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

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.