carissa carandas plant botanical profile, analytical profile, isolation profiles of plant active constituents

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)


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


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Retention times

21.7-Carissone,

27.5- oleonolic acid

10.7- Stigmasterol,

11.2-Scopoleetin.

Detection wave length : 265nm


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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.


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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.


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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