A simple and sensitive ultraviolet spectrophotometric method for quantitative estimation of glibenclamide in presence of lipid turbidity has been developed, to avoid false estimation due to diffraction by turbidity. UV detection has been performed at 232 nm, 227 nm and 237 nm and the calibration curve is plotted between resultant of absorbance of [232 nm - (227 nm + 237 nm)/2] and concentration of analyte. The calibration curve is found to be linear over the concentration range tested (2-20 mu g/mL) with Limit of Detection of 0.27 mu g/mL and Limits of Quantification 0.82 mu g/mL. Percent relative standard deviations and percent relative mean error, representing precision and accuracy respectively, for clear as well as turbid solutions are found to be within acceptable limits i.e. always less than 0.69 and 0.41 respectively for clear solution and 0.65 and 0.47 respectively for turbid solution. The recovery studies are also found to be within acceptable limits (100.10 to 101.21%) for both types of solutions. Conclusively, our method is successfully applied for the determination of glibenclamide in clear as well as turbid solution and it is found that the drug analyte in both types of solutions can be detected from the same calibration curve accurately and precisely and glibenclamide entrapped in the liposomes or in proliposomal matrix is not detected.