Lignan Concentration
Materials and Methods
Lignans for Life Hydroxymatairesinol (HMR) Lignan was obtained from Amazon.com (Seattle, WA, USA), and 1,4-Dioxane and EtOH 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 28℃ 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 and spun overnight to remove the solvent. The pelleted solute in each Eppendorf tube was then resolubilized using 1 mL of EtOH.
The Lignan concentrate (10 μL) in 1 mL of H2O was ran on the HPLC at absorbance wavelength of 270 nm and 28℃ 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.