Estimation of urinary diamines and polyamines by thin-layer chromatography

Area of Science:

• Biochemistry
• Analytical Chemistry
• Clinical Chemistry

Background:

• Polyamines are essential biological molecules implicated in cell growth and proliferation.
• Altered polyamine levels are associated with various diseases, including cancer.
• Accurate quantification of polyamines in biological samples is crucial for diagnostic and research purposes.

Purpose of the Study:

• To develop and validate a thin-layer chromatography (TLC) method for separating and quantifying dansylated polyamine derivatives.
• To investigate polyamine levels in human urine, specifically comparing cancer patients and healthy individuals.

Main Methods:

• Dansylation of polyamines (ammonia, 1,3-diaminopropane, 1,4-diaminobutane, 1,5-diaminopentane, spermidine, histamine, spermine) followed by separation using ascending TLC on silica gel.
• Quantification of specific polyamines (putrescine, cadaverine, spermidine, spermine) in human urine by direct scanning of fluorescent intensity.

Main Results:

• Successful separation of dansylated polyamine derivatives was achieved using a chloroform-triethylamine solvent system.
• The method allowed for the direct quantification of putrescine, cadaverine, spermidine, and spermine in urine samples.
• Significantly higher concentrations of spermidine and spermine were detected in the urine of cancer patients compared to normal urine samples.

Conclusions:

• The developed TLC method provides a reliable approach for the analysis of polyamines in biological fluids.
• Elevated urinary levels of spermidine and spermine may serve as potential biomarkers for cancer detection.