User:Allison K. Alix/Notebook/CHEM-581/2013/02/06: Difference between revisions
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|style="background-color: #EEE"|[[Image:BDLlogo_notext_lr.png|128px]]<span style="font-size:22px;"> | |style="background-color: #EEE"|[[Image:BDLlogo_notext_lr.png|128px]]<span style="font-size:22px;"> Microgel Preparation</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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==Procedure== | ==Procedure== | ||
1) Prepare 20mM phosphate buffer (pH=7.4) | |||
::Dissolve 1.256 g disodium phosphate and 1.337 g monosodium phosphate in ~900mL water. Add sodium hydroxide to obtain pH of 7.4 (~100mL) | |||
2) Dissolve 0.5 g PVOH (MW= 89,000-98,000 g/mol) in 3mL phosphate buffer. Heat @ 100°C for 15 minutes with stirring. | |||
3) Add 2μL R6G in 2mL phosphate buffer to the PVOH/buffer mixture. Be sure to mix thoroughly. | |||
4) Add 100mL Mineral Oil (CVS, 99.9%) to a homogenizer and subsequently add the PVOH mixture. Homogenize for ~5minutes to form a w/o emulsion | |||
5) Pour final w/o emulsion into a beaker and place in a freezer at -20°C for 20 hours. Remove and allow thawing for 4 hours. Repeat this cycle for a total of three cycles. | |||
==Calculations== | |||
'''Preparation of 20mM phosphate buffer, pH = 7.4''' | '''Preparation of 20mM phosphate buffer, pH = 7.4''' | ||
[Base] = monosodium phosphate, 119.98 g/mol | [Base] = monosodium phosphate, 119.98 g/mol | ||
[Acid] = disodium phosphate, 141.96 g/mol | [Acid] = disodium phosphate, 141.96 g/mol | ||
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11.14mmol x 119.98g/mol x 1mol/1000mmol = 1.337g monosodium phosphate | 11.14mmol x 119.98g/mol x 1mol/1000mmol = 1.337g monosodium phosphate | ||
==References== | ==References== | ||
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[[Category:Course]] | [[Category:Course]] | ||
Latest revision as of 22:25, 26 September 2017
 Microgel Preparation
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Objective-Begin PVA microsphere preparation Procedure1) Prepare 20mM phosphate buffer (pH=7.4)
2) Dissolve 0.5 g PVOH (MW= 89,000-98,000 g/mol) in 3mL phosphate buffer. Heat @ 100°C for 15 minutes with stirring. 3) Add 2μL R6G in 2mL phosphate buffer to the PVOH/buffer mixture. Be sure to mix thoroughly. 4) Add 100mL Mineral Oil (CVS, 99.9%) to a homogenizer and subsequently add the PVOH mixture. Homogenize for ~5minutes to form a w/o emulsion 5) Pour final w/o emulsion into a beaker and place in a freezer at -20°C for 20 hours. Remove and allow thawing for 4 hours. Repeat this cycle for a total of three cycles. CalculationsPreparation of 20mM phosphate buffer, pH = 7.4 [Base] = monosodium phosphate, 119.98 g/mol [Acid] = disodium phosphate, 141.96 g/mol pH = pKa + log ([Base]/[Acid]) 7.4 = 7.3 + log ([Base]/[Acid]) ([Base]/[Acid]) = 1.2589 [Acid] + [Base] = 20mM [Base] = 20mM - [Acid] (20mM - [Acid])/[Acid] = 1.2589 [disodium phosphate] = 8.85mM [monosodium phosphate] = 11.14mM 8.85mmol x 141.96g/mol x 1mol/1000mmol = 1.256g disodium phosphate 11.14mmol x 119.98g/mol x 1mol/1000mmol = 1.337g monosodium phosphate Referenceshttp://www.sciencedirect.com/science/article/pii/S0168365998000893
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