User:Alicia Rasines Mazo/Notebook/CHEM-571 Experimental Biological Chemistry/2014/09/24: 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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==Tasks for September 24== | ==Tasks for September 24== | ||
To run HPLC with a GPC column | To run HPLC with a GPC (Gel Permeation Chromatography) column | ||
== | ===Preparation of solutions for HPLC chromatography=== | ||
To make a solution of 0.5M acetic acid and 0.3M Na<sub>2</sub>SO<sub>4</sub>: | To make a solution of 0.5M acetic acid and 0.3M Na<sub>2</sub>SO<sub>4</sub>: | ||
*(0.5mol HAc/L)(0.5L)(60.05g/mol)(1mL/1.05g)=14.3mL of acetic acid need to added to 500 mL volumetric flask | *(0.5mol HAc/L)(0.5L)(60.05g/mol)(1mL/1.05g)=14.3mL of acetic acid need to be added (under fume hood) to 500 mL volumetric flask | ||
*(0.3mol NaSO<sub>4</sub>/L)(0.5L)(142.04g/mol)=21.3060 g of NaSO<sub>4</sub> need to be added to volumetric flask | *(0.3mol NaSO<sub>4</sub>/L)(0.5L)(142.04g/mol)=21.3060 g of NaSO<sub>4</sub> need to be added to volumetric flask | ||
*Make up to 500 mL with HPLC water | *Make up to 500 mL mark with HPLC water | ||
To make dilutions of 1g/L of lsozyme: | To make dilutions of 1g/L of lsozyme: | ||
*'''0.5g/L''': (1g/L)(v<sub>1</sub>)=(0.5g/L)(10 mL) | *'''0.5g/L''': (1g/L)(v<sub>1</sub>)=(0.5g/L)(10 mL) | ||
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*'''0.1g/L''':(1g/L)(v<sub>1</sub>)=(0.1g(L)(10 mL) | *'''0.1g/L''':(1g/L)(v<sub>1</sub>)=(0.1g(L)(10 mL) | ||
*v<sub>1</sub>=1 mL of 1g/L lysozyme needs to be added | *v<sub>1</sub>=1 mL of 1g/L lysozyme needs to be added | ||
*Make up remainder of volume to 10 mL with distilled water | |||
===GPC=== | |||
*Prior to running HPLC, ran membrane filtration of solvent. | |||
*Filtered 0.5 M HAc 0.3 M NaSO<sub>4</sub> under a vacuum to ensure all particles were <0.45 microns to degas (get rid of CO<sub>2</sub> gas) | |||
* HPLC machine: Waters 1525 Binary HPLC pump, flow of A at 0.8 mL/min | |||
*For lysozyme and standard samples, syringe filters were used prior to injection | |||
Latest revision as of 00:21, 27 September 2017
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Tasks for September 24To run HPLC with a GPC (Gel Permeation Chromatography) column Preparation of solutions for HPLC chromatographyTo make a solution of 0.5M acetic acid and 0.3M Na2SO4:
To make dilutions of 1g/L of lsozyme:
GPC
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