Presentation Title

The Effect of Hierba Manayupa Extractions on HeLa Cells and the Identification of Chemical Composition via GC-MS

Faculty Mentor

Dr. Sylvine Deprele, Dr. Luiza Nogaj

Start Date

17-11-2018 8:30 AM

End Date

17-11-2018 8:45 AM

Location

C302

Session

Oral 1

Type of Presentation

Oral Talk

Subject Area

health_nutrition_clinical_science

Abstract

Natural products contain beneficial compounds and traces of possible toxic byproducts that are detrimental to one’s health. Polyphenols and Polycyclic aromatic hydrocarbons (PAHs) exemplify these good and bad compounds that are actively competing with one another due to their opposite effects in regards to human health. Polyphenols are naturally occurring organic compounds that are known antioxidants that reduce a tumors’ size and growth. PAHs are naturally occurring chemicals that are known carcinogens, mutagens, and teratogens. This study focused on observing the effects Hierba Manayupa extracts had on the cell viability of HeLa cells via MTT Assay and identifying its chemical composition using Gas Chromatography Mass Spectrometry (GC-MS) analysis. The tea leaves were extracted using a Soxhlet apparatus in the solvent systems: water, dichloromethane, and ethanol/acetone (1:1) followed by a series of liquid-liquid extractions that used chloroform, ethyl acetate, (90 mL) sodium bicarbonate, and brine. The samples were then filtered, evaporated and re-suspended in the appropriate solvents. The extracts were analyzed via the GC-MS to identify the active compounds found within Hierba Manuyapa. The following PAHs was detected in high concentrations within the water and dichloromethane extracts, respectively: Naphthalene 670.4 and 674.4 . Polyphenols and possible derivatives were detected, but further analysis is being conducted. MTT assays found that all three Hierba Manayupa samples had great effects on HeLa cells at 5 µL and 10 µL concentrations, for they caused a distinctive decrease in cell viability. Cytotoxicity assays examined cell toxicity and concluded that all three extracts were nontoxic to the cell membrane at the 5, 10, and 20 µL concentrations. Fractioning of the herbal extracts via column chromatography is currently being conducted to further separate the identified compounds. Future directions include further research on the observed polyphenols and derivatives of the herb to ensure they possess true antioxidant properties.

Summary of research results to be presented

In our research, various cell viability experiments using MTT assays were conducted on Hierba Manayupa extracts. Therefore, it was concluded that at a 1:10 dilution, all three Hierba Manayupa solvent samples decreased cell viability at the 5 µL and 10 µL concentrations and at a 1:20 dilution, the dichloromethane solvent sample efficiently decreased cell viability at all three concentrations. The herbal extracts were also analyzed via Gas Chromatography Mass Spectrometry (GC-MS) in which PAHs, such as Naphthalene, were detected in high concentrations within the water and dichloromethane extracts at 670.4 µg/µL (water) and 674.4 µg/µL(dichloromethane). Possible polyphenols and their derivatives were also detected in the extracts. Cell viability experiments with a cytotoxicity assay were also conducted and it was concluded that all three extracts were nontoxic to the cell membrane at the 5, 10, and 20 µL concentrations.

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Nov 17th, 8:30 AM Nov 17th, 8:45 AM

The Effect of Hierba Manayupa Extractions on HeLa Cells and the Identification of Chemical Composition via GC-MS

C302

Natural products contain beneficial compounds and traces of possible toxic byproducts that are detrimental to one’s health. Polyphenols and Polycyclic aromatic hydrocarbons (PAHs) exemplify these good and bad compounds that are actively competing with one another due to their opposite effects in regards to human health. Polyphenols are naturally occurring organic compounds that are known antioxidants that reduce a tumors’ size and growth. PAHs are naturally occurring chemicals that are known carcinogens, mutagens, and teratogens. This study focused on observing the effects Hierba Manayupa extracts had on the cell viability of HeLa cells via MTT Assay and identifying its chemical composition using Gas Chromatography Mass Spectrometry (GC-MS) analysis. The tea leaves were extracted using a Soxhlet apparatus in the solvent systems: water, dichloromethane, and ethanol/acetone (1:1) followed by a series of liquid-liquid extractions that used chloroform, ethyl acetate, (90 mL) sodium bicarbonate, and brine. The samples were then filtered, evaporated and re-suspended in the appropriate solvents. The extracts were analyzed via the GC-MS to identify the active compounds found within Hierba Manuyapa. The following PAHs was detected in high concentrations within the water and dichloromethane extracts, respectively: Naphthalene 670.4 and 674.4 . Polyphenols and possible derivatives were detected, but further analysis is being conducted. MTT assays found that all three Hierba Manayupa samples had great effects on HeLa cells at 5 µL and 10 µL concentrations, for they caused a distinctive decrease in cell viability. Cytotoxicity assays examined cell toxicity and concluded that all three extracts were nontoxic to the cell membrane at the 5, 10, and 20 µL concentrations. Fractioning of the herbal extracts via column chromatography is currently being conducted to further separate the identified compounds. Future directions include further research on the observed polyphenols and derivatives of the herb to ensure they possess true antioxidant properties.