Lignan Concentration - Revision history

Tcoffman: /* Results and Discussion */

2021-05-05T14:43:03Z

Results and Discussion

Tcoffman: /* Results and Discussion */

2021-05-05T14:41:38Z

Results and Discussion

Tcoffman: /* Results and Discussion */

2021-05-05T13:49:58Z

Results and Discussion

Tcoffman: /* Results and Discussion */

2021-05-05T13:49:32Z

Results and Discussion

Tcoffman: /* Results and Discussion */

2021-05-04T21:41:23Z

Results and Discussion

Tcoffman: /* Results and Discussion */

2021-05-04T21:39:44Z

Results and Discussion

Tcoffman: /* Materials and Methods */

2021-05-04T21:09:53Z

Materials and Methods

Tcoffman: /* Results and Discussion */

2021-05-04T21:06:34Z

Results and Discussion

Tcoffman: /* Results and Discussion */

2021-05-04T21:03:21Z

Results and Discussion

Tcoffman: /* Materials and Methods */

2021-05-04T20:58:22Z

Materials and Methods

@@ Line 11: / Line 11: @@
 {|
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-||[[File:2.20.4.40 Wiki layout.png|450px|thumb|Figure 1: Offset HPLC analysis 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 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.]] || [[File:1uL in 1mL h20.png|450px|thumb|Figure 1: HPLC analysis of 10.0 μL of Lignan concentrate in 1.0 mL of H<sub>2</sub>O. 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.]]
+||[[File:lignanconcentration1.png|450px|thumb|Figure 1: Offset HPLC analysis 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 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.]] || [[File:1uL in 1mL h20.png|450px|thumb|Figure 1: HPLC analysis of 10.0 μL of Lignan concentrate in 1.0 mL of H<sub>2</sub>O. 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.]]
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@@ Line 11: / Line 11: @@
 {|
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-||[[File:220440.png|450px|thumb|Figure 1: Offset HPLC analysis 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 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.]] || [[File:1uL in 1mL h20.png|450px|thumb|Figure 1: HPLC analysis of 10.0 μL of Lignan concentrate in 1.0 mL of H<sub>2</sub>O. 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.]]
+||[[File:2.20.4.40 Wiki layout.png|450px|thumb|Figure 1: Offset HPLC analysis 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 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.]] || [[File:1uL in 1mL h20.png|450px|thumb|Figure 1: HPLC analysis of 10.0 μL of Lignan concentrate in 1.0 mL of H<sub>2</sub>O. 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.]]
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@@ Line 11: / Line 11: @@
 {|
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-||[[File:220440.png|450px|thumb|Figure 1: Offset 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 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.]] || [[File:1uL in 1mL h20.png|450px|thumb|Figure 1: HPLC analysis of 10.0 μL of Lignan concentrate in 1.0 mL of H<sub>2</sub>O. 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.]]
+||[[File:220440.png|450px|thumb|Figure 1: Offset HPLC analysis 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 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.]] || [[File:1uL in 1mL h20.png|450px|thumb|Figure 1: HPLC analysis of 10.0 μL of Lignan concentrate in 1.0 mL of H<sub>2</sub>O. 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.]]
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@@ Line 11: / Line 11: @@
 {|
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-||[[File:2.20.4.40 Wiki layout.png|450px|thumb|Figure 1: Offset 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 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.]] || [[File:1uL in 1mL h20.png|450px|thumb|Figure 1: HPLC analysis of 10.0 μL of Lignan concentrate in 1.0 mL of H<sub>2</sub>O. 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.]]
+||[[File:220440.png|450px|thumb|Figure 1: Offset 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 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.]] || [[File:1uL in 1mL h20.png|450px|thumb|Figure 1: HPLC analysis of 10.0 μL of Lignan concentrate in 1.0 mL of H<sub>2</sub>O. 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.]]
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@@ Line 11: / Line 11: @@
 {|
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-||[[File:2.20.4.40 Wiki layout.png|450px|thumb|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 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.]] || [[File:1uL in 1mL h20.png|450px|thumb|Figure 1: HPLC analysis of 10.0 μL of Lignan concentrate in 1.0 mL of H<sub>2</sub>O. 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.]]
+||[[File:2.20.4.40 Wiki layout.png|450px|thumb|Figure 1: Offset 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 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.]] || [[File:1uL in 1mL h20.png|450px|thumb|Figure 1: HPLC analysis of 10.0 μL of Lignan concentrate in 1.0 mL of H<sub>2</sub>O. 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.]]
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@@ Line 10: / Line 10: @@
 ===Results and Discussion===
 {|
-[[File:2.20.4.40 Wiki layout.png|400px|thumb|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 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.]]
+|-
-||
+||[[File:2.20.4.40 Wiki layout.png|450px|thumb|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 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.]] || [[File:1uL in 1mL h20.png|450px|thumb|Figure 1: HPLC analysis of 10.0 μL of Lignan concentrate in 1.0 mL of H<sub>2</sub>O. 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.]]
-[[File:1uL in 1mL h20.png|400px|thumb|Figure 1: HPLC analysis of 10.0 μL of Lignan concentrate in 1.0 mL of H<sub>2</sub>O. 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 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.

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 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 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 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.
+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 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.
 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 H<sub>2</sub>O was ran on 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 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 Lignan concentrate (10 μL) in 1.0 mL of H<sub>2</sub>O was ran on 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===

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 ===Results and Discussion===
 {|
-[[File:2.20.4.40 Wiki layout.png|400px|thumb|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:2.20.4.40 Wiki layout.png|400px|thumb|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 sample was run through an Agilent 1100 HPLC with a C18 column with a length of 4.6 x 100 mm and a particle size of 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.]]
 ||
-[[File:1uL in 1mL h20.png|400px|thumb|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.]]
+[[File:1uL in 1mL h20.png|400px|thumb|Figure 1: HPLC analysis of 10.0 μL of Lignan concentrate in 1.0 mL of H<sub>2</sub>O. The sample was run through an Agilent 1100 HPLC with a C18 column with a length of 4.6 x 100 mm and a particle size of 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 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.