User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/10/09 - Revision history

Mary Mendoza: /* Fluorescence Spectrometer Graph */

Mary Mendoza — Fri, 07 Dec 2012 13:56:07 GMT

Fluorescence Spectrometer Graph

←Older revision		Revision as of 13:56, 7 December 2012
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	{\| {{table}}		{\| {{table}}
		+	\| align="center" style="background:#f0f0f0;"\|''''
	\| align="center" style="background:#f0f0f0;"\|''HRP''		\| align="center" style="background:#f0f0f0;"\|''HRP''
	\| align="center" style="background:#f0f0f0;"\|''4-iodophenol''		\| align="center" style="background:#f0f0f0;"\|''4-iodophenol''
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	\| align="center" style="background:#f0f0f0;"\|''H<sub>2</sub>O''		\| align="center" style="background:#f0f0f0;"\|''H<sub>2</sub>O''
	\|-		\|-
-	\| 30 μL\|\|54 μL\|\|900 μL\|\|176 μL\|\|1.8 mL	+	\| Molarity of Stock Solutions\|\|2.3 μM\|\|18 mM\|\|10 mM\|\|1.7 mM\|\|neutral
		+	\|-
		+	\| Volume\|\|30 μL\|\|54 μL\|\|900 μL\|\|176 μL\|\|1.8 mL
	\|-		\|-

Mary Mendoza: /* Fluorescence Spectrometer Graph */

Mary Mendoza — Fri, 26 Oct 2012 07:06:55 GMT

Fluorescence Spectrometer Graph

←Older revision		Revision as of 07:06, 26 October 2012
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	{\| {{table}}		{\| {{table}}
-	\| align="center" style="background:#f0f0f0;"\|''~~sample~~''	+	\| align="center" style="background:#f0f0f0;"\|''HRP''
	\| align="center" style="background:#f0f0f0;"\|''4-iodophenol''		\| align="center" style="background:#f0f0f0;"\|''4-iodophenol''
	\| align="center" style="background:#f0f0f0;"\|''luminol''		\| align="center" style="background:#f0f0f0;"\|''luminol''
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	\| align="center" style="background:#f0f0f0;"\|''H<sub>2</sub>O''		\| align="center" style="background:#f0f0f0;"\|''H<sub>2</sub>O''
	\|-		\|-
-	\| 1\|\|30 μL\|\|900 μL\|\|176 μL\|\|1.8 mL	+	\| 30 μL\|\|54 μL\|\|900 μL\|\|176 μL\|\|1.8 mL
	\|-		\|-

Mary Mendoza: /* Fluorescence Spectrometer Graph */

Mary Mendoza — Fri, 26 Oct 2012 07:03:25 GMT

Fluorescence Spectrometer Graph

←Older revision		Revision as of 07:03, 26 October 2012
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	\| 1\|\|30 μL\|\|900 μL\|\|176 μL\|\|1.8 mL		\| 1\|\|30 μL\|\|900 μL\|\|176 μL\|\|1.8 mL
	\|-		\|-
-
-

Mary Mendoza: /* Chemiluminescence of Luminol */

Mary Mendoza — Fri, 26 Oct 2012 05:06:42 GMT

Chemiluminescence of Luminol

←Older revision		Revision as of 05:06, 26 October 2012
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	* The lamp of LS 55 Perkin Elmer fluorescence spectrometer was turned off. The signal was set to a function of time. The wavelength was set at 425 nm. The excitation slit was placed at 15 nm while the emission slit was placed at 20 nm. The duration of data collection was settled at 300 sec.		* The lamp of LS 55 Perkin Elmer fluorescence spectrometer was turned off. The signal was set to a function of time. The wavelength was set at 425 nm. The excitation slit was placed at 15 nm while the emission slit was placed at 20 nm. The duration of data collection was settled at 300 sec.
		+	* The reagents were transferred into the cuvette using automatic delivery micropipets. The HRP was added last for this initiates the reaction through catalysis of hydrogen peroxide.
		+	* The collected data are shown in the next section.

	==Fluorescence Spectrometer Graph==		==Fluorescence Spectrometer Graph==

Mary Mendoza: /* Chemiluminescence of Luminol */

Mary Mendoza — Fri, 26 Oct 2012 05:02:33 GMT

Chemiluminescence of Luminol

←Older revision		Revision as of 05:02, 26 October 2012
Line 53:		Line 53:

	Image 2. Below each pH paper is shown the number of drops of 1 M HCl into the solution of luminol. The light green color corresponds to the pH level of 7 to 8.		Image 2. Below each pH paper is shown the number of drops of 1 M HCl into the solution of luminol. The light green color corresponds to the pH level of 7 to 8.
		+
		+
		+	* The lamp of LS 55 Perkin Elmer fluorescence spectrometer was turned off. The signal was set to a function of time. The wavelength was set at 425 nm. The excitation slit was placed at 15 nm while the emission slit was placed at 20 nm. The duration of data collection was settled at 300 sec.

	==Fluorescence Spectrometer Graph==		==Fluorescence Spectrometer Graph==

Mary Mendoza: /* Chemiluminescence of Luminol */

Mary Mendoza — Fri, 26 Oct 2012 04:56:02 GMT

Chemiluminescence of Luminol

←Older revision		Revision as of 04:56, 26 October 2012
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	[[Image:2012-10-09 14.59.55.jpg]]		[[Image:2012-10-09 14.59.55.jpg]]

-	Image 2. Below each pH paper is shown the number of drops of 1 M HCl into the solution of luminol.	+	Image 2. Below each pH paper is shown the number of drops of 1 M HCl into the solution of luminol. The light green color corresponds to the pH level of 7 to 8.

	==Fluorescence Spectrometer Graph==		==Fluorescence Spectrometer Graph==

Mary Mendoza: /* Chemiluminescence of Luminol */

Mary Mendoza — Fri, 26 Oct 2012 04:54:20 GMT

Chemiluminescence of Luminol

←Older revision		Revision as of 04:54, 26 October 2012
Line 48:		Line 48:

	Image 1. The navy blue color is attributed to a pH of 10 to 11.		Image 1. The navy blue color is attributed to a pH of 10 to 11.
		+
		+
		+	[[Image:2012-10-09 14.59.55.jpg]]
		+
		+	Image 2. Below each pH paper is shown the number of drops of 1 M HCl into the solution of luminol.

	==Fluorescence Spectrometer Graph==		==Fluorescence Spectrometer Graph==

Mary Mendoza: /* Chemiluminescence of Luminol */

Mary Mendoza — Fri, 26 Oct 2012 04:50:58 GMT

Chemiluminescence of Luminol

←Older revision		Revision as of 04:50, 26 October 2012
Line 46:		Line 46:

	[[Image:2012-10-09 14.17.45.jpg]]		[[Image:2012-10-09 14.17.45.jpg]]
		+
	Image 1. The navy blue color is attributed to a pH of 10 to 11.		Image 1. The navy blue color is attributed to a pH of 10 to 11.

Mary Mendoza: /* Chemiluminescence of Luminol */

Mary Mendoza — Fri, 26 Oct 2012 04:50:24 GMT

Chemiluminescence of Luminol

←Older revision		Revision as of 04:50, 26 October 2012
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	* Sodium bicarbonate precipitated in the solution. Addition of 4 mL of water diluted luminol from 30 mM to 10 mM.		* Sodium bicarbonate precipitated in the solution. Addition of 4 mL of water diluted luminol from 30 mM to 10 mM.
	* A pH paper was used to estimate the pH level of the solution since the electrode of the pH meter did not fit into the falcon tube of the solution.		* A pH paper was used to estimate the pH level of the solution since the electrode of the pH meter did not fit into the falcon tube of the solution.
-	*	+	* The images of the pH paper during qualitative analysis of the solution are embedded below:
		+
		+
		+	[[Image:2012-10-09 14.17.45.jpg]]
		+	Image 1. The navy blue color is attributed to a pH of 10 to 11.

	==Fluorescence Spectrometer Graph==		==Fluorescence Spectrometer Graph==

Mary Mendoza: /* Chemiluminescence of Luminol */

Mary Mendoza — Fri, 26 Oct 2012 04:46:55 GMT

Chemiluminescence of Luminol

←Older revision		Revision as of 04:46, 26 October 2012
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	* The reaction of 3-aminophthalhydrazide (luminol) and hydrogen peroxide is catalyzed by the addition of horseradish peroxidase (HRP) producing 3-aminophthalate.		* The reaction of 3-aminophthalhydrazide (luminol) and hydrogen peroxide is catalyzed by the addition of horseradish peroxidase (HRP) producing 3-aminophthalate.
	* The reagents originated from the HRP Assay of aminoantipyrine (AAP) laboratory period [[User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/10/02\|10/02/2012.]] For this procedure, 4-iodophenol, hydrogen peroxide, HRP, and water will be used. The AAP will be replaced in the reaction with luminol.		* The reagents originated from the HRP Assay of aminoantipyrine (AAP) laboratory period [[User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/10/02\|10/02/2012.]] For this procedure, 4-iodophenol, hydrogen peroxide, HRP, and water will be used. The AAP will be replaced in the reaction with luminol.
-	* ~~It was decided to prepare~~ 10 mM of luminol ~~since this~~ will be further diluted down to 3 mM upon the addition of water in the cuvette.	+	* A concentration of 10 mM of luminol was used. This will be further diluted down to 3 mM upon the addition of water in the cuvette.
	* The following calculations were made for luminol (FW 177.16):		* The following calculations were made for luminol (FW 177.16):

←Older revision		Revision as of 13:56, 7 December 2012
Line 67:		Line 67:

	{\| {{table}}		{\| {{table}}
		+	\| align="center" style="background:#f0f0f0;"\|''''
	\| align="center" style="background:#f0f0f0;"\|''HRP''		\| align="center" style="background:#f0f0f0;"\|''HRP''
	\| align="center" style="background:#f0f0f0;"\|''4-iodophenol''		\| align="center" style="background:#f0f0f0;"\|''4-iodophenol''
Line 73:		Line 74:
	\| align="center" style="background:#f0f0f0;"\|''H<sub>2</sub>O''		\| align="center" style="background:#f0f0f0;"\|''H<sub>2</sub>O''
	\|-		\|-
-	\| 30 μL\|\|54 μL\|\|900 μL\|\|176 μL\|\|1.8 mL	+	\| Molarity of Stock Solutions\|\|2.3 μM\|\|18 mM\|\|10 mM\|\|1.7 mM\|\|neutral
		+	\|-
		+	\| Volume\|\|30 μL\|\|54 μL\|\|900 μL\|\|176 μL\|\|1.8 mL
	\|-		\|-

←Older revision		Revision as of 07:06, 26 October 2012
Line 67:		Line 67:

	{\| {{table}}		{\| {{table}}
-	\| align="center" style="background:#f0f0f0;"\|''~~sample~~''	+	\| align="center" style="background:#f0f0f0;"\|''HRP''
	\| align="center" style="background:#f0f0f0;"\|''4-iodophenol''		\| align="center" style="background:#f0f0f0;"\|''4-iodophenol''
	\| align="center" style="background:#f0f0f0;"\|''luminol''		\| align="center" style="background:#f0f0f0;"\|''luminol''
Line 73:		Line 73:
	\| align="center" style="background:#f0f0f0;"\|''H<sub>2</sub>O''		\| align="center" style="background:#f0f0f0;"\|''H<sub>2</sub>O''
	\|-		\|-
-	\| 1\|\|30 μL\|\|900 μL\|\|176 μL\|\|1.8 mL	+	\| 30 μL\|\|54 μL\|\|900 μL\|\|176 μL\|\|1.8 mL
	\|-		\|-