||Diabetes mellitus (DM) is a chronic metabolic disorder arises from deficiency (T1DM) and/or efficiency (T2DM) of insulin. T2DM accounts for more than 90% of all diabetics and its prevalence is increasing alarmingly worldwide. It is intimately associated with improper utilization of insulin by target cells and tissues. Insulin, a polypeptide hormone synthesized by the pancreatic beta cells, is responsible for the maintenance of glucose homeostasis in human and other mammals. It is essential for the entry of glucose across the muscle and adipocyte cell membranes for energy production, hepatic glycogen synthesis, protein and nucleic acid synthesis and inhibition of gluconeogenesis, glycogenolysis and lipolysis. Thus, T2DM is a multifactorial, multisystemic endocrine disorder for which monotherapy often fails as the disease progress to later stages. Traditional medicinal plants used for the treatment of DM contain various biologically active ingredients which act in a synergistic way in maintaining normal glycemia. However, only few of them have been subjected to scientific validation. One such medicinal plant which lacks scientific scrutiny is Piper betle leaves. Hence, in the present study an attempt has been made to extract and identify the chemical nature of biologically active phytochemicals present in the distinct variety of Piper betle leaves of South India. The leaves cultivated in Kumbakonam are known for their taste, quality and medicinal properties. The data obtained by HPLC analysis and spectral studies such as FTIR, Mass, 1H NMR, 13C NMR evidenced that the ethanolic extract of Piper betle leaves contains Caffeic acid, p-Coumaric acid, Eugenol, Rutin and Hydroxychavicol as major secondary metabolites which are known for their beneficial and pharmacological properties. The results of the present study suggest that the betel leaves are the rich source of pharmacologically important lead molecules and also provide the scientific rationale for the use of Piper betle leaves in the traditional medicine.