PHYTOCHEMICAL AND PHARMACOLOGICAL INVESTIGATION OF PLANTS USED IN UNANI MEDICINE

Abstract

Unani system of medication is one of the oldest systems that prevails until date with its efficient drugs derived from plant, animal, and natural resources. In the Unani system of medicine, the drugs obtained from medicinal plants have a great interest due to their diverse applications. These plants comprise of various types of primary and secondary metabolites. Some of them have been playing a major role in the discovery of the new drugs for the treatment of various types of human and livestock diseases. Sisymbrium irio L is a medicinal plant and it belongs to the Family of Cruciferae and the various parts of this plant have been utilized as medicine and it is a world-wide distributed plant. Like Sisymbrium irio L, Suranjan Shireen (Colchicum autumnale L) which belongs to the family of the Liliaceae is primarily used for the treatment of arthritis in Unani medicine. The current investigation was carried out to analyze phytochemicals and pharmacological activities of Sisymbrium irio L and Colchicum autumnale L. The plant sample of Sisymbrium irio L and Colchicum autumnale L were bought from Khari Baoli Market, Delhi, Indian Drugs House, identified voucher ID 6238 & SC 0171/15 & extracted with n-hexane, dichloromethane, & methanol for 5-8 hrs in the Soxhlet apparatus. Elemental analysis was done by using ICP-MS. Isolation, Identification, and Investigation of compounds were determined by a hyphenated spectroscopic and chromatographic techniques such as, GC-MS, FT-IR, NMR, MS, UV-Vis spectrophotometer and UHPLC-Q Exactive Orbitrap. The pharmacological properties for instance, the antibacterial, antioxidant and anti dengue activity of Sisymbrium irio L and Colchicum autumnale L extracts were evaluated in vitro using paper disc diffusion, DPPH, ABTS, and MTT assay respectively. The qualitative phytochemical investigation demonstrated the presence of phenols, flavonoids, and terpenoids in every solvent extract of Sisymbrium irio L and Colchicum autumnale L, while steroids were recognized uniquely in n-hexane, dichloromethane and methanol extracts of the two plant extracts. However, saponins and tannins were not distinguished in any solvent extracts of these plants. Quantitative investigation of Sisymbrium irio L extracts affirmed that the methanol extract contained the most notable number of phenolic substances and flavonoid substance, subsequent dichloromethane, and n-hexane, while the dichloromethane extract of Colchicum autumnale L contained the highest number of flavonoid and phenolic substances, following methanol and n-hexane extracts. The phytochemical constituents are responsible for its antioxidant, antimicrobial and anti dengue activity and other medicinal uses. Antioxidant analysis examination confirmed that the methanol extract of Sisymbrium irio L demonstrated the most notable antioxidant scavenging activity in comparison with n-hexane and dichloromethane extracts, while dichloromethane extract of Colchicum autumnale L exhibited the uppermost percentage of free radicals scavenging activity in comparison to n-hexane and methanol extracts. Antibacterial activity demonstrated dose dependent activity on the tested bacterial strains. In vitro antiviral activity was performed against dengue virus-2 by the 3 (4, 5-dimethylthiazol-2-yl) 2, 5-diphenyltetrazolium bromide (MTT) strategy. Level of cell survival was likewise assessed by MTT. At this point of time maximum nontoxic dose (MNTD) of Sisymbrium irio L plant was examined through testing the dichloromethane, ethanolic, methanolic, and water extracts against Vero cells in vitro. Antiviral test dependent on cytopathic effects indicated via the level of inhibition upon treating DENV infected Vero cells with a maximum nontoxic dose (MNTD) of Sisymbrium irio L has the most antiviral inhibitory effects. ICP-MS elemental analysis of Sisymbrium irio L and Colchicum autumnale L displayed the existence of Na, Mg, K, Ca, Sr, Mn, Zn, Al, P and Fe. GC-MS, and UHPLC-Q Exactive Orbitrap investigation of the extracts confirmed existence of:- Hexadecanoic acid, methyl ester, n-Hexadecanoic acid, (E) - 9 - Octadecenoic acid, ethyl ester, 9, 12 - Octadecadienoic acid (Z, Z), Linoleic acid ethyl ester, β sitosterol, Isorhamnetin, Isorhamnetin-3-neohesperidine, Isorhamnetin - 7 - O - beta - D - glucopyra noside, Isorhamnetin - 7 - glucoside, Isorhamnetin-3-Laminaribioside, Colchicine, (R/S)-Deacetyl Colchicine, 3-demethyl Colchicine, and Colchicoside (3-demethyl colchicine glucoside). The anti inflammatory activity suggests that Colchicum autumnale L can be used for relieving the symptoms of inflammation. Comparative docking studies revealed colchicoside as the most potent anti inflammatory compound with minimum binding energy (affinity) as compared to other compounds and standard drug diclofenac. Bioassay-guided effort yielded a compound which after characterization using UHPLC-QExactive Orbitrap, MS, 1H-NMR, 13C-NMR, and FT-IR identified as a class of apigenin, isorhamnetin derivative, and colchicine derivative. These spectroscopic techniques play an indispensable function in the isolation, identification & investigation of natural products for the discovery of new drugs. Hence, phytochemical analysis through the above spectroscopic techniques confirmed that Sisymbrium irio L and Colchicum autumnale L extracts contained the various types of bioactive compounds, and these bioactive compounds have an important role in the therapeutic system of medicine for treatment of disease. Further, Sisymbrium irio L and Colchicum autumnale L must be explored more on in vivo test in order to determine the mechanisms of action.