Paper Title
CHARACTERIZATION AND BIOLOGICAL EVALUATION OF CHEMICAL CONSTITUENTS FROM PINUS GERARDIANA, INULA RACEMOSE, ACONITUM HETEROPHYLLUM AND PIERORHIZA KURROA EXTRACTS

Abstract
AIM: Characterization and Biological evaluation of chemical constituents from Pinus gerardiana, Inula racemose, Aconitum heterophyllum and Pierorhiza kurroa extracts. Methods: The evergreen plants Picrorhiza kurroa (Kadu), Pinus gerardiana (chilgoza), Inula racemosa (Puskarmoola), and Aconitum hterophyllum (Ativisa) with numerous health benefits including antiseptic, anti-fungal, anti-bacterial, antioxidant, anti-inflammatory, etc. properties. They have been chosen to study the chemical compositions and health benefits. The extractions were done by Soxhlet apparatus using solvent such as n-hexane, chloroform, ethylacetate, acetone, n-butanol, methanol, hydroalcoholic and water and total 8 extracts were prepared. The primary identification of the compounds was done using LCMS and the, secondary identification was achieved by HPLC and HPTLC for the quantification of the compounds present in the extracts and findings have been reported as well. In-vitro Antimicrobial and antioxidant activity was selected for biological activity. Results: The primary identification of the compounds was done using LCMS. From HPLC and HPTLC analysis, total of six natural compounds in P. kurroa, eight natural compounds in I. racemoa, eight natural compounds in Pinus gerardiana, and three natural compounds in A. hetrophyllum were identified and quantified. Furthermore, the antimicrobial activity assays the maximum biological activity of the four plants P. kurroa was best, and among its eight extracts acetone extract showed good potency in terms of zone of inhibition in milimetere (11.66 ±1.52) against S. aureus, n-butanol showed (12.01±1.00) against S. mutans, acetone extracts showed (11.66±1.52) against E. coli, acetone extracts showed (11.33±1.52) against K. pneumonia and acetone extracts showed (11.00±1.00) against P. aeruginosa. In I. racemoa, ethyl acetate extract showed good potency in terms of zone of inhibition in milimetere (13.33±1.52) against S. aureus, -hexane extract showed (12.00±2.00) against S. mutans, acetone extracts showed ( 16.66±.47) against E. coli, acetone extracts showed (15.66±10.47) against K. pneumonia and methanol extracts showed (15.66±0.47) against P. aeruginosa. In Pinus gerardiana, hydroalcholic extract showed good potency in terms of zone of inhibition in milimetere (14.33±10.52) against S. aureus, acetone extract (16.01±1.81) against S. mutans, ethyl acetate extracts showed (16.33‡.042) against E. coli, ethyl acetate extracts showed (17.35±0.58) against K. pneumoniae and -hexane extracts showed good potency (16.33‡0.58) against P. aeruginosa. In A. heterophyllum, the methanolic extract showed good potency in terms of zone of inhibition in milimetere (11.00±1.00) against S. aureus, petroleum ether extract showed (11.66±1.52) against S. mutans, n-butanol extracts showed (12.33‡1.52) against E. coli, chloroform extracts showed (11.66±1.52) against K. pneumonia and methanolic extract showed (11.66±1.52) against P. aeruginosa. Antioxidant activity was found to be average against ascorbic acid. Conclusion: Four endangered plant were chosen for the study including Picrorhiza kurroa, Inula racemosa, Pinus gerardiana and Aconitum heterophylum. Twenty-three natural compounds were identified and quantified from four plant extracts of those plants. Primary identification was done by LCMS and matched with different databases. Secondary identification was done by HPLC and HPTLC. Antimicrobial and antioxidant activity was examined for eight extracts of four plants. The hydrogen and hydrophobic interaction were analyse for suitable molecule with IHNJ receptor of E. coli bacteria using docking study. Keywords - IR, LCMS, HPLC, HPTLC, Chloramphenicol, Ascorbic Acid, Antimicrobial, Antioxidant