Pakistan is enriched with natural flora comprising of unique wild and cultivated species utilized predominately by humans and animals. Bahawalpur region is in one of the nine divisions of the Punjab province in Pakistan established near the Sutlej River and comprises of many valuable species including grasses. Grasses possess remarkable economic and medicinal importance. All the cereal crops (maize, wheat, rice, barley) are also cultivated grasses. Many grasses are also used as a source of medicines as these are cheap, non-toxic and easily accessible. Hence, it is of keen interest to explore the native grasses of Bahawalpur region so that they can be used as a source of medicine and dietary fibre.In the present study, seventeen grasses were selected and their extracts were prepared in four solvents depending upon the polarity. Biological potential, elements and volatile compounds present in these extracts were evaluated. Initially, phytochemical tests were done to confirm the presence of different secondary metabolites. Total phenolic and flavonoid contents were determined spectrophotometrically which were detected in all extracts in varying concentrations. Highest phenolic contents were displayed in the methanol extracts of Ochthocloa compressa and Saccharum spontaneum (263.345 ± 16.35 mg GAE/g and 242.071 ± 20.63 mg GAE/g) while highest flavonoids contents were found in the ethanol extract of D. annulatum (47.023 mg QE/g) and the methanol extract of C. citratus (38.887 ± 2.19 mg QE/g). Antioxidant potential was determined via six different methods and the activity index was calculated. Results revealed that species which showed highest phenolic and flavonoid contents also exhibit highest antioxidant potential.Similarly, antimicrobial potential of all selected extracts was checked using disc diffusion assay which revealed somewhat weak to moderate antimicrobial potential against selected pathogens. So, cytotoxic potential was carried out to observe the toxicity potential of selected species. Among all species, O. compressa n-hexane extract showed highest cytotoxic potential with LC50 value of 2.80 µg/mL. All other species also displayed remarkable cytotoxic potential against brine shrimps. DNA damage protection assay was performed which revealed that ethanol and methanol extracts were more ix effective against DNA damage protection while acetone and n-hexane extracts did not show good activity. All inclusive, methanol extracts displayed significant biological potential as compared to the other extracts so they were further selected to examine antihemolytic potential of the plants. Methanol extracts of C. citratus (96.163 % inhibition), S. spontaneum (94.667 % inhibition), O. compressa (65.821 % inhibition) and D. annulatum (60.423 % inhibition) showed remarkable anti-hemolytic potential. Antihemolyticpotential of these species was found to be linked with the antioxidant potential and phenolic and flavonoid compounds. Strong anti-hemolytic and antioxidant activity of these plants suggests promising role in treating different diseases.Moreover, elemental profiling was done using ICP-MS and metabolomic studies were carried out using GC-MS. Results revealed the presence of seven essential and eighteen non-essential elements. Essential elements were found in decreasing order of potassium > magnesium > sodium > calcium > iron > manganese > zinc. Most of the non-essential elements were present within the permissible limit set by WHO which showed that the species are safe to use. GC-MS studies revealed the presence of more than 100 compounds in different concentrations. Decane (4.136 minutes) and m-cresol (5.46 minutes) were noted at minimum retention time in most of the species while Z-9hexadecenal (29.44 minutes) and 8, 11, 14-docosatrienoic acid, methyl ester (29.421 minutes) were identified at maximum retention time in most of the species. Total seventeen fatty acids and almost eighty-one different hydrocarbons were identified. Besides these, some monoterpenoids and diterpenoids were also detected in some species. Some of these compounds are known to possess cytotoxic potential while some of these compounds exhibit antimicrobial and antioxidant potential. Hence, these compounds are considerably involved in biological activities.In the end, individual organs of three species (S. halepense, D. annulatum and I. cylindrica) were investigated for further cell wall analysis. Acid-digestion of cell wall fractions revealed that xylose, arabinose, glucose and some traces of galactans are present more strongly in the aerial parts of selected species than the underground parts. Lichenase digestion confirmed that cellotrisoses exceeds cellotetroses and cellobioses in the cell wall of grasses. Dot-blot assay was done using eight commercial antibodies and comparison was made among dialysed and non-dialysed samples. Results revealed that x MLG, xylans, xyloglucans, arabinoxylan and glucoronoxylan were abundantly present in the sodium hydroxide extracts of non-dialysed samples while pachyman and mannan were not detected in significant concentrations. Some galactans were noted in the imidazole extracts of all fractions which showed the presence of pectins. Dialysed samples were not found effective to isolate hemi-celluloses from the fractions. Altogether, aerial parts possess more hemi-celluloses than the underground parts of plants.It is concluded that among all selected grass species, C. citratus, S. spontaneum, O. compressa and D. annulatum possess significant antioxidant, anti-hemolytic and cytotoxic potential. Overall, all species exhibited a variety of elements and compounds which are responsible for their biological activities. Moreover, these species can also be used as a biological marker. Cell wall analysis confirmed the presence of glycans in the aerial parts of plants which can be used as a dietary fibre and in different fibre industries. In future, in vivo studies should be carried out to confirm the medicinal value of these plants and compound isolation and microarray based profiling should be done to confirm the presence of non-volatile compounds and cell wall glycans.