N-Acetyl L-Tyrosine

This page depicts the oxidation of N-Acetyl L-Tyrosine (NAT) by Horse Radish Peroxidase (HRP). Each sample of NAT was dissolved in pH 5 phosphate citrate buffer. The method used on the HPLC was "HPA_0604156".

NAT

The Dioxane Conundrum

The following two graphs depict the difference in reaction when dioxane is present and when it is not. The conclusion is that dioxane quenches the oxidation of NAT.

Figure 1: NAT oxidation in solution with dioxane and reacted with H2O2 and HRP 6/5/16. This graph shows the dioxane in the solution inhibiting the reaction completely.

Figure 2: 2mM NAT oxidation without dioxane and reacted with 2mM H2O2 and HRP in a beaker 6/5/16

Beaker Reaction vs. Immobilized Enzyme Bio-Reactor

The following graphs depict the difference in HPLC data when the reaction is conducted in a beaker and when the reaction is ran using the immobilized enzyme technique using a bio-reactor.

Figure 2: 2mM NAT reacted with 2mM H2O2 and HRP in a beaker 6/5/16

Figure 3: 2mM NAT reacted with 1mM H2O2 using the Immobilized Enzyme technique 6/9/16

Concentration Test

These graphs depict the difference in concentration of NAT and H2O2. Increasing the concentration will help to obtain more products.

Figure 4: 2mM NAT oxidation without dioxane and reacted with 0.5mM, 1.0mM, and 2.0mM H2O2 with immobilized HRP at 0.5ml/min 6/29/16

Figure 5: 10mM NAT oxidized with 5, 10, and 20mM H2O2 with immobilized HRP at 0.5ml/min 6/23/16

Flash Chromatography

The objective of flash chromatography is to isolate peaks seen in the HPLC. Running flash chromatography allows for the product to be isolated.

Figure 6: Flash chromatography separation of the 10mM H2O2 in Figure 5 6/28/16

The following graphs show the difference between running the flash system under conditions with wavelength at 276 and 300 with 223 and 276. The first graph is the NAT standard ran through the HPLC. The two flash chromatography graphs show the difference in changing the methods of the flash system.

HPLC chromatogram of 20mM NAT.

20mM NAT ran through flash with the wavelength red and purple being 276 and 300, respectively.

20mM NAT ran through flash with the wavelength red and purple being 223 and 276, respectively.

Antioxidants in Reaction

The following graph depicts the oxidation of 2mM NAT by H2O2 with various concentrations of ascorbic acid in pH 5 buffer.

Figure 7: 2mM NAT oxidation with 1mM H202 with 0.25mM, 0.5mM, 1mM, 2mM ascorbic acid 7/5/16

The following is the oxidation of 2mM NAT by H2O2 with various concentrations of reduced GSH in pH 5 buffer.

Figure 7: 2mM NAT oxidation with 1mM H202 with 0.25mM, 0.5mM, 1mM, 2mM GSH. 7/7/16