Phytochemical studies of medicinal plants from several genera with antidiabetic activities

Karla M. Rodrguez Tirado Mentor: Dr.Jannette Gavilln May 13th , 2011

Acknowledgements

Chemistry Department Technicians Juan Carlos Rodrguez Jannette Gavilln, PhD

Alkaloids

Tannins Cardiac Glycosides Sterols Phenols

Terpenoids

Flavonoids Saponins

Previous Phytochemical Studies

Recent research have demonstrated the ability some phytochemicals have in protecting humans and of being useful for the treatment of diseases.
Rupasinghe et al. (2003) have reported saponins antidiabetic properties ( Kumar et al. 2009) Studies have also shown that glycosides, flavonoids, tannins and alkaloids have hypoglycemic activities ( Kumar et al. 2009) Terpenoids have also been shown to decrease blood sugar level in animal studies (Kumar et al. 2009) Several plant derived flavonoids have been reported to inhibit aldose reductase activity and impart beneficial action in diabetic complications ( Tiwari and Rao 2002)

Our plants

S.jambos- Tannins have been reported to be a mayor compound.

Saponins also reported in lower amounts (Djadjo et al. 2000 ) Flavonoids and Phenolic compounds have also been reported (Reynertson et al, 2008)

C.spiralis- previous studies reveal the occurrence of sterols, glycosides and saponins (da Silva and Parente 2004) R.spathacea- has been reported to be rich in flavonoids, triterpenes and phenolics compounds (Rosales-Reyes et al., 2006 )

Specific Aims
Use thin layer chromatography (TLC) to identify the phytochemical profile of C. spiralis, T. ananassae, S. jambos and R. spathacea To complete the qualitative analysis of Alkaloids and Flavonoids for the plants under study To compare the qualitative and TLC results and draw conclusions

Relevance
At the moment, there is no phytochemical profile for these plants to which people attribute antidiabetic properties. If we knew the phytochemical profile, we would be able to identify those specific phytochemicals responsible for the antidiabetic properties. These plants could then be use as complementary or alternative medicines.

Methodology Qualitative Experiment

Flavonoids Shinoda Test
Dried methanol extracts of plants

Add 95% methanol, drops of concentrated HCl and 0.5 g of magnessium

A pink color will reveal a positive result

Methodology Qualitative Experiment

Alkaloids
Prepare plant extracts

To the filtrate, add drops of Wagners Reagent A redddish-brown precipitates indicates a positive result

Methodology- TLC
Extraction procedures according to phytochemicals
T LC

S. jambos T. anassae R. spathaceae C. spiralis

UV264 and 366 Visible Light Dragendorffs Reagent Folin-Ciocalteus Reagent Iodine Vapors Anasaldehydesulphuric acid

on i t iza l a u Vis

Solvents: CH2Cl2 and Methanol Acetic Acid, Methanol and Water CH2Cl2, glacial acetic acid, Water and Methanol

Qualitative Results

Tan Tannins Alk- Alkaloids Sap- Saponins Fla- Flavonoids

CG- Cardiac Glycosides Ter- Terpenoids Ste- Sterols

Results Flavonoids

Standard: Quercetine Extraction: Methanol Liquid- liquid Extraction with a mixture of water and ethyl acetate Mobile Phase: CH2Cl2 and methanol Visualization: UV254nm and visible light

The TLC confirms the qualitative test result.

Rhoeo spathacea is the only plant that contains flavonoids. The result is confirmed by the literature.

Results- Alkaloids

Standard: Nicotine Extraction: NH4OH, lixiviate with EtOAc, add NH4OH to organic phase, extract with CH2Cl2

Mobile phase: CH2Cl2 and methanol Visualization: Spray with Dragendorffs reagent, UV254nm and visible light

Both, S. jambos and T.ananassae, contains alkaloids. This results compare to those obtain in the qualitative test.

The lane for R.spathaceae reveals the presence of alkaloids, differing from the qualitative test result.

Results- Phenolics

Standard: Hydroquinone Extraction: Lixiviate with methanol and condense filtrate Mobile phase: CH2Cl2 and methanol Visualization: Spray with Folin-Ciocalteus reagent and heat plates

The presence phenolic compounds is observed in every plant extract.

Tannins are phenolic compounds, therefore, this results confirm those obtained for the qualitative test in which every plant had a positive result. Literature confirms the positive results for S.jambos and R.spathacea.

Results: Sterols

Standard: Stigmasterol Extraction: Methanol and condense filtrate Mobil phase: CH2Cl2, glacial acetic acid, methanol and water Visualization: Spray with a solution of FeCl3, acetic acid and sulfuric acid and heat the plate

Sterols are present in every plant extract.

The results differ from the qualitative test results. But, previous research have confirm the presence of sterols in C.spiralis.

Results- Cardiac Glycosides

Standard: Digitoxin Extraction: 70%EtOH on rotary shaker, centrifuged 2 times adding 70% lead acetate and 6.3% Na2CO3, respectively, redesolve with CH2Cl2 Mobile phase: EtOAc-MeOH-H2O Visualization: Sulphuric Acid Reagent and UV366nm

The phytochemical is observed to be present in all of the plant extracts.

Confirm the qualitative results for R.spathaceae, C. spiralis and T.ananassae. This is not so for S. jambos, which had a negative for the qualitative. The presence of this phytochemical in C. spiralis is recorded by previous research.

Alkaloids:

Conclusions

The presence of alkaloids was observed for all plants studied R. spathaceae and C.spiralis showed higher amount of spots than T.ananassae and S.jambos.

Phenolics:
The presence of phenolics was observed for all plants studied S.jambos presents the most amount of spots this phytochemical.

Sterols and Cardiac Glycosides:

The presence of sterols and cardiac glycosides was observed for all plants studied These phytochemicals seems to be more or less in the same proportion for all plants.

Future Work
Complete qualitative analysis for phenolics compounds, sterols , terpenoids, Costus spiralis and Syzygium jambos plant extracts. Complete TLC of saponins, tannins and terpenoids.

References
Wagner R and Bladt S, Plant Drug Analysis, A Thin Layer Chromatography Atlas, 2nd Ed: Springer;Berlin, 1996. Leach MJ. Gymnema sylvestre for Diabetes Mellitus: A Systematic Review. J Altern Complement Med. 2007 Nov; 13(9):977-83 Tiwari AK, Rao JM: Diabetes mellitus and multiple therapeutic approaches of phytochemicals: present status and future prospects. Curr Sci, 2002, 83, 3038. Mallikharjuna PB, Rajanna LN, Seetharam YN, Sharanabasappa GK. Phytochemical Studies of Strychnos potatoruna L.f.- A Medicinal Plant. Journal of Chemistry. 4 (4): 510-518, 2007. Djadjo C, Delme M, Quentin-Leclercq J, (2000). Antimicrobial activity of bark extracts of Syzygium jambos(L.) Alston(Myrtacea). Journal of Ethnopharmacology, Volume 71, Issues 1-2, July 2000, Pages 307-313 Reynertson, K. A., Yang, H., Jiang, B., Basile, M. J., & Kennelly, E. J. (2008). Quantitative analysis of antiradical phenolic constituents from fourteen edible Myrtaceae fruits. Food Chemistry, 109(4), 883-890. doi:10.1016/j.foodchem.2008.01.021 Rosales-Reyes, T., de la Garza, M., Arias-Castro, C., Rodrguez-Mendiola, M., Fattel-Fazenda, S., Arce-Popoca, E., HernndezGarca, S., Villa-Trevio, S.Aqueous crude extract of Rhoeo discolor, a Mexican medicinal plant, decreases the formation of liver preneoplastic foci in rats. Journal of Ethnopharmacology, Volume 115, Issue 3, 12 February 2008, Pages 381-386 da Silva, B., & Parente, J. (2004). New steroidal saponins from rhizomes of Costus spiralis. Zeitschrift Fr Naturforschung. C, Journal Of Biosciences, 59(1-2), 81-85. Retrieved from EBSCOh Kumar A, Ilavarasan R, Jayachandran T, Decaraman M, Aravindhan P , Padmanabhan N, Krishnan MRV. Phytochemicals Investigation on a Tropical Plant, Syzygium cumini from Kattuppalayam, Erode District, Tamil Nadu, South India. Pakistan Journal of Nutrition 8 (1): 83-85, 2009.

Thanks for the attention