Lignan Concentration

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.

Results and Discussion

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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.

Figure 1: HPLC analysis of 10.0 μL of Lignan concentrate in 1.0 mL of H2O. 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.

The purification and concentration of HMR Lignans for Life is an accessible way of obtaining these hydroxymatairesinol for experimental use. The relative purity and ease of which this substance was concentrated leaves avenues of utilization in terms of future experiments of lignan oxidation and degradation. The HPLC Chromatogram of the concentrated HMR Lignans for Life sample indicates that this material is able to be concentrated and can be used in future experiments