The TLC based separation and quantification method described here has been successful in quantifying DOX levels in actual clinical patient samples with 75% the DOX content falling within 15% of the HPLCderived values and 100% falling within 16%. This single mobile phase TLC method meets the FDA acceptance criteria for a bioanalytical system using chromatographic methods . This shows that a simple and easy to operate single mobile phase TLC method can perform at the same level as the gold standard HPLC method for real clinical samples.