Presentation Title

Quantification of Rutin and Chlorogenic acid in Sambucus mexicana extracts using HPLC

Faculty Mentor

P. Matthew Joyner

Start Date

17-11-2018 8:30 AM

End Date

17-11-2018 10:30 AM

Location

CREVELING 46

Session

POSTER 1

Type of Presentation

Poster

Subject Area

biological_agricultural_sciences

Abstract

Inflammation is one of the body’s responses to injury. The Chumash people of Southern California used many plants as medicinal treatments for various types of injuries. One such plant is Sambucus mexicana. In this study, two anti-inflammatory compounds- rutin and chlorogenic acid- found in Sambucas mexicana, were analyzed for their effects on nitric oxide production in RAW 264.7 macrophage cells and to determine if the leaves or flowers of S. mexicana contained a higher concentration of rutin and chlorogenic acid. Extractions were done in a 6:4 methanol:water solvent with rotary evaporation and two vacuum filtration processes using an SPE column and filter paper. The extracts’ concentrations of rutin and chlorogenic acid were analyzed with High Performance Liquid Chromatography using 0.25% phosphoric acid and 100% acetonitrile mobile phases. Data for each sample was acquired over 30 minutes using a low-pressure binary gradient and total pump flow of 0.5mL/min. Average concentrations of rutin in six flower and five leaf extracts was 290.6±105.3 g/mL and 107.8±69.8 g/mL and the average concentration of chlorogenic acid in flower and leaf extracts was 740.6±276.8g/mL and 121.6±81.3 g/mL,suggesting that the amounts of rutin and chlorogenic acid are higher in flowers, however the concentrations of the compounds varied in each extract. Nitric oxide production in RAW 264.7 macrophage cells was tested using the Griess Reagent System, which is based upon a chemical reaction with Sulfanilamide and N-1-napthylethylenediamine dihydroxychloride. Decreases in nitric oxide were detected by measuring the absorbance at 520nm using a plate reader. Rutin and chlorogenic acid decreased nitric oxide production in macrophages. While our results confirm the anti-inflammatory properties of rutin and chlorogenic acid, data also suggests the presence of additional unknown anti-inflammatory compounds in S. mexicana. Future research can be done to identify these unknown compounds.

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Nov 17th, 8:30 AM Nov 17th, 10:30 AM

Quantification of Rutin and Chlorogenic acid in Sambucus mexicana extracts using HPLC

CREVELING 46

Inflammation is one of the body’s responses to injury. The Chumash people of Southern California used many plants as medicinal treatments for various types of injuries. One such plant is Sambucus mexicana. In this study, two anti-inflammatory compounds- rutin and chlorogenic acid- found in Sambucas mexicana, were analyzed for their effects on nitric oxide production in RAW 264.7 macrophage cells and to determine if the leaves or flowers of S. mexicana contained a higher concentration of rutin and chlorogenic acid. Extractions were done in a 6:4 methanol:water solvent with rotary evaporation and two vacuum filtration processes using an SPE column and filter paper. The extracts’ concentrations of rutin and chlorogenic acid were analyzed with High Performance Liquid Chromatography using 0.25% phosphoric acid and 100% acetonitrile mobile phases. Data for each sample was acquired over 30 minutes using a low-pressure binary gradient and total pump flow of 0.5mL/min. Average concentrations of rutin in six flower and five leaf extracts was 290.6±105.3 g/mL and 107.8±69.8 g/mL and the average concentration of chlorogenic acid in flower and leaf extracts was 740.6±276.8g/mL and 121.6±81.3 g/mL,suggesting that the amounts of rutin and chlorogenic acid are higher in flowers, however the concentrations of the compounds varied in each extract. Nitric oxide production in RAW 264.7 macrophage cells was tested using the Griess Reagent System, which is based upon a chemical reaction with Sulfanilamide and N-1-napthylethylenediamine dihydroxychloride. Decreases in nitric oxide were detected by measuring the absorbance at 520nm using a plate reader. Rutin and chlorogenic acid decreased nitric oxide production in macrophages. While our results confirm the anti-inflammatory properties of rutin and chlorogenic acid, data also suggests the presence of additional unknown anti-inflammatory compounds in S. mexicana. Future research can be done to identify these unknown compounds.