Polyamines-an improved automated ion-exchange method

An accurate, precise, and improved automated cation-exchange chromatographic method with ninhydrin detection for the analysis of di- and polyamines (putrescine, cadaverine, spermidine, and spermine) has been developed. We have shown that different types of biological fluids such as urine, blood plasma, blood sera, tissue extracts, and cancer cell culture media can be analyzed under identical chromatographic conditions. The simplicity and precision of the method was achieved by eliminating the sample pre-separation and using an internal standard technique. Thus, not only has the sample preparation been simplified, but the accuracy and precision and sensitivity of the method have been greatly improved. Twenty-four unattended analyses were performed each day. With minor modifications of the instrument a two-fold analytical output can be achieved with analysis time cut to 30 min. The ruggedness and applicability of the method has been demonstrated in our laboratory during the past six months. More than two thousand urine and hundreds of other physiological samples have been analyzed by this method with a relative standard deviation from 3.3 to 7.8%, and recoveries of 94 to 97%. This automated ion-exchange chromatographic method for the polyamines will be useful to researchers in biological markers programs for monitoring the course of cancer and effectiveness of chemotherapy.