PHYTOCHEMICAL ANALYSIS AND GC-MS PROFILING OF BIOACTIVE COMPOUNDS IN Apis mellifera BEE PROPOLIS FROM INDIA

Bee propolis is a complex resinous substance collected by bees from plants and used as a construction material and protective substance in the hive. Propolis is a complex mixture of different compound classes; among them are the volatile compounds responsible for conferring the aroma and biological activity. In the present study, the volatile fraction of the Apis mellifera bee propolis samples was determined using Gas chromatography-mass spectrometry (GCMS). Crude propolis samples were obtained from five different phytogeographical regions of India namely Rohtas from Bihar, Latur from Maharashtra, Sawai Madhopur from Rajasthan, Rewari from Haryana, and Kota from Rajasthan. The physical parameters of the propolis samples were examined and noted. The samples showed variations in their physical appearance, color, and odor. The extraction of crude propolis with 70% ethanol yielded the dried propolis ethanolic extracts which showed a sticky and semi-solid appearance. The percentage yield (w/w) of ethanolic extracts of propolis was calculated. The highest extraction yield was recorded in the propolis sample from Rohtas, Bihar (83.50%) whereas the least extractive value was found in the propolis sample from Rewari, Haryana (11.60%). Qualitative analysis was carried out to identify the different classes of secondary metabolites in various propolis extracts. The findings demonstrated significant indication about the presence of various compounds like alkaloids, carbohydrates, terpenoids, phenols, flavonoids, tannins, phlobatannins, cardiac glycosides, steroids, and saponins. Various secondary metabolites like phenols, flavonoids, tannins, and alkaloids were quantified using standard protocols. However, the amount of these metabolites varied in different samples based on the region of collection. The results of quantitative estimation showed that the propolis sample from Sawai Madhopur, Rajasthan reported the highest contents of total phenols (77.30µg/g), total tannins (260µg/g), and total alkaloids (624µg/g) as compared to other samples. The Propolis sample from Rewari, Haryana (8.18 µg/g) recorded the highest flavonoid content than other samples. GC-MS technique was used for the profiling of volatile components in the samples. The results led to the identification of 40 compounds in the propolis samples with various pharmacological activities. Among the various chemical compounds identified in the propolis extracts, two novel bioactive compounds namely deoxy- podocarpol and p-1-indanyl phenol were reported for the first time. The results showed that Indian propolis could prove to be a potent source for the exploitation of these phytochemicals beneficial in the pharmaceutical and alternative medicine industries. It was also predicted from the study that the volatile constituents identified in Indian propolis have the potential to be developed into pharmaceutical drugs.