Herbal formulations are widely used to treat a variety of diseases with the use of natural ingredients and have the additional advantage of negligible side effects. Looking towards the wide applicability of herbal formulations, a novel herbal formulation containing Tulsi and Parijat has been formulated and evaluated for different standardisation parameters. Tulsi and Parijat both possess multiple natural effects like antibacterial, antiviral and antipyretic properties along with wide applicability to treat different respiratory problems. Therefore, the combination of these two naturally available herbs can provide good immunomodulatory action for humans. Apart from the various phytoconstituents present in these two herbs, eugenol and tannic acid are found to have versatile effects. They are widely used to treat various common health issues like tonsillitis and various respiratory complaints like cough, cold and sour throat. A successful attempt has been made to develop the HPTLC method for the simultaneous determination of Tannic acid and Eugenol in pure and newly formulated churna containing Tulsi and Parijat as chief herbs. Tannic acid and eugenol mixtures get resolved by toluene:ethyl acetate:formic acid (5:3:2) with Rf of 0.849 ± 0.003 and 0.534 ± 0.009 respectively for tannic acid and eugenol. Linearity is observed within the range of 250–1500 ng/band for both eugenol and tannic acid. According to ICH Q2 R1 advice for analytical method validation, this designed technique has been effectively validated.