The present invention provides a novel process for preparation of crystalline atropine sulphate monohydrate by esterification reaction of tropine hydrochloride with acetyl protected acid chloride of DL- Tropic acid in organic solvent in inert atmosphere. The condensed acetyl atropine hydrochloride is hydrolyzed in dilute hydrochloric acid to get atropine hydrochloride in aq. Medium which is basified by liquor ammonia and extracted in methylene dichloride and purified in acetonitrile to get good yield and quality of pure atropine. Atropine is then subjected to quaternarization to give its quaternary salt like Atropine sulphate.