Difference between revisions of "Lignan Concentration"
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===Materials and Methods=== | ===Materials and Methods=== | ||
| − | Lignans for Life Hydroxymatairesinol (HMR) Lignan was obtained from Amazon.com (Seattle, WA, USA), and 1,4-Dioxane and | + | Lignans for Life Hydroxymatairesinol (HMR) Lignan was obtained from Amazon.com (Seattle, WA, USA), and 1,4-Dioxane and ethanol were obtained from Sigma Aldrich (St. Louis, MO, USA). |
| − | Solutions of 2 HMR Lignan capsules + 20 mL of 1,4-Dioxane and 4 HMR Lignan capsules + 40 mL of 1,4 Dioxane were prepared. These solutions were filtered and analyzed using the HPLC at absorbance wavelength of 270 nm and | + | Solutions of 2 HMR Lignan capsules + 20.0 mL of 1,4-Dioxane and 4 HMR Lignan capsules + 40.0 mL of 1,4 Dioxane were prepared. These solutions were filtered and analyzed using the HPLC at absorbance wavelength of 270 nm and 28°C using an acetonitrile (ACN) and 0.1% TFA gradient for 30 minutes. The first 15 minutes were run at 100% 0.1% TFA, 15-25 minutes at 100% ACN, and 25-30 minutes at 100% 0.1% TFA. |
| − | The solution of 4 HMR Lignan capsules in 40 mL of 1,4-Dioxane was used to fill ten 1.5 mL Eppendorf tubes with 1.0 mL of solution. These Eppendorf tubes were placed in a Savant Speedvac SC110 Concentrator (Holbrook, NY, USA) and spun overnight to remove the solvent. The pelleted solute in each Eppendorf tube was then resolubilized using 1 mL of EtOH. | + | The solution of 4 HMR Lignan capsules in 40 mL of 1,4-Dioxane was used to fill ten 1.5 mL Eppendorf tubes with 1.0 mL of solution. These Eppendorf tubes were placed in a Savant Speedvac SC110 Concentrator (Holbrook, NY, USA) and spun overnight to remove the solvent. The pelleted solute in each Eppendorf tube was then resolubilized using 1.0 mL of EtOH. |
| − | The Lignan concentrate (10 μL) in 1 mL of H<sub>2</sub>O was ran on the HPLC at absorbance wavelength of 270 nm and | + | The Lignan concentrate (10 μL) in 1.0 mL of H<sub>2</sub>O was ran on the HPLC at absorbance wavelength of 270 nm and 28°C using an acetonitrile (ACN) and 0.1% TFA gradient for 30 minutes. The first 15 minutes were run at 100% 0.1% TFA, 15-25 minutes at 100% ACN, and 25-30 minutes at 100% 0.1% TFA. |
===Results and Discussion=== | ===Results and Discussion=== | ||
| − | [[File:2.20.4.40 Wiki layout.png|400px|thumb|left|Figure 1: HPLC analysis of varying concentrations of 2 HMR Lignan Capsules in 20 mL of 1,4-Dioxane and 4 HMR Lignan Capsules in 40 mL of 1,4-Dioxane.]] | + | [[File:2.20.4.40 Wiki layout.png|400px|thumb|left|Figure 1: HPLC analysis of varying concentrations of 2 HMR Lignan Capsules in 20.0 mL of 1,4-Dioxane and 4 HMR Lignan Capsules in 40.0 mL of 1,4-Dioxane. The HPLC was run at 28.0°C with a gradient elution at 0-15 min: 100% 0.1% TFA, 0% acetonitrile, 15-25 min: 0% 0.1% TFA, 100% acetonitrile, 25-30 min: 100% 0.1% TFA, 0% acetonitrile. Trials were run once.]] |
| − | [[File:1uL in 1mL h20.png|400px|thumb|right|Figure 1: HPLC analysis of 10 μL of Lignan concentrate in 1 mL of H<sub>2</sub>O]] | + | [[File:1uL in 1mL h20.png|400px|thumb|right|Figure 1: HPLC analysis of 10.0 μL of Lignan concentrate in 1.0 mL of H<sub>2</sub>O. The HPLC was run at 28.0°C with a gradient elution at 0-15 min: 100% 0.1% TFA, 0% acetonitrile, 15-25 min: 0% 0.1% TFA, 100% acetonitrile, 25-30 min: 100% 0.1% TFA, 0% acetonitrile. Trials were run once.]] |
Revision as of 18:13, 6 April 2021
Materials and Methods
Lignans for Life Hydroxymatairesinol (HMR) Lignan was obtained from Amazon.com (Seattle, WA, USA), and 1,4-Dioxane and ethanol were obtained from Sigma Aldrich (St. Louis, MO, USA).
Solutions of 2 HMR Lignan capsules + 20.0 mL of 1,4-Dioxane and 4 HMR Lignan capsules + 40.0 mL of 1,4 Dioxane were prepared. These solutions were filtered and analyzed using the HPLC at absorbance wavelength of 270 nm and 28°C using an acetonitrile (ACN) and 0.1% TFA gradient for 30 minutes. The first 15 minutes were run at 100% 0.1% TFA, 15-25 minutes at 100% ACN, and 25-30 minutes at 100% 0.1% TFA.
The solution of 4 HMR Lignan capsules in 40 mL of 1,4-Dioxane was used to fill ten 1.5 mL Eppendorf tubes with 1.0 mL of solution. These Eppendorf tubes were placed in a Savant Speedvac SC110 Concentrator (Holbrook, NY, USA) and spun overnight to remove the solvent. The pelleted solute in each Eppendorf tube was then resolubilized using 1.0 mL of EtOH.
The Lignan concentrate (10 μL) in 1.0 mL of H2O was ran on the HPLC at absorbance wavelength of 270 nm and 28°C using an acetonitrile (ACN) and 0.1% TFA gradient for 30 minutes. The first 15 minutes were run at 100% 0.1% TFA, 15-25 minutes at 100% ACN, and 25-30 minutes at 100% 0.1% TFA.
