Sundhararajan, R (2012) Phytochemical and Pharmacological Screening of Indigenous Medicinal Plants. Doctoral thesis, The Tamilnadu Dr. M.G.R. Medical University, Chennai.
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Abstract
In the present investigation, pharmacognostical, phytochemical and pharmacological studies was taken up on the whole plant of Spermacoce hispida and Spermacoce ocymoides. The significant observations made during the study include: The present investigation documented the pharmacognostical study of S.hispida and S. ocymoides, which comprises of macroscopical, microscopical, powder microscopy and determination of physico-chemical constants like ash values, extractive values, loss on drying and estimation of inorganic constituents. These parameters serves as standards providing an imperative basis of information in ascertaining the uniqueness and genuinity to decide on the quality and clarity of the plant material, further serve as an identity tool in differentiating S.hispida and S.ocymoides and other species or varieties of the genus Spermacoce. From the present investigation it is evedent that the phytochemical study of n-Hexane, ethyl acetate and ethanolic extracts of S.hispida and S.ocymoides were analyzed for the presence of alkaloids, carbohydrates, glycosides and anthraquinones, flavanoids, tannins and phenolic compounds, proteins and amino acids, saponins, sterols and or triterpenes. In this study SHE and SOE had showed the presence of numerous phyto-constituents when compared to the other extracts. HPTLC finger print of n-hexane, ethyl acetate and ethanolic extracts S.hispida and S.ocymoides were analyzed, in this SHH, SHEA and SHE had showed 10, 8 and 11 peaks respectively with maximum peak area of 40.44% in SHE. Similarly, SOH, SOEA and SOE had showed 12, 12 and 10 peaks respectively with maximum peak area of 38.36 in SOH and 34.10 in SOE. GC-MS analysis of ethanolic extract of S.hispida had revealed 57 components, in which 9 components showed maximum peak area. GC-MS analysis of ethanolic extract of S.ocymoides had revealed 62 components, in which 8 components showed maximum peak area. In the preliminary anti-inflammatory activity ethanolic extract of S.hispida and S.ocymoides produced maximum response. Hence, due to the potential response evidenced with that of the ethanolic extract, the isolation of compounds were performed on SHE and SOE using column chromatography. In SHE, two components were isolated; kaempferol and erythrodiol and in SOE one compound was isolated; ursolic acid. The compounds were identified based on the spectral studies (IR, 1H NMR, 13C NMR and Mass Spectra) and the structure was elucidated. The pharmacological studies on the plant S.hispida and S. ocymoides, which includes acute toxicity study, anti-inflammatory, analgesic, antipyretic, invitro and in-vivo antioxidant activity. In the acute toxicity n-Hexane, ethyl acetate and ethanolic extracts of S.hispida and S.ocymoides were declared and found to be safe upto the dose of 2000 mg/kg p.o. The preliminary anti-inflammatory activity of n-Hexane, ethyl acetate and ethanolic (95%) extracts of S.hispida and S.ocymoides was evaluated using carrageenan induced rat paw edema. In the present study both the ethanolic extract of S.hispida and S.ocymoides displayed significant inhibition of paw volume when compared to other extracts studied. Isolation of active biological component was carried out with the ethanolic extract due to its potential activity, resulted in the isolation of kaempferol and erythrodiol from S.hispida and ursolic acid in S.ocymoides. Hence, further pharmacological studies was proceeded with SHE, kaempferol, erythrodiol, SOE and ursolic acid. In carrageenan induced rat paw edema and cotton pellet induced granuloma erythrodiol (80 mg/kg) and ursolic acid (80 mg/kg) evidenced significant anti-inflammatory activity when compared to the other compounds. The analgesic activity was screened by acetic acid induced writhing reflex and tail immersion model. In acetic acid-induced abdominal constrictions SHE and SOE showed maximum effect, whereas in the tail immersion model erythrodiol and ursolic acid exhibited maximum protection. SHE, kaempferol, erythrodiol, SOE and ursolic acid had not alters the normal body temperature, whereas SHE and SOE documented significant anti-pyrexia activity in yeast induced pyrexia model when compared to the isolated compounds. COX inhibitory assay: It is very much incidentally correlative from the present investigations on the plant that the potential anti – inflammatory activity exhibited by the extract as well as the isolated bioactive compounds could be mediated via inhibition of COX - 1 and 2 enzymes either partially or on the whole. These findings substantiate the traditional claim documented on the plant for treating the inflammatory condition. In the present investigation on the plant S. hispida and S. ocymoides, where kaempferol and ursolic acid showed marked COX 2 activity and with respect to COX 1 activity there was a considerable response observed with kaempferol alone. Hence it may be said that kaempferol exhibited both COX 1 and 2 more effectively. On the contrary, with COX 1 activity being negligible with sample erythridiol and ursolic acid may be justified to show a pronounced efficient activity restricted to COX 2 inhibition. The kaempferol isolated showed both COX 1 and 2. Ursolic acid showed more selectivity. The erythridiol isolated from the plant on COX inhibition assay was negligible. The vitro antioxidant activity of SHE, kaempferol, erythrodiol, SOE and ursolic acid was evaluated using various models: In DPPH free radical scavenging assay kaempferol and ursolic acid showed significant free radical scavenging activity, whereas in nitric oxide radical inhibition assay SHE and SOE exhibited significant nitric oxide radical quenching activity, similarly in hydroxyl radical scavenging assay model kaempferol and ursolic acid with remarkable activity. In the invivo antioxidant activity, SHE and SOE elicited maximum activity was evident with increase in the level of antioxidant enzymes viz SOD, CAT and Gpx and decrease in the level of MDA.In this study effect of extracts may be due to the presence of multiple components in the extract apart from kaempferol, erythrodiol and ursolic acid. It is evident from the present study that the plants S.hispida and S. ocymoides expressed potent anti-inflammatory, analgesic, invitro and in vivo anti-oxidant potentials. In addition the plants expressed remarkable Cyclooxygenase – Inhibition Assay which substantiate the results obtained in the inflammatory models studied. Interestingly, the pharmacological responses produced in the invitro Cyclooxygenase – Inhibition study recommends the emphasize potential of the plant S. hispida as the results are more significant in both Cyclooxygenase –1 and 2. CONCLUSION: It is concluded from the present study on the plants S.hispida and S.ocymoides belonging to the family Rubiaceae revealed its identity as a potential source in treating inflammatory conditions and the same was further significantly confirmed by Cyclooxygenase – Inhibition Assay studied on both the ethanol extracts of the plants and their isolates namely, Kaempferol, Erythrodiol and Ursolic acid. Further more, the ethanol extract and their isolates demonstrated equally potential analgesic, anti-pyretic and invivo/invitro anti-oxidant potentials of the plants. The bioactive isolate Kaempferol was first reported to be identified in the plant S.hispida. Similarly, Ursolic acid was reported for the first time in the plant S.ocymoides. Future focus on extended research with respect to the bioactive isolates may pave a way to identify and explore the phytochemical potential of the plants with respect to the management and treatment of inflammatory conditions along with its analgesic, anti-pyretic and anti-oxidant activities. The pharmacognostical characters documented on the plants may serve as a typical standard in ensuring the plants future authenticity.oxicity
| Item Type: | Thesis (Doctoral) |
|---|---|
| Uncontrolled Keywords: | Phytochemical, pharmacological screening, indigenous medicinal plants. |
| Subjects: | PHARMACY > Pharmaceutics |
| Depositing User: | Subramani R |
| Date Deposited: | 20 Aug 2017 08:48 |
| Last Modified: | 28 Oct 2022 14:49 |
| URI: | http://repository-tnmgrmu.ac.in/id/eprint/2736 |
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