Quantitative analysis ofphytochemicals in antidiabeticplant extractsNatalie Rivera OrtizMentor: Dr. Jannette Gavillán Suár...
AcknowledgementsThanks to: Dr. Jannette Gavillán Suárez, Ph.D.
What are phytochemicals? They are secondary metabolites that occurnaturally in plants. Molecules responsible for the col...
Classification of PhytochemicalsAlkaloidsNicotineCardiac glycosidesOleandrin
Classification of PhytochemicalsSaponinsSolanineTerpenoidsUrsolic AcidIsoprene
Classification of PhytochemicalsPhenolsFlavonoidsQuercetin
Previous Studies Qualitative experiments have demonstrated thepresence of flavonoids, terpenoids, saponins, tanninsand ca...
Previous Studies Alkaloids, glycosides, carbohydrates, and steroidshave demonstrated activity consistent with theirpossib...
Goal and ObjectivesObjectives:Quantify flavonoids, total phenolic compoundsand saponins in plant extracts from Tapeinochui...
Relevance of this Research Quantification of phytochemicals will be keyin identifying biomarkers of antidiabetic activity...
MethodologyFlavonoidsQuercetinsolution(0.27mg/mL)/plantextracts , 200μL of AlCl310%and 200 μLof CH3COONa1MQuercetinCalibra...
MethodologyTotal Phenolic ContentQuercetinsolution (3.38mg/mL )/plantextracts, 600 μLof Lowryreactive C and200 μL of Lowry...
Results - Flavonoidsy = 34.8x - 0.055R² = 0.9940.00000.10000.20000.30000.40000.50000.60000.70000.80000.00E+00 5.00E-03 1.0...
Results - FlavonoidsConcentration of plantextract in assay:C1V1=C2V2C2 = C1V1/V2C2 = (0.100mL)(0.04094 g/mL)(5.0000 mL)C2 ...
CalculationsResults - FlavonoidsFlavonoid content by dry weight:(0.0224 mg/mL QE)(1mL) = 29.8 mg QE/g DWFlavonoid content ...
Results - FlavonoidsPlant Concentrationof flavonoids(± SD mgQE/ g DW)T. spathacea 13.7 ± 0.7C. speciosus 15.8 ± 0.9S. jamb...
Results – Total Phenolsy = 4.63x + 0.16R² = 0.9900.00000.10000.20000.30000.40000.50000.60000.70000.80000.90001.00000.00 0....
Results – Total PhenolsConcentration ofmethanolic extract inassay:C1V1=C2V2C2 = C1V1/V2C2 = (0.150μL)(3.00 mg/mL)(1500 μL)...
Results – Total PhenolsFlavonoid content in methanolic extract:(0.085 mg/mL QE)(1mL) = 0.29 mg QE/mg extract0.300 mg extra...
Results – Total PhenolsAqueousextractTotalPhenolicContent(± SD mgQE)T. spathacea 0.26 ± 0.01C. speciosus 0.35 ± 0.01S. jam...
Results – Total Phenols0 5 10 15S. jambosT.anannassaeC. speciosusT. spathaceaTotal Phenolic Content (mg QE/mg extract)Plan...
Current WorkConcentratedsaponins areextracted twicewith equalvolumes ofn-butanolDNS MethodQuantitativedeterminationof gluc...
Current Worky = 7.390x - 0.003R² = 1.0000.00000.20000.40000.60000.80001.00001.20001.40000.000 0.020 0.040 0.060 0.080 0.10...
Current WorkSugar content inQuijalla saponinis determinedfrom differencebetweenhydrolyzed andnon-hydrolyzedsaponinsQuillaj...
Future Work Determination of saponin content in plantextracts from Tapeinochuilus annanassae, Syzygiumjambos, Costus spec...
References1. Rodríguez-Tirado, K.; Gavillán-Suárez. University of Puerto Ricoat Cayey, Cayey, P.R. Phytochemical studies o...
References4. Rauter, A.P.; Martins, A.; Lopes, R.; Ferreira, J.; Serralheiro, L.M.;Araújo, M.E.; Borges, C.; Justino, J.; ...
References6. Kumar, A.; Ilavarasan, R.; Jayachandran, T.; Decaraman, N.;Aravindhan, P.; Padmanabhan, N.; Krishnan, M.R.V.P...
Thanks for your attention!
Quantitative analysis ofphytochemicals in antidiabeticplant extractsNatalie Rivera OrtizMentor: Dr. Jannette Gavillán Suár...
of 30

Natalie rivera 28nov_2011

Published on: Mar 3, 2016
Published in: Business      Technology      
Source: www.slideshare.net


Transcripts - Natalie rivera 28nov_2011

  • 1. Quantitative analysis ofphytochemicals in antidiabeticplant extractsNatalie Rivera OrtizMentor: Dr. Jannette Gavillán Suárez, Ph.DDepartment of ChemistryCHEM -4999
  • 2. AcknowledgementsThanks to: Dr. Jannette Gavillán Suárez, Ph.D.
  • 3. What are phytochemicals? They are secondary metabolites that occurnaturally in plants. Molecules responsible for the color and organolepticproperties. They are considered beneficial in treating and/orpreventing chronic diseases.http://www.tjclark.com/phytochemicals//
  • 4. Classification of PhytochemicalsAlkaloidsNicotineCardiac glycosidesOleandrin
  • 5. Classification of PhytochemicalsSaponinsSolanineTerpenoidsUrsolic AcidIsoprene
  • 6. Classification of PhytochemicalsPhenolsFlavonoidsQuercetin
  • 7. Previous Studies Qualitative experiments have demonstrated thepresence of flavonoids, terpenoids, saponins, tanninsand cardiac glycosides in our plant extracts.1 Phenolic compounds protect against oxidativestress and degenerative diseases.2 Oleanolic acid (saponin) has been reported to havehypoglycemic activity.3 In vivo antidiabetic activity of several plant extractshas been correlated with their total phenol andflavonoid content.4, 5
  • 8. Previous Studies Alkaloids, glycosides, carbohydrates, and steroidshave demonstrated activity consistent with theirpossible use in treatment of type-2 diabetes.6Terpenoids have been shown to decrease bloodglucose levels in animal studies.6 Aqueous extract of Acacia arabica (100 μg/mL)stimulated an increase of 228% in insulin release fromrat pancreatic β-cells compared to the control (5.6 mMglucose).7
  • 9. Goal and ObjectivesObjectives:Quantify flavonoids, total phenolic compoundsand saponins in plant extracts from Tapeinochuilusannanassae, Syzygium jambos, Costus speciosus, andTradescantia spathacea.Goal:Quantify possible biomarkers of antidiabeticactivity of plant extracts from several genera.
  • 10. Relevance of this Research Quantification of phytochemicals will be keyin identifying biomarkers of antidiabetic activityof these plants. For the first time, biological activities will becorrelated with phytochemicals’ concentrations.
  • 11. MethodologyFlavonoidsQuercetinsolution(0.27mg/mL)/plantextracts , 200μL of AlCl310%and 200 μLof CH3COONa1MQuercetinCalibrationCurveMeasureabsorbance at415 nm anddetermineconcentration
  • 12. MethodologyTotal Phenolic ContentQuercetinsolution (3.38mg/mL )/plantextracts, 600 μLof Lowryreactive C and200 μL of Lowryreactive EQuercetinCalibrationCurveMeasureabsorbance at760 nm anddetermineconcentration.
  • 13. Results - Flavonoidsy = 34.8x - 0.055R² = 0.9940.00000.10000.20000.30000.40000.50000.60000.70000.80000.00E+00 5.00E-03 1.00E-02 1.50E-02 2.00E-02 2.50E-02Absorbance(415nm)[Quercetin] (mg/mL)Calibration curve for determination offlavonoids in antidiabetic plant extracts
  • 14. Results - FlavonoidsConcentration of plantextract in assay:C1V1=C2V2C2 = C1V1/V2C2 = (0.100mL)(0.04094 g/mL)(5.0000 mL)C2 = 0.000819 g/mL DWFlavonoid content in plantextract:y = mx+bx = y-b/mx = 0.7832-0.00231.9x = 0.0244 mg/mL QECalculations
  • 15. CalculationsResults - FlavonoidsFlavonoid content by dry weight:(0.0224 mg/mL QE)(1mL) = 29.8 mg QE/g DWFlavonoid content by dry weight:(0.0224 mg/mL QE)(1mL) = 29.8 mg QE/g DW0.000819 g DW
  • 16. Results - FlavonoidsPlant Concentrationof flavonoids(± SD mgQE/ g DW)T. spathacea 13.7 ± 0.7C. speciosus 15.8 ± 0.9S. jambos 24.3 ± 1.3T. anannassae 29.8 ± 0.00 5 10 15 20 25 30 35S. jambosT. anannassaeC. speciosusT. spathacea[Flavonoid] (mgQE/g DW)PlantFlavonoid content in antidiabeticplants
  • 17. Results – Total Phenolsy = 4.63x + 0.16R² = 0.9900.00000.10000.20000.30000.40000.50000.60000.70000.80000.90001.00000.00 0.02 0.04 0.06 0.08 0.10 0.12 0.14 0.16 0.18Absorbance(760nm)Quercetin concentration (mg/mL)Calibration curve for determination of totalphenolic content
  • 18. Results – Total PhenolsConcentration ofmethanolic extract inassay:C1V1=C2V2C2 = C1V1/V2C2 = (0.150μL)(3.00 mg/mL)(1500 μL)C2 = 0.300 mg/mL extractTotal phenolic content inmethanolic extract:y = mx+bx = y-b/mx = 0.5603-0.164.63x = 0.085 mg/mL QECalculations
  • 19. Results – Total PhenolsFlavonoid content in methanolic extract:(0.085 mg/mL QE)(1mL) = 0.29 mg QE/mg extract0.300 mg extractFlavonoid content in plant:(0.29 mg QE)(1 mg extract)(1.19 g extract) = 95.33 mg QE/g DW(1 mg extract)(1x10-3g extract)(3.62 g DW)(95.33mg QE)(3.62 g DW) = 7.6 mg QE/ g fresh leaves(1 g DW)(45.66 g fresh leaves)Calculations
  • 20. Results – Total PhenolsAqueousextractTotalPhenolicContent(± SD mgQE)T. spathacea 0.26 ± 0.01C. speciosus 0.35 ± 0.01S. jambos 0.45 ± 0.09T. anannassae 0.14 ± 0.02MethanolicextractTotalPhenolicContent(± SD mgQE)T. spathacea 0.052 ± 0.002C. speciosus 0.060 ± 0.002S. jambos 0.18 ± 0.04T. anannassae 0.034 ± 0.005
  • 21. Results – Total Phenols0 5 10 15S. jambosT.anannassaeC. speciosusT. spathaceaTotal Phenolic Content (mg QE/mg extract)PlantTotal Phenolic Content in Aqueous andMethanolic Extractsaqueousmethanolic
  • 22. Current WorkConcentratedsaponins areextracted twicewith equalvolumes ofn-butanolDNS MethodQuantitativedeterminationof glucose.Glucose (0.4%)and 500 μL ofDNS solution.Calibrationcurve of D-glucose(540 nm)3,5-dinitrosalicylicacidD-glucose 3-amino-5-nitrosalicylic acidD-gluconicacidReaction:
  • 23. Current Worky = 7.390x - 0.003R² = 1.0000.00000.20000.40000.60000.80001.00001.20001.40000.000 0.020 0.040 0.060 0.080 0.100 0.120 0.140 0.160 0.180AbsorbanceConcentration (mg/mL)D-glucose calibration curve for determination of saponins
  • 24. Current WorkSugar content inQuijalla saponinis determinedfrom differencebetweenhydrolyzed andnon-hydrolyzedsaponinsQuillaja saponinSugar content in Quillajasaponin solutions[Quillaja saponin] (mg/mL)[Glucose](mg/mL)
  • 25. Future Work Determination of saponin content in plantextracts from Tapeinochuilus annanassae, Syzygiumjambos, Costus speciosus, and Tradescantia spathacea.Determination of alkaloid content in plantextracts.
  • 26. References1. Rodríguez-Tirado, K.; Gavillán-Suárez. University of Puerto Ricoat Cayey, Cayey, P.R. Phytochemical studies of medicinalplants from several genera with antidiabetic properties.Unpublished work, 2011.2. Han, X.; Shen, T.; Lou, H. Dietary polyphenols and Their Biolo-gical Significance. Int.J.Mol.Sci. [Online] 2008, 8, 950-988.http://www.mdpi.org// (accessed March 8, 2011).3. Güçlü-Ünstündağ, Ö.; Mazza, G. Saponins: properties, applica-tions and processing. Crit. Rev. Food Sci. Nutr. [Online] 2007,47, 231-258. http://www.redorbit.com// (accessed July 14,2011).
  • 27. References4. Rauter, A.P.; Martins, A.; Lopes, R.; Ferreira, J.; Serralheiro, L.M.;Araújo, M.E.; Borges, C.; Justino, J.; Silva, F.V.; Goulart, M.;Thomas-Oates, J.; Rodrigues, J.A.; Edwards, E.; Noronha, J.P.;Pinto, R.; Mota-Filipe, H. Bioactivity studies and chemicalprofile of the antidiabetic plant Genista tenera. J.Ethnopharmacol. [Online] 2009, 122, 384-393. Science Direct.http://www. sciencedirect.com(accessed March 8, 2011).5. Aslan, M.; Deliorman Orhan, D.; Orhan, N.; Sezik, E.; Yesilada, E. Invivo antidiabetic and antioxidant potential of H elichrysumplicatumssp. plicatum capitulums in streptozotocin-induced-diabetic rats. J. Ethnopharmacol. [Online] 2007, 109, 54-59.Science Direct. http://www. sciencedirect.com(accessedNovember 20, 2011).
  • 28. References6. Kumar, A.; Ilavarasan, R.; Jayachandran, T.; Decaraman, N.;Aravindhan, P.; Padmanabhan, N.; Krishnan, M.R.V.Phytochemicals Investigation on a Tropical Plant, Syzygiumcumini fromKattuppalayam, Erode District, Tamil Nadu, SouthIndia. Pak. J. Nutr. [Online] 2009, 8, 1, 83-85.http://www.pjbs.org/pjnonline// (accessed March 8, 2011).7. Kaur, L.; Han, K.S.; Bains, K.; Singh, H. Indian culinary plantsenhance glucose-induced insulin secretion and glucoseconsumption in INS-1 b-cells and 3T3-L1 adipocytes. FoodChem. [Online] 2011, 29, 1120-1125. Science Direct.http://www. sciencedirect.com(accessed November 20,2011).
  • 29. Thanks for your attention!
  • 30. Quantitative analysis ofphytochemicals in antidiabeticplant extractsNatalie Rivera OrtizMentor: Dr. Jannette Gavillán Suárez, Ph.DDepartment of ChemistryCHEM -4999

Related Documents