A High Performance Liquid Chromatographic method was developed and validated for quantitative determination of Guaifenesin impurities including 2-(2- methoxyphenoxy)propane-1,3-diol (β-isomer) and 2- m ethoxyphenol (guaiacol) in different multi drug components pharmaceutical dosage forms, containing guaifenesin, ambroxol hydrochloride and salbutamol sulfate . The different analytical performance parameters such as linearity, precision, accuracy, limit of detection (LOD), limit of Quantification (LOQ) were determined according to International Conference on Harmonization (ICH) Q2B guidelines. The chromatographic separation was achieved on EC NUCLEODUR-100-3C18 (250x4,6 mm, 5μm packing) column using gradient elution of Solvent A (0.1 M ammonium acetate buffer of pH 6.8) and solvent B (acetonitrile : methanol (80:20)) The Ultra Violet spectrophotometric determination was performed at 275 nm. The Linearity of the calibration curves for the analytes in the desired concentration range is good (r2 = 0.999) by High Performance Liquid Chromatography. The LOQ were 1 and 0.1 μg/ml respectively for guaifenesin β-isomer and guaiacol. The average percentage recovery of guaifenesin impurities was found to be within 98.6 – 101.2% of range. The developed method can be successfully used for identification and quantification of guaifenesin impurities β-isomer and guaiacol in the presence of guaifenesin, ambroxol hydrochloride and salbutamol sulfate in multi drug components pharmaceutical formulations.