A rapid, simple, precise, accurate and reproducible reverse phase high performance liquid chromatographic method was developed for the estimation of mesoindigo in the bulk by using thermoscientific column (5 µm, 250 mm × 4.6 mm i.d). Combination of methanol: buffer (0.1% ammonium dihydrogen phosphate adjusted to pH-3 using orthophosphoric acid) in the ratio of 90:10 v/v as a mobile phase at a flow rate of 1 ml/min in an isocratic mode at a detection wavelength of 265 nm was used for the analysis. The drug was eluted at a retention time of 7 min. The method was validated as a final verification of method development with respect to precision, linearity, accuracy and robustness. Calibration curve was linear with the correlation coefficient of 0.999 over a concentration range of 50-300 µg/ml. The mean recovery obtained was within the range of 98-102%.
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