Isolation and determination of tissue spermidine using Bio-Rex 70.
Bio-Rex 70, a weak cation exchange resin, has been used to extract spermidine from rat tissues. This method compares favourably to other extraction procedures with respect to selectivity, reproducibility and time needed to perform the procedure and, because it combines the optimum of these procedures, it appears more adaptable for routine laboratory use. Following extraction, spermidine is quantified fluorometrically after condensation with o-phthaldialdehyde (OPT). The sensitivity of the present procedure allows for the chromatographic isolation of 20 ng spermidine to give fluorescence twice that of blank. The procedure is selective for spermidine because the fluorescence of other OPT-reactive substances has been eliminated by either the extraction or the assay procedures. The stability and reproducibility of the adsorption and elution characteristics of Bio-Rex 70 enables the determination of 30 tissue samples in a working day. This procedure has been used to determine the levels of spermidine in whole and regional brain areas. In addition, the concentration of spermidine in peripheral tissues was also determined. Of the brain regions, the medulla oblongata/pons had the highest level of spermidine (68.24 +/- 7.67 microgram/g), and in peripheral tissues the highest level of this polyamine was found in the heart (48.59 +/- 4.32 microgram/g).
D000818:Animals; D002411:Cation Exchange Resins; D005260:Female; D005453:Fluorescence; D008297:Male; D051381:Rats; D012117:Resins, Synthetic; D013050:Spectrometry, Fluorescence; D013095:Spermidine / Q000302:isolation & purification*; D009764:o-Phthalaldehyde
D002411:Cation Exchange Resins; D012117:Resins, Synthetic; D009764:o-Phthalaldehyde; C026257:Bio-Rex 70; D013095:Spermidine