A novel UV-Spectroscopy method was developed and validated for the estimation of L-glutathione in bulk and dosage form with congo red. The wavelength at which L-glutathione and congo red mixture product showed maximum absorption at 568nm in distilled water. The developed method range was found to be10-90µg/ml. The developed method validated for linearity, precision, range, limit of detection, the limit of quantification, robustness, specificity, system suitability. The regression equation was found to be y = 0.0045x + 0.121 with correlation coefficient R² = 0.9983. The limit of detection and the quantification limit was 5.05 µg/ml and 15.32 µg/ml respectively. The developed method was found to be linear, precise, robust, economical for the evaluation of L-glutathione in bulk and dosage form.