In villages farmers often use synthetic hormones and antibiotics without suggestion of medical practitioner. Government restricted the usage of synthetic hormones without prescription of medical practitioner but farmers use these hormones for enhanced production of milk in cows and buffaloes. As of now there is no scientific approach for the detection of usage of synthetic hormones in animals. HPLC, GC, LCMS, GCMS and ELISA are the widely using methods to detect or quantify hormones. Unfortunately these methods cannot give us a clue regarding the origin of hormone. As synthetic hormones have preservatives, the administrated hormones will be utilised by animals and the preservatives will be excreted by animal through dung and urine.Preservatives are the chemicals that are used in drug products to prevent decomposition by microbial growth or by undesirable chemical changes and detection of these preservatives may give us a clue on usage of synthetic hormones. Our GCMS analysis of HPLC fractions proved that these preservatives are excreting in dung and our analysis found Benzoic acid, Benzothiazole, Benzoyl-2-ethoxymethyl benzene, Benzyl Benzoate, Glycerol, Levulonic acid, Phenazinol, Phenol, Propanoic acid and Succinic acid in Dung extracts. Our study provided a clue to investigate synthetic hormones by assessing preservatives using dung and it can be further investigated to evaluate the level of synthetic hormone and medicine load on cattle and their impact on human health.