User:Moira M. Esson/Notebook/CHEM-571/2013/10/01: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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Figure 1. Corrected Absorbance Spectra of preliminary kinetics | Figure 1. Corrected Absorbance Spectra of preliminary kinetics | ||
<br> | <br> | ||
[[Image:HRP diluteluminol h2o2 zem.png]] | [[Image:HRP diluteluminol h2o2 zem.png|500px]] | ||
<br> | <br> | ||
*The H<sub>2</sub>O<sub>2</sub> stock solution appears to be too dilute to produce a spectrum. | |||
*A 1/20 dilution of luminol was used in order to produce an absorbance peak with a value below 1. | |||
==Kinetics of luminol oxidized by HRP== | |||
<br> | |||
'''General Protocol:''' | |||
#Add the desired HRP/Luminol stock solution to the stop flow mixer. | |||
#Add H<sub>2</sub>O<sub>2</sub> stock solution to the stop flow mixer. | |||
#Equilibrate mixer tubes with sample | |||
#*Note: For the steps described above, no air bubbles can be present in the sample. Air bubbles will affect the kinetics data(oxygen will act as an oxidant and produce unreliable data) | |||
#Initiate Mixing | |||
#Take a absorbance spectrum of the sample every 30 seconds for 10 minutes in order to monitor the reaction | |||
#Using the luminol and 3-aminophthalic acid spectra as endpoints, determine the kinetics of the 3-aminophthalic acid synthesis | |||
<br> | |||
'''Data collection''': | |||
*The 1/20 dilution of luminol was used for the reaction mixture | |||
*A 1/20 dilution of hydrogen peroxide stock solution was used based on other groups findings | |||
*Stock solution of HRP was used | |||
*The sample which was run was completely oxidized upon mixture of the reaction. This may be due to the ratio of luminol to the concentrated HRP. Another group's data will be analyzed and the kinetics procedure will be repeated again tomorrow. | |||
'''Results''': | |||
The below results were obtained from [[User:Trisha I. Ibeh/Notebook/Trisha Notebook/2013/10/01|Trisha's group]] | |||
*A diluted solution of luminol with a concentration of .126mM was used with the stock solutions of HRP and hydrogen peroxide | |||
<br> | |||
'''Figure 2. Absorbance spectra of the oxidation of luminol by HRP focusing on the 350nm region''' | |||
[[Image:Kineticsmonitoringzem10012013.png|500px]] | |||
<br> | |||
*The wavelength maximum occurs at 348.99nm and experiences a growth in signal. This indicates the production of 3-aminophthalic acid. | |||
<br> | |||
'''Figure 3. Absorption spectra of the production of 3-aminopthalate by the oxidaiton of luminol by HRP focusing on the 420nm region''' | |||
[[Image:Kinematicsincreaseprdtzem10012013.png|500px]] | |||
<br> | |||
*The wavelength maximum occurs at 415.42nm. During the course of the ten minutes, a signal reduction occurred, indicating the oxidation of luminol. | |||
Latest revision as of 23:24, 26 September 2017
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Objectives
UV-visStock solutions prepared before class:
Samples Run:
Kinetics of luminol oxidized by HRP
Results: The below results were obtained from Trisha's group
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