with indirect photometric detection

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Abstract
Tumor cells contain much higher levels of common polyamines such as putrescine, spermidine, and spermine than normal cells. Our previous work demonstrated that high-performance capillary zone electrophoresis with indirect photometric detection was suitable for the determination of underivatized free polyamines in tumor cells. We now report improved sensitivity (7-fold), greater quantitative reproducibility, and quantitation of both bound and free, underivatized polyamines in PC 12 tumor cells. 1,7-Diaminoheptane was used as an internal standard for quantification. Femtomoles of three polyamines and simple cations such as K+ and Na+ were easily separated in less than 10 min. The quantitations for total spermidine and spermine (bound and free) were found to be 2-fold higher than the quantitation for free, unbound polyamines using our previous method. The signal response was linear over 2 orders of magnitude. The method described here provides a fast and simple technique for quantitation of bound, unbound, and total polyamines for use in the study of the biological role of polyamines.

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