Fluorometric assay for polyamines in urine and tissues using electrophoresis on Titan III cellulose acetate.
A simple, rapid assay method for polyamines (putrescine, spermidine, and spermine) in urine and tissues using electrophoresis on Titan III cellulose acetate was developed. In this procedure, polyamines are preliminarily extracted from a hydrolysate of urine or from supernatants of tissue homogenates by use of a Bio-Rex 70 minicolumn. After electrophoretic separation, polyamines are fluorometrically detected by the reaction with o-phthalaldehyde and 2-mercaptoethanol. Six extracts and two external standards of polyamines can be separated and detected in 11 min on a cellulose acetate strip. This method permits the determination of polyamines in a range of 0.1 mM (25 pmol) to 1.0 mM (250 pmol).
Kanda
S
S
Department of Chemistry, Sasaki Institute, Chiyoda-ku, Japan.
Takahashi
M
M
Nagase
S
S
D000818:Animals; D002482:Cellulose / Q000031:analogs & derivatives*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004586:Electrophoresis / Q000379:methods; D005470:Fluorometry / Q000379:methods; D006863:Hydrogen-Ion Concentration; D008623:Mercaptoethanol; D011073:Polyamines / Q000032:analysis / Q000652:urine*; D051381:Rats; D012015:Reference Standards; D001749:Urinary Bladder Neoplasms / Q000652:urine; D009764:o-Phthalaldehyde
D011073:Polyamines; C005062:acetylcellulose; D008623:Mercaptoethanol; D009764:o-Phthalaldehyde; D002482:Cellulose