Difference between revisions of "Team:UCSC/Safety"
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<style> | <style> | ||
| − | . | + | .metabolic-pathway { |
| − | margin | + | margin: auto; |
| − | + | ||
| − | + | ||
background: honeydew; | background: honeydew; | ||
| + | width: 100%; | ||
} | } | ||
h1 { | h1 { | ||
| − | font-family: 'objektiv-mk1' !important; | + | font-family: 'objektiv-mk1'!important; |
| − | font-size: 300%; | + | font-size: 300%; /*!important*/ |
font-weight: 300 !important; | font-weight: 300 !important; | ||
| + | width: 90%; | ||
} | } | ||
h2 { | h2 { | ||
font-family: 'objektiv-mk1' !important; | font-family: 'objektiv-mk1' !important; | ||
| − | font-size: | + | font-size: 200%; |
font-weight: 300; | font-weight: 300; | ||
| − | |||
| − | |||
} | } | ||
h3 { | h3 { | ||
font-family: 'objektiv-mk1' !important; | font-family: 'objektiv-mk1' !important; | ||
| − | font-size: | + | font-size: 120%; |
| − | font-weight: | + | font-weight: 200; |
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.proj-button { | .proj-button { | ||
position: relative; | position: relative; | ||
| − | width: | + | width: 25%; |
| + | text-decoration: none !important; | ||
} | } | ||
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opacity: 1; | opacity: 1; | ||
display: inline-block; | display: inline-block; | ||
| − | width: | + | width: 30%; |
| + | height: auto; | ||
| + | transition: .5s ease; | ||
| + | backface-visibility: hidden; | ||
| + | } | ||
| + | |||
| + | .proj-button-image-solo { | ||
| + | opacity: 1; | ||
| + | display: inline-block; | ||
| + | width: 65%; | ||
height: auto; | height: auto; | ||
transition: .5s ease; | transition: .5s ease; | ||
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transform: translate(-50%, -50%); | transform: translate(-50%, -50%); | ||
-ms-transform: translate(-50%, -50%) | -ms-transform: translate(-50%, -50%) | ||
| + | } | ||
| + | |||
| + | .overlap-button-text { | ||
| + | background-color: transparent; | ||
| + | color: black; | ||
| + | font-size: 20px; | ||
| + | font-family: "Objektiv-mk1" !important; | ||
| + | font-weight: 600 !important; | ||
| + | padding: 16px 32px; | ||
} | } | ||
.proj-button:hover .proj-button-image { | .proj-button:hover .proj-button-image { | ||
| + | opacity: 0.3; | ||
| + | } | ||
| + | |||
| + | .proj-button:hover .proj-button-image-solo { | ||
opacity: 0.3; | opacity: 0.3; | ||
} | } | ||
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} | } | ||
| − | . | + | .text { |
| − | + | background-color: transparent; | |
| − | + | color: black; | |
| + | font-size: 20px; | ||
| + | padding: 16px 32px; | ||
} | } | ||
| − | . | + | .paragraph-left { |
| − | font-family: 'objektiv-mk1'; | + | font-family: 'objektiv-mk1' !important; |
| − | font-size: | + | font-size: 18px; |
text-align: left; | text-align: left; | ||
| − | + | vertical-align: middle; | |
| − | + | ||
| − | + | ||
} | } | ||
p { | p { | ||
font-family: 'objektiv-mk1' !important; | font-family: 'objektiv-mk1' !important; | ||
| + | /* margin-left: 200px !important; | ||
| + | margin-right: 200px !important;*/ | ||
font-size: large !important; | font-size: large !important; | ||
} | } | ||
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.firstword { | .firstword { | ||
font-family: 'objektiv-mk1' !important; | font-family: 'objektiv-mk1' !important; | ||
| − | font-size: | + | font-size: 100px; |
| + | } | ||
| + | .quote { | ||
| + | font-family: 'objektiv-mk1' !important; | ||
| + | font-style: italic !important; | ||
| + | font-size: 28px; | ||
| + | line-height: 130%; | ||
| + | } | ||
| + | |||
| + | .quote-person { | ||
| + | font-family: 'objektiv-mk1' !important; | ||
| + | font-size: 22px; | ||
| + | width: 100% !important; | ||
| + | text-align: right !important; | ||
| + | padding-right: 80px !important; | ||
} | } | ||
.titleimg { | .titleimg { | ||
vertical-align: middle; | vertical-align: middle; | ||
| − | width: | + | width: 20%; |
} | } | ||
.titlebox { | .titlebox { | ||
| − | margin-left: | + | margin-left: 0px !important; |
} | } | ||
| − | . | + | .reference-list { |
| − | + | font-family: 'objektiv-mk1'; | |
| − | + | font-size: 12px; | |
| − | margin: | + | text-align: left; |
| + | list-style-type: none; | ||
| + | line-height: 100%; | ||
| + | margin-bottom: 80px; | ||
} | } | ||
| + | |||
| + | .reference-italic { | ||
| + | font-family: 'objektiv-mk1'; | ||
| + | font-size: 12px; | ||
| + | font-style: italic; | ||
| + | } | ||
| + | |||
| + | figcaption { | ||
| + | font-family: 'objektiv-mk1'; | ||
| + | font-style: italic; | ||
| + | font-size: 13px; | ||
| + | } | ||
| + | |||
@media (min-width: 1144px) { | @media (min-width: 1144px) { | ||
.container { | .container { | ||
| − | + | width: 100% !important; | |
| − | + | } | |
| + | |||
.text-container { | .text-container { | ||
| − | width: | + | width: 70%; |
padding-left: 15px; | padding-left: 15px; | ||
padding-right: 15px; | padding-right: 15px; | ||
} | } | ||
| + | |||
.titlebox { | .titlebox { | ||
width: 80%; | width: 80%; | ||
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@media (max-width: 1144px) { | @media (max-width: 1144px) { | ||
| − | + | .titlebox { | |
| − | margin-left: auto !important; | + | margin-left: auto !important; |
| + | } | ||
| + | |||
| + | p { | ||
| + | font-family: 'objektiv-mk1' !important; | ||
| + | margin-left: 10px !important; | ||
| + | margin-right: 10px !important; | ||
| + | font-size: medium !important; | ||
| + | } | ||
| + | |||
| + | .quote { | ||
| + | font-family: 'objektiv-mk1' !important; | ||
| + | font-style: italic !important; | ||
| + | font-size: 26px; | ||
| + | } | ||
| + | |||
| + | .quote-person { | ||
| + | font-family: 'objektiv-mk1' !important; | ||
| + | font-size: 20px; | ||
| + | width: 100%; | ||
| + | text-align: right; | ||
| + | padding-right: 80px !important; | ||
| + | } | ||
| + | |||
| + | .metabolic-pathway { | ||
| + | margin: auto; | ||
| + | background: honeydew; | ||
| + | width: 100%; | ||
| + | } | ||
| + | |||
| + | .container { | ||
| + | width: 80% !important; | ||
| + | padding-left: 0px !important; | ||
| + | padding-right: 0px !important; | ||
} | } | ||
.text-container { | .text-container { | ||
width: 80%; | width: 80%; | ||
| − | padding-left: | + | padding-left: 0px; |
| − | padding-right: | + | padding-right: 0px; |
} | } | ||
| − | + | figcaption { | |
| − | + | font-size: 14px; | |
| − | + | } | |
} | } | ||
| − | + | @media (max-width: 992px) { | |
| − | + | .proj-button-image-solo { | |
| − | + | opacity: 1; | |
| − | + | display: inline-block; | |
| − | + | width: 30%; | |
| − | + | height: auto; | |
| + | transition: .5s ease; | ||
| + | backface-visibility: hidden; | ||
| + | } | ||
} | } | ||
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opacity: 1; | opacity: 1; | ||
display: inline-block; | display: inline-block; | ||
| − | width: | + | width: 32%; |
| + | height: auto; | ||
| + | transition: .5s ease; | ||
| + | backface-visibility: hidden; | ||
| + | } | ||
| + | |||
| + | .proj-button-image-solo { | ||
| + | opacity: 1; | ||
| + | display: inline-block; | ||
| + | width: 35%; | ||
height: auto; | height: auto; | ||
transition: .5s ease; | transition: .5s ease; | ||
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transform: translate(-50%, -50%); | transform: translate(-50%, -50%); | ||
-ms-transform: translate(-50%, -50%) | -ms-transform: translate(-50%, -50%) | ||
| + | } | ||
| + | |||
| + | .overlap-button-text { | ||
| + | background-color: rgba(94, 94, 94, 0.4); | ||
| + | border-radius: 60px; | ||
| + | color: white; | ||
} | } | ||
h1 { | h1 { | ||
font-family: 'objektiv-mk1'; /*!important*/ | font-family: 'objektiv-mk1'; /*!important*/ | ||
| − | font-size: | + | font-size: 220%; /*!important*/ |
| + | width: 80%; | ||
} | } | ||
| − | + | h2 { | |
font-family: 'objektiv-mk1'; /*!important*/ | font-family: 'objektiv-mk1'; /*!important*/ | ||
| − | font-size: | + | font-size: 20px; /*!important*/ |
} | } | ||
| − | + | ||
| − | . | + | .quote { |
| − | + | font-family: 'objektiv-mk1' !important; | |
| + | font-style: italic !important; | ||
| + | font-size: 20px; | ||
} | } | ||
| − | . | + | .quote-person { |
width: 100%; | width: 100%; | ||
| − | + | font-family: 'objektiv-mk1' !important; | |
| − | + | font-size: 16px; | |
| + | text-align: right !important; | ||
| + | padding-right: 20px !important; | ||
} | } | ||
| − | + | .text-container { | |
| − | width: | + | width: 82%; |
| + | padding-left: 0px; | ||
| + | padding-right: 0px; | ||
} | } | ||
| − | + | figcaption { | |
| − | + | font-size: 14px; | |
| + | line-height: 140%; | ||
} | } | ||
| − | . | + | .reference-list { |
| − | + | font-family: 'objektiv-mk1'; | |
| − | + | font-size: 12px; | |
| − | + | text-align: left; | |
| − | + | list-style-type: none; | |
| − | + | line-height: 100%; | |
| − | + | margin-bottom: 80px; | |
| − | + | margin-right: -70px; | |
| − | + | margin-left: -70px; | |
} | } | ||
| + | } | ||
| + | /* p { | ||
| + | margin-left: 10px !important; | ||
| + | margin-right: 10px !important; | ||
| + | } | ||
| + | }*/ | ||
| − | + | @media (max-width: 576px) { | |
| − | + | ||
| − | + | ||
| − | + | ||
.proj-button-image { | .proj-button-image { | ||
opacity: 1; | opacity: 1; | ||
display: inline-block; | display: inline-block; | ||
| − | width: | + | width: 50%; |
| + | height: auto; | ||
| + | transition: .5s ease; | ||
| + | backface-visibility: hidden; | ||
| + | } | ||
| + | |||
| + | .proj-button-image-solo { | ||
| + | opacity: 1; | ||
| + | display: inline-block; | ||
| + | width: 50%; | ||
height: auto; | height: auto; | ||
transition: .5s ease; | transition: .5s ease; | ||
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transform: translate(-50%, -50%); | transform: translate(-50%, -50%); | ||
-ms-transform: translate(-50%, -50%) | -ms-transform: translate(-50%, -50%) | ||
| + | } | ||
| + | |||
| + | .overlap-button-text { | ||
| + | background-color: rgba(94, 94, 94, 0.5); | ||
| + | border-radius: 60px; | ||
| + | color: white; | ||
| + | font-size: 12px; | ||
} | } | ||
.firstword { | .firstword { | ||
font-family: 'objektiv-mk1' !important; | font-family: 'objektiv-mk1' !important; | ||
| − | font-size: | + | font-size: 70px; |
} | } | ||
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vertical-align: middle; | vertical-align: middle; | ||
width: 40%; | width: 40%; | ||
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} | } | ||
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} | } | ||
| − | + | h1 { | |
| − | + | font-size: 160%; | |
| − | + | ||
} | } | ||
| − | . | + | .quote { |
| − | margin-bottom: 15px; | + | font-family: 'objektiv-mk1' !important; |
| + | font-style: italic !important; | ||
| + | font-size: 17px; | ||
| + | } | ||
| + | |||
| + | .quote-person { | ||
| + | width: 60% !important; | ||
| + | font-family: 'objektiv-mk1' !important; | ||
| + | font-size: 12px; | ||
| + | text-align: right !important; | ||
| + | margin-right: -120px !important; | ||
| + | } | ||
| + | |||
| + | .paragraph-left { | ||
| + | font-size: 13px; | ||
| + | line-height: 170%; | ||
| + | } | ||
| + | |||
| + | .text-container { | ||
| + | width: 85%; | ||
| + | padding-left: 0px; | ||
| + | padding-right: 0px; | ||
| + | } | ||
| + | |||
| + | .metabolic-pathway { | ||
| + | margin: auto; | ||
| + | background: honeydew; | ||
| + | width: 110%; | ||
| + | margin-left: -20px; | ||
| + | margin-right: -20px; | ||
| + | } | ||
| + | |||
| + | figcaption { | ||
| + | font-size: 12px; | ||
| + | line-height: 130%; | ||
| + | } | ||
| + | |||
| + | .reference-list { | ||
| + | font-family: 'objektiv-mk1'; | ||
| + | font-size: 12px; | ||
| + | text-align: left; | ||
| + | list-style-type: none; | ||
| + | line-height: 100%; | ||
| + | margin-bottom: 80px; | ||
| + | margin-right: -50px; | ||
| + | margin-left: -50px; | ||
| + | } | ||
| + | } | ||
| + | |||
| + | @media (max-width: 400px) { | ||
| + | .quote { | ||
| + | font-size: 15px; | ||
} | } | ||
} | } | ||
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| − | <body> | + | <body style="background-color: honeydew;"> |
<div id="page"> | <div id="page"> | ||
<center> | <center> | ||
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<img class="titleimg" src="https://static.igem.org/mediawiki/2017/3/37/Gloves.png"> | <img class="titleimg" src="https://static.igem.org/mediawiki/2017/3/37/Gloves.png"> | ||
| − | <h1>SAFETY</h1><br><br> | + | <h1>SAFETY</h1> |
| + | |||
| + | <br> | ||
| + | <br> | ||
<div class="text-container"> | <div class="text-container"> | ||
| − | <p><span class="firstword">When</span> choosing an organism to help us produce acetaminophen and vitamin B | + | <p><span class="firstword">When</span> choosing an organism to help us produce acetaminophen and vitamin B<sub>12</sub> we sought out an FDA approved microbe; however, we resorted to using a non-toxic cyanobacteria as a model for spirulina. Spirulina is generally recognized as safe organism and our genes were chosen from bio-safety level 1 (BSL-1) organisms.</p> |
<div class="container"> | <div class="container"> | ||
<div class="row"> | <div class="row"> | ||
| − | <h2><b>L A B</b> SAFETY</h2><br> | + | <h2><b>L A B</b> SAFETY</h2> |
| + | <br> | ||
<div class="col-md-1"></div> | <div class="col-md-1"></div> | ||
<div class="col-md-5"> | <div class="col-md-5"> | ||
| Line 345: | Line 517: | ||
<br> | <br> | ||
| − | <h2><b>P R O J E C T</b> SAFETY</h2><br> | + | <h2><b>P R O J E C T</b> SAFETY</h2> |
| + | |||
| + | <br> | ||
| − | <p>We are growing two different strains of <i>S. elongatus</i>: 7002 and 7942. While PCC 7942 will act as our host organism due to its metabolic similarity to <i>A. platensis</i>, we plan to use PCC 7002, which requires external | + | <p>We are growing two different strains of <i>S. elongatus</i>: 7002 and 7942. While PCC 7942 will act as our host organism due to its metabolic similarity to <i>A. platensis</i>, we plan to use PCC 7002, which requires external B<sub>12</sub> for survival, as a growth reference for the detection of vitamin B<sub>12</sub>. PCC 7942 was obtained from the University of California, San Diego (UCSD) iGEM team and PCC 7002 was obtained from Dr. Donald Bryant at Pennsylvania State University.</p> |
| − | <h3>VITAMIN B | + | <br> |
| + | <br> | ||
| + | |||
| + | <h3>VITAMIN B<sub>12</sub></h3> | ||
| + | |||
| + | <br> | ||
| − | <h4><b>G E N E S</b> U S E D:<h4> | + | <h4><b>G E N E S</b> U S E D:</h4> |
| − | <p style=" | + | <p style="padding-left: 30px; padding-right: 30px;"><b>I. <i>ssuE</i></b> --> produces FMN reductase<br><br> |
| − | + | ||
| − | + | ||
| − | "><b>I. <i>ssuE</i></b> | + | |
| − | Organism: <i>Synechoccocous elongatus</i> strain PCC 7002<br> | + | Organism: <i>Synechoccocous elongatus</i> strain PCC 7002 |
| − | Biosafety <b>Level I</b><br><br> | + | <br> |
| + | Biosafety <b>Level I</b> | ||
| + | <br> | ||
| + | <br> | ||
| − | The synthesis of 5,6-DMB first requires the presence of reduced flavin mononucleotide ( | + | The synthesis of 5,6-DMB first requires the presence of reduced flavin mononucleotide (FMNH<sub>2</sub>). FMNH<sub>2</sub> is reduced from flavin mononucleotide (FMN) by FMN reductase, which is encoded with the <i>ssuE</i> gene [19,20]. |
| + | <br> | ||
| + | <br> | ||
| − | + | FMNH<sub>2</sub> + NADP<sup>+</sup> = FMN + NADPH | |
| + | <br> | ||
| + | <br> | ||
| − | <b>II. <i>bluB</i></b> => 5,6-DMB synthase<br><br> | + | <b>II. <i>bluB</i></b> => 5,6-DMB synthase |
| + | <br> | ||
| + | <br> | ||
| − | Organism: <i>Sinorhizobium meliloti</i> 1021<br> | + | Organism: <i>Sinorhizobium meliloti</i> 1021 |
| + | <br> | ||
Biosafety <b>Level I</b> | Biosafety <b>Level I</b> | ||
</p> | </p> | ||
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<h3>ACETAMINOPHEN</h3> | <h3>ACETAMINOPHEN</h3> | ||
| − | <h4><b>G E N E S</b> U S E D:<h4> | + | <h4><b>G E N E S</b> U S E D:</h4> |
| − | <p style=" | + | <p style="padding-left: 30px; padding-right: 30px;"><b>I. <i>4ABH</i></b> --> 4-aminophenol<br><br> |
| − | + | ||
| − | + | ||
| − | "><b>I. <i>4ABH</i></b> | + | |
| − | Organism: <i>Agaricus bisporus</i><br> | + | Organism: <i>Agaricus bisporus</i> |
| − | Biosafety <b>Level I</b><br><br> | + | <br> |
| + | Biosafety <b>Level I</b> | ||
| + | <br> | ||
| + | <br> | ||
| − | <b>II. <i>nhoA</i></b> | + | <b>II. <i>nhoA</i></b> --> converts 4-aminophenol into acetaminophen |
| + | <br> | ||
| + | <br> | ||
| − | Organism: <i>Escherichia coli</i><br> | + | Organism: <i>Escherichia coli</i> |
| − | Biosafety <b>Level I</b><br><br><br> | + | <br> |
| + | Biosafety <b>Level I</b> | ||
| + | <br> | ||
| + | <br> | ||
| + | <br> | ||
</p> | </p> | ||
| − | <h3><b>T O X I C I T Y</b> TESTS</h3 | + | <h3><b>T O X I C I T Y</b> TESTS</h3> |
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
<br> | <br> | ||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| + | <p>In order to judge whether our project would possibly be safe for consumption, we performed toxicity tests for the acetaminophen pathway in PCC 7942. Varying amounts of acetaminophen were added to liquid culture and the OD of each sample was observed. Liquid culture samples containing 1.32 mM, 6.62 mM, and 13.23 mM of acetaminophen were left to oxidize overnight and the color change was compared with liquid cultures samples containing similar concentrations of 4-aminophenol.</p> | ||
| + | <br> | ||
| + | <div class="container"> | ||
| + | <div class="reference-list"> | ||
| + | <hr> | ||
| + | <li>[19] R. Caspi, T. Altman, J. M. Dale, K. Dreher, C. A. Fulcher, F. Gilham, P. Kaipa, A. S. Karthikeyan, A. Kothari, M. Krummenacker, M. Latendresse, L. A. Mueller, S. Paley, L. Popescu, A. Pujar, A. G. Shearer, P. Zhang, and P. D. Karp, “The MetaCyc database of metabolic pathways and enzymes and the BioCyc collection of pathway/genome databases,” Nucleic Acids Research, vol. 38, pp. D473–D479, Jan. 2010.</li> | ||
| + | <li>[20] “ssuE - FMN reductase (NADPH) - Escherichia coli (strain K12) - ssuE gene & protein.”</li> | ||
| + | </div> | ||
| + | </div> | ||
</div> | </div> | ||
</html> | </html> | ||
| − | |||
{{UCSC-Footer}} | {{UCSC-Footer}} | ||
Revision as of 00:26, 29 October 2017
SAFETY
When choosing an organism to help us produce acetaminophen and vitamin B12 we sought out an FDA approved microbe; however, we resorted to using a non-toxic cyanobacteria as a model for spirulina. Spirulina is generally recognized as safe organism and our genes were chosen from bio-safety level 1 (BSL-1) organisms.
L A B SAFETY
Proper PPE for BSL-1 organisms are worn in our laboratory. Within the lab space, every person is expected to be wearing gloves, long pants, close-toed shoes, a lab coat, and protective goggles. All personnel also must be accompanied with one other person so that there is no one alone in lab.
P R O J E C T SAFETY
We are growing two different strains of S. elongatus: 7002 and 7942. While PCC 7942 will act as our host organism due to its metabolic similarity to A. platensis, we plan to use PCC 7002, which requires external B12 for survival, as a growth reference for the detection of vitamin B12. PCC 7942 was obtained from the University of California, San Diego (UCSD) iGEM team and PCC 7002 was obtained from Dr. Donald Bryant at Pennsylvania State University.
VITAMIN B12
G E N E S U S E D:
I. ssuE --> produces FMN reductase
Organism: Synechoccocous elongatus strain PCC 7002
Biosafety Level I
The synthesis of 5,6-DMB first requires the presence of reduced flavin mononucleotide (FMNH2). FMNH2 is reduced from flavin mononucleotide (FMN) by FMN reductase, which is encoded with the ssuE gene [19,20].
FMNH2 + NADP+ = FMN + NADPH
II. bluB => 5,6-DMB synthase
Organism: Sinorhizobium meliloti 1021
Biosafety Level I
ACETAMINOPHEN
G E N E S U S E D:
I. 4ABH --> 4-aminophenol
Organism: Agaricus bisporus
Biosafety Level I
II. nhoA --> converts 4-aminophenol into acetaminophen
Organism: Escherichia coli
Biosafety Level I
T O X I C I T Y TESTS
In order to judge whether our project would possibly be safe for consumption, we performed toxicity tests for the acetaminophen pathway in PCC 7942. Varying amounts of acetaminophen were added to liquid culture and the OD of each sample was observed. Liquid culture samples containing 1.32 mM, 6.62 mM, and 13.23 mM of acetaminophen were left to oxidize overnight and the color change was compared with liquid cultures samples containing similar concentrations of 4-aminophenol.
