Presentation Title

The Effect of Hierba Santa Extractions on HeLa Cells and the Identification of its Chemical Composition Via GC/MS

Faculty Mentor

Sylvine Deprèle PhD, Luiza Nogaj PhD

Start Date

17-11-2018 8:30 AM

End Date

17-11-2018 8:45 AM

Location

C301

Session

Oral 1

Type of Presentation

Oral Talk

Subject Area

health_nutrition_clinical_science

Abstract

Polyphenols are compounds that act as antioxidants and display anti-carcinogenic properties. Polycyclic aromatic hydrocarbons (PAHs) are compounds resulting from incomplete combustion and exposure to them can lead to harmful repercussions since they are carcinogenic, mutagenic and teratogenic. Since polyphenols and PAHs have opposite effects in human health, their intake is critical in determining the benefit of their combined effects. Previous research tested six Peruvian teas for effectiveness in decreasing the cell viability of HeLa cells and found Hierba Santa in teas displaying positive results. The herb is known to have antioxidant properties thus, the objective tested Hierba Santa extracts to observe their effects on HeLa cells via MTT and cytotoxicity assays, and analyze their chemical composition using the GC/MS to understand its potential role in cell viability. Soxhlet extractions were conducted with three solvent systems: dichloromethane, water, and ethanol/acetone (1:1). Liquid/liquid extractions with chloroform, ethyl acetate (90mL), sodium bicarbonate, and brine were performed to remove impurities from the herbal extracts. The solvents were then evaporated and the extracts were resuspended in appropriate solvents for GC/MS analysis and cell work. MTT assays showed that the Hierba Santa dichloromethane and water extracts decreased the cell viability of HeLa cells at the 5, 10, and 20 uL concentrations. The observed cytotoxicity assay results demonstrated that at all three Hierba Santa extracts exhibited an increase in membrane toxicity at the 5, 10, and 20 uL concentrations. Different programs on the GC/MS identified polyphenol derivatives and PAHs within the herbal extracts. Identified PAHs include: naphthalene, benz(a)anthr, benz(b)fluor, benz(k)fluor, and benz(a)pyre. We are currently fractioning Hierba Santa extracts based on their polarity and conducting MTT and cytotoxicity assays on the fractions. Future directions include classifying identified polyphenol derivatives.

Summary of research results to be presented

MTT assays showed that the Hierba Santa dichloromethane and water extracts decreased the cell viability of HeLa cells at the 5, 10, and 20 uL concentrations. The observed cytotoxicity assay results demonstrated that at all three Hierba Santa extracts exhibited an increase in membrane toxicity at the 5, 10, and 20 uL concentrations. Different programs on the GC/MS identified polyphenol derivatives and PAHs within the herbal extracts. Identified PAHs include: naphthalene, benz(a)anthr, benz(b)fluor, benz(k)fluor, and benz(a)pyre.

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

The Effect of Hierba Santa Extractions on HeLa Cells and the Identification of its Chemical Composition Via GC/MS

C301

Polyphenols are compounds that act as antioxidants and display anti-carcinogenic properties. Polycyclic aromatic hydrocarbons (PAHs) are compounds resulting from incomplete combustion and exposure to them can lead to harmful repercussions since they are carcinogenic, mutagenic and teratogenic. Since polyphenols and PAHs have opposite effects in human health, their intake is critical in determining the benefit of their combined effects. Previous research tested six Peruvian teas for effectiveness in decreasing the cell viability of HeLa cells and found Hierba Santa in teas displaying positive results. The herb is known to have antioxidant properties thus, the objective tested Hierba Santa extracts to observe their effects on HeLa cells via MTT and cytotoxicity assays, and analyze their chemical composition using the GC/MS to understand its potential role in cell viability. Soxhlet extractions were conducted with three solvent systems: dichloromethane, water, and ethanol/acetone (1:1). Liquid/liquid extractions with chloroform, ethyl acetate (90mL), sodium bicarbonate, and brine were performed to remove impurities from the herbal extracts. The solvents were then evaporated and the extracts were resuspended in appropriate solvents for GC/MS analysis and cell work. MTT assays showed that the Hierba Santa dichloromethane and water extracts decreased the cell viability of HeLa cells at the 5, 10, and 20 uL concentrations. The observed cytotoxicity assay results demonstrated that at all three Hierba Santa extracts exhibited an increase in membrane toxicity at the 5, 10, and 20 uL concentrations. Different programs on the GC/MS identified polyphenol derivatives and PAHs within the herbal extracts. Identified PAHs include: naphthalene, benz(a)anthr, benz(b)fluor, benz(k)fluor, and benz(a)pyre. We are currently fractioning Hierba Santa extracts based on their polarity and conducting MTT and cytotoxicity assays on the fractions. Future directions include classifying identified polyphenol derivatives.