This document summarizes analytical profiles of the plant Carissa carandas Linn, including its botanical description, phytochemical analysis, and isolation of compounds. Key points include: - Carissone and scopoletin were isolated from root extracts and found to have anti-inflammatory activity. - HPLC, NMR, and mass spectroscopy were used to analyze marker compounds. HPLC showed retention times for carissone, oleonolic acid, stigmasterol, and scopoleetin. - NMR analysis supported the identification of carissone. 13C NMR showed signals for a keto group and hydroxyl group in carissone. - Several therapeutic activities of C

1. Botanical
Description,Phytochemical
analysis&Analytical Profilesof palnt
CarissacarandasLinn
Submitted by:
D.Gnanabhaskar
Regno:16TMM2364
Under guidance of:
Dr.Sanjay Jachak
Dr.Inderpal Singh
Department of Natural Products
National Institue of Pharmaceutical Education and research

2. Flowof Topics
Analytical profiles
-HPLC profile
-NMR profile
-Mass spectroscopy
Refernces
Conclusion
Introducton
-Botanical
profile
-Therapeutic
uses
-Chemical
constituents
Isolation
From
roots,leaves,
fruits,bark
2

3. 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, Karamardaka
Hindi : Karaonda, Karaondi
Telugu : Vaka, Karavande
Marathi : Karabanda
English : Karunda,Canberry
.
Botanical profile
Kingdom Plantae
Division Magnoliophyte
Class Magnoliopsida
Order Gentianales
Family Apocynaceae
Genus Carissa
Species C.carandas
3
Kinghorn, A. D. (2004). Quality standards of Indian medicinal plants, Vol. 1 A. K. Gupta

4. Anti-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).
4

5. Phyto chemical constituents
5

6. Extraction&isolation
6

7. 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
7
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

8. 8
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 formed
B1.2-Stigmasterol(30mg)
B1.3-Lupeonl
B1.7-Oleanolic acid
stepwise increase in
polarity from
9:1::Hex:EtOAc to
7:3::EtOAc: MeOH.
12Subfractions were formed
B2.5-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

9. 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 : HPLC600
Column 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
9
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

10. Retention times
21.7-Carissone,
27.5- oleonolic acid
10.7- Stigmasterol,
11.2-Scopoleetin.
Detection wave length
: 265nm
10Galipalli, 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

11. 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.
11
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

12. 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.
12Galipalli, 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

13. 13
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.

14. 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. 14
REFERENCES

15. 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
15

16. 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 tobe thoroughbefore
stepping .,my friends
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