Post on 13-Apr-2017
transcript
Botanical Description,Phytochemical
analysis&Analytical Profiles of palnt Carissa carandas Linn
Submitted by:D.GnanabhaskarRegno:16TMM2364
Under guidance of:Dr.Sanjay JachakDr.Inderpal Singh
Department of Natural ProductsNational Institue of Pharmaceutical Education and research
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Flow of TopicsAnalytical profiles
-HPLC profile-NMR profile
-Mass spectroscopy
Refernces
Conclusion
Introducton-Botanical profile-Therapeutic uses
-Chemical constituents
Isolation From roots,leaves,fruits,bark
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Introduction
A large evergreen shrub bark light grey. Branches have strong thorns
. Leave round and shiny About one inch in length.
Fruits are drupe type, ellipsoid, purplish black when ripe;arranged as a clusters.
seeds oblongoid, concave endosperm fleshy. Flowering and fruiting periods: October - January
Flowers small, scented. Similar in shape to jasmine. ebracteolate, pedicellate, bisexual, hypogynous
Current research found to be showing new therapeutic activites. Its leaves,fruits,had been used as Home remedies.
Name of the plant: Carissa carandas Linn. (Apocynaceae)
Parts of plants used medicinally: Root, Leaf, Fruit, Bark
Others names:Sanskrit : Karamla, KaramardakaHindi : Karaonda, KaraondiTelugu : Vaka, KaravandeMarathi : KarabandaEnglish : Karunda,Canberry
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Botanical profile
Kingdom Plantae Division Magnoliophyte Class Magnoliopsida Order Gentianales Family ApocynaceaeGenus Carissa Species C.carandas
Kinghorn, A. D. (2004). Quality standards of Indian medicinal plants, Vol. 1 A. K. Gupta
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Therapeutic usesAnti-inflammatory activity (N.Anupama, 2014) (Galipalli, 15 Dec
2014.)
Antioxidant Activity (Mahmud, 2010)
Anti diabetic activity (Prakash R. Itankar , 2011), ∗
Anticancer activity (Begum, 2013),
Anti diarrheal property, Laxative Effect (HassanMehmood, 2014).
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Phyto chemical constituents
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Extraction&isolation
Anti-inflammatory activity(J774
A.1 cells)
Roots coarsely powdered(3.47kg)
sequential extraction
Hexane(77.6gm) Ethyl acetate(18.78g) Methanol extracts(128.9gm)
Soxhlet apparatus for 24 h each (3 x 6 L)
fractionation using Vaccum liquid chromatography.
20-30 mesh
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After extraction filtered using
whatman filter paper then dried under
vacuum conditions
Cell culture, cell viability and nitric oxide release assay
Assays for measurement of TNF-α and IL-1β
Carissone(IC50=20.1 ± 2.69 μg/mL) Scopoletin (IC50=24.6 ± 1.36 μg/mL)
30 μM, were found to inhibit 41.88-53.44 % of TNF-α and IL-1β.
Galipalli, S., Patel, N. K., Prasanna, K., & Bhutani, K. K. (2014). Activity-guided investigation ofCarissa carandas(L.) roots for anti-inflammatory constituents. Natural Product Research, 29(17), 1670–1672. doi:10.1080/14786419.2014.989846
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Fractionation(VLC) (G2, 10.8 x 12.7cm) on silica gel (257 g, 230-400 mesh) with gradient elution using a solvent system; 0-50% EtOAc/Hex, 50-100% EtOAc/Hex and 0-20% MeOH/EtOAc, 20-50% MeOH/EtOAc, 50-70% MeOH/EtOAc and 70-100% MeOH
B1 (0.81 g) B2 (11.16 g) B3 (55.86 g) B4 (41.14 g) B5 (7.72 g)
Carissone(240mg)
Scopoletin(53mg)
Hex/EtOAc.Gradient elution
Based on invitro anti inflammatory activity&TLC profile
8Subfractions were formedB1.2-Stigmasterol(30mg)B1.3-LupeonlB1.7-Oleanolic acid
stepwise increase in polarity from 9:1::Hex:EtOAc to 7:3::EtOAc: MeOH.
12Subfractions were formedB2.5-Carissone(240mg)B2.9-Scopoletin(53mg)
Galipalli, S., Patel, N. K., Prasanna, K., & Bhutani, K. K. (2014). Activity-guided investigation ofCarissa carandas(L.) roots for anti-inflammatory constituents. Natural Product Research, 29(17), 1670–1672. doi:10.1080/14786419.2014.989846
Marker compound : Carissone
Sample preparation : Prepared in HPLC graded MeOH (5mg/ml) and filtered through 0.45µm Working solutuions are made by diluting with Stock solution with HPLC grade methnol (0.1,0.001,0.001mg/ml w/v) Chromatographic system : HPLC600Column details : Sun fire C18 Coloumn (4.6×250mm,5µm) Mobile phase : a)Acetonitrile, b)(0.4%)formic acid in water
Elution mode :Gradient elution was carried out starting from 80 To78% of B in4min,from 78% to75% Of B in 2 Min,from 75% to 72% of A in 1min, from 72% To 70% of B in 2 min,from 70 to 68% of B in 2min, from 68 to65% of B in 2min,and finally 65 to 50% of B in 2 min. Detection wave length: 265nm
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HPLC-Analysis
Galipalli, S., Patel, N. K., Prasanna, K., & Bhutani, K. K. (2014). Activity-guided investigation ofCarissa carandas(L.) roots for anti-inflammatory constituents. Natural Product Research, 29(17), 1670–1672. doi:10.1080/14786419.2014.989846
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Retention times
21.7-Carissone,
27.5- oleonolic acid
10.7- Stigmasterol,
11.2-Scopoleetin.
Detection wave length : 265nm
Galipalli, S., Patel, N. K., Prasanna, K., & Bhutani, K. K. (2014). Activity-guided investigation ofCarissa carandas(L.) roots for anti-inflammatory constituents. Natural Product Research, 29(17), 1670–1672. doi:10.1080/14786419.2014.989846
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1H NMR :
In the 1H NMR spectrum, 24 proton signals were
observed which supports the sesquiterpene type compound. The
characteristic singlet each for 3 protons at δ 1.20, 1.24, 1.26 and
doublets at 1.76 (J = 1.2 Hz) corresponds to four methyl signals
of eudesmane type skeleton.
Galipalli, S., Patel, N. K., Prasanna, K., & Bhutani, K. K. (2014). Activity-guided investigation ofCarissa carandas(L.) roots for anti-inflammatory constituents. Natural Product Research, 29(17), 1670–1672. doi:10.1080/14786419.2014.989846
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13C-NMRstudy of carissone
In the 13C NMR spectrum, 15 carbon
signals were observed; a carbon signal
at δ 199.31 corresponds to a keto
group. 4 Methyl signals were
observed at δ 27.46 & 26.59
(isopropyl unit), δ 22.43 and δ 10.89;
signal at δ 72.42 was assigned to
carbon containing hydroxyl group.
Rest of the carbon signals supported
the features of carissone.
Galipalli, S., Patel, N. K., Prasanna, K., & Bhutani, K. K. (2014). Activity-guided investigation ofCarissa carandas(L.) roots for anti-inflammatory constituents. Natural Product Research, 29(17), 1670–1672. doi:10.1080/14786419.2014.989846
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CONCLUSION
The fallowing researches are made on plant Carissa carandas are found to be showing different
potential therapeutic benfits. Ethnopharmacological studies strengthen the concept for utilizing C.
carandas plant as a source to facilitate safe and effective herbal treatments for biological problems
. Furthermore the aim is to provide a direction for further clinical research.
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1.Kinghorn, A. D. (2004). Quality standards of Indian medicinal plants, Vol. 1 A. K. Gupta, coordinator (medicinal plants unit,
Indian council of medical research). Indian council of medicinal research, Ansari Nagar, New Delhi, India.Journal of Natural
Products, 67(4), 739–740. doi:10.1021/np030714y
2. Galipalli, S., Patel, N. K., Prasanna, K., & Bhutani, K. K. (2014). Activity-guided investigation ofCarissa carandas(L.) roots
for anti-inflammatory constituents. Natural Product Research, 29(17), 1670–1672. doi:10.1080/14786419.2014.989846
3. Siddiqui, B. S., Ghani, U., Ali, S. T., Usmani, S. B., & Begum, S. (2003). Triterpenoidal constituents of the leaves of Carissa
Carandas. Natural Product Research, 17(3), 153–158. doi:10.1080/1478641031000104109
4. Mehmood, M. H., Anila, N., Begum, S., Syed, S. A., Siddiqui, B. S., & Gilani, A.-H. (2014). Pharmacological basis for the
medicinal use of Carissa carandas in constipation and diarrhea. Journal of Ethnopharmacology, 153(2), 359–367.
doi:10.1016/j.jep.2014.02.024
5. Anupama, N., Madhumitha, G., & Rajesh, K. S. (2014). Role of dried fruits ofCarissa carandasas anti-inflammatory agents
and the analysis of Phytochemical constituents by GC-MS. BioMed Research International, 2014, 1–6.
doi:10.1155/2014/512369.
REFERENCES
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6. Itankar, P. R., Lokhande, S. J., Verma, P. R., Arora, S. K., Sahu, R. A., & Patil, A. T. (2011). Antidiabetic potential of unripe Carissa carandas Linn. Fruit extract. Journal of Ethnopharmacology, 135(2), 430–433.
doi:10.1016/j.jep.2011.03.036 7.Mehmood, M. H., Anila, N., Begum, S., Syed, S. A., Siddiqui, B. S., & Gilani, A.-H. (2014). Pharmacological basis for the medicinal use of Carissa carandas in constipation and diarrhea. Journal of Ethnopharmacology, 153(2), 359–367. doi:10.1016/j.jep.2014.02.024.
8.Myanmar Medicinal Plant Database Family: Apocynaceae compiled by U Kyaw Tun, U Pe Than, and staff of TIL, http://www.tuninst.net/MMPD/TIL/famA/Apocynaceae.htm#Carissa-carandas
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For our research We are using so much plant material,,,..If our work fails then we are the cause of this much invasion
If our work Sucseeds then we are the cause for future conservation….
So we have to be thorough before stepping .,my friends