Lignan Purification

Lignan Purification

Materials and Methods

Lignans for Life Hydroxymatairesinol (HMR) Lignan was obtained from Amazon.com (Seattle, WA, USA) and ethanol was obtained from Sigma Aldrich (St. Louis, MO, USA).

Solutions of 1 HMR Lignan capsule (0.169 g), 5 HMR Lignan capsules (0.785 g), and 100.0 mL of EtOH and 10 HMR Lignan capsules (1.629 g) and 100.0 mL of EtOH were prepared. These solutions were filtered and analyzed using an Agilent 1100 Series HPLC with a C18 column (4.16 x 100 mm length and 3.5 μm particle size) at absorbance wavelength of 270 nm and temperature of 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

Figure 1: Offset HPLC analysis of 1, 5, and 10 HMR Lignan capsules in 100.0 mL of ethanol. The sample was run through an Agilent 1100 HPLC with a C18 column (length: 4.6 x 100 mm and particle size: 3.5 μm) 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 of lignan using an easily accessible source such as Lignans for Life’s HMR Lignans seems to be a promising method of gaining lignan for experimental use. Purification of these HMR lignan capsules produced a chromatogram that has a single peak, indicating that our sample is a relatively pure compound with little pollution (Figure 1.). As the absorption of each peak increases with higher mass of solute, we see that the amount of uncharacterized compound in the HMR Lignan capsules is relatively homogeneous across the samples. Because we cannot be sure that these HMR Lignan capsules contain hydroxymatairesinol, further characterization steps will need to be taken to ensure that the claims of the manufacturers are accurate.

Future Directions

Because this project has not reached any conclusion as to whether Lignans for Life HMR Lignan truly contains hydroxymatairesinol, further characterization of this powder must be performed. This can be done through nuclear magnetic resonance or through mass spectroscopy. Once a characterization of this unknown is completed, different tests can be performed in order to better understand the biological activity of this specific lignan, including a Kirby-Bauer test. If this lignan is easily isolated, this could also provide material for further lignin/lignan degradation and synthesis work.