<p class='jove_content'>We would like to acknowledge Mr. Ryohei Suganuma and Ms. Sakie Tachibana who helped perform the research. This study was supported in part by the JSPS KAKENHI Grant Numbers 15K20664, 24500848, and 21500405. This study was also funded in part by the MEXT/JST Tenure Track Promotion Program. A part of this study was based on the Japanese Unexamined Patent Application Publication No. 2011-169881 and the Japanese Patent No. 5708911.</p>

<p class="jove_content">NOTE: The device, including the method of analysis, is covered by Japanese Unexamined Patent Application Publication No. 2011-169881 and the Japanese Patent No. 5708911. This study, including the protocol of the experiment with human subjects, was approved by the Medical Ethics Committee of Kanazawa University (#553-1).</p>
<p class="jove_title">1. Prerequisites for the Perspiration Monitoring Device</p>
<p class="jove_content">NOTE: Perform these steps only once before the first use.</p>
<ol>
	<li>Install the universal serial bus (USB)-serial port conversion interface drivers in the computer<sup>27</sup>. If the drivers are already installed, skip this step.</li>
	<li>Connect the USB-serial port conversion interface to the computer via USB cable. Wait for the automatic driver installation if any.</li>
	<li>Set the parameters of the conversion interface and check the ID number of the serial port as follows.
	<ol>
		<li>Open the device manager, and select [View] &#8594; [Devices by Type].</li>
		<li>Locate the &#34;Ports (COM &#38; LPT)&#34; heading, and expand the section to find the heading containing &#34;CP210x USB to UART Bridge&#34; in the list.</li>
		<li>Memorize the ID number of the port (<em>e.g.,</em> &#34;COM5&#34;) in the &#34;CP210x USB to UART Bridge&#34; heading.</li>
		<li>Double click the &#34;CP210x USB to UART Bridge,&#34; and open the &#34;Port Settings&#34; tab.</li>
		<li>Set the parameters as follows: &#34;Bits per second&#34; = 9600, &#34;Data bits&#34; = 8, &#34;Parity&#34; = &#34;None,&#34; &#34;Stop bits&#34; = 1, and &#34;Flow control&#34; = &#34;None.&#34;</li>
		<li>Click the &#34;OK&#34; button to close the window, and close the device manager.</li>
	</ol>
	</li>
</ol>
<p class="jove_title">2. Setup of the Perspiration Monitoring Device</p>
<p class="jove_content">NOTE: Set the recording settings as follows before using the device. Repeat these steps for each device if multiple devices are to be used.</p>
<ol>
	<li>Make sure the battery is not inserted in the perspiration monitoring device.</li>
	<li>Plug the white connector of the USB-serial port conversion interface to the receptacle at the top of the perspiration monitoring device.</li>
	<li>Execute the perspiration recording software, followed by setting the device parameters as follows.
	<ol>
		<li>Open the &#34;Settings&#34; tab.</li>
		<li>Set the &#34;COM&#34; number to the ID number of the serial port (see step 1.3.3).</li>
		<li>Click the &#34;Confirm Connection&#34; button. Check if the message &#34;Connected&#8230;&#34; appears.</li>
		<li>At the &#34;Data File Folder&#34; settings, choose the drive and select the folder where the perspiration data will be saved. To select the subfolder, double click the folder to open.</li>
		<li>Set the &#34;Number of holes&#34; to &#34;4.&#34;</li>
	</ol>
	</li>
	<li>Open the &#34;Measure &#38; Rec&#34; tab, and set the &#34;Time Interval&#34; to the desired sample time.</li>
	<li>Insert a battery to the perspiration monitoring device.</li>
	<li>Fill the device with dry silica gel (color should be blue or green if the color indicator is available), and close the lid. If the lid does not close completely, reduce the amount of silica gel.</li>
	<li>Click the &#34;Start Logging Now&#34; button in the perspiration recording software.<br />
	NOTE: Because the perspiration recording starts just after the disconnection, the perspiration monitoring device may be left connected until the monitoring is to be started.</li>
</ol>
<p class="jove_title">3. Setup for the Measurement of Sympathetic Skin Response (SSR)</p>
<p class="jove_content">NOTE: These steps are intended to monitor the sympathetic activities as the palmar SSR, and are not necessarily required for perspiration monitoring itself. SSR is a change of skin potential according to the sympathetic arousal stimulation such as upset and concentration<sup>28,29</sup>.</p>
<ol>
	<li>Cleanse the skin where the SSR is to be recorded with an alcohol swab.</li>
	<li>Put the anode, cathode, and grounding electrode on the palm, back of the hand, and wrist, respectively, by means of an electrode paste. Fix the electrodes with medical tape.</li>
	<li>Set the conditions of the instrumentation amplifier by turning the corresponding knobs of the amplifier as follows: Sensitivity = 1 mV/V, High cut filter (&#34;HI CUT&#34;) = 3 kHz, and Low cut filter (&#34;LO CUT&#34;) = 0.5 Hz.</li>
	<li>Execute the SSR recording software, and start the recording at a sampling rate of 200 Hz by clicking the &#34;Start Measurement&#34; button in the software.</li>
</ol>
<p class="jove_title">4. Recording of the Perspiration</p>
<ol>
	<li>Put a medical double-sided tape on the bottom of the perspiration monitoring device. When putting a tape, make sure the measurement windows (<em>i.e.</em>&#160;four holes at the bottom of the device) are not obstructed.</li>
	<li>Disconnect the perspiration monitoring device from the USB-serial port conversion interface. Immediately after the disconnection, observe the perspiration monitoring begin automatically. Make sure the LED lamp is blinking.</li>
	<li>Remove the release liner of the medical double-sided tape, and put the perspiration monitoring device on the skin where perspiration is to be monitored.</li>
	<li>Wait 10 min for stabilization of water vapor diffusion.<br />
	NOTE: Although this process cannot be monitored, the previous study has confirmed that waiting 10 min is sufficient for stable monitoring<sup>26</sup>.</li>
	<li>Start the examination (<em>e.g.</em>, sympathetic activity test).</li>
	<li>After the examination, remove the perspiration monitoring device from the skin. To stop the SSR recording, click the &#34;Stop Measurement&#34; button in the SSR recording software, and remove all electrodes from the skin.</li>
</ol>
<p class="jove_title">5. Perspiration Analysis</p>
<ol>
	<li>Connect the perspiration monitoring device to the USB-serial port conversion interface (as in step 2.2).</li>
	<li>Execute the perspiration monitoring software.</li>
	<li>Open the &#34;Settings&#34; tab, and click the &#34;Confirm Connection&#34; button.</li>
	<li>Open the &#34;Measure &#38; Rec&#34; tab, and click the &#34;Download&#34; button to save the raw data of the device in the specified folder (see step 2.3.4).</li>
	<li>Perform perspiration analysis as follows.
	<ol>
		<li>Open the &#34;Analysis&#34; tab, and click the &#34;Read Data File.&#34; Observe the &#34;Open File&#34; dialog pop up. Select the saved file in the step 5.4, and then click the &#34;Open&#34; button.<br />
		NOTE: The software automatically performs perspiration analysis based on the published method<sup>26</sup>, and the results will appear on the screen. At the same time, the perspiration data as the comma-separated values format will be saved in the same folder.</li>
	</ol>
	</li>
</ol>

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<p class='jove_content'>The author Fukuoka Masakazu is the president of the Rousette Strategy Inc., where the developed device was assembled. No financial support was received from either Fukuoka Masakazu or the Rousette Strategy Inc.</p>

<p class="jove_content">The aim of this article is to introduce the use of a novel, wireless perspiration monitoring device. Owing to the recent progress of engineering, more accurate, easy-to-handle methods for temporal perspiration monitoring have been proposed; the ventilated chamber method<sup>24,25</sup> and the vapor pressure diffusion method<sup>23</sup> are representative examples. However, the ventilated chamber method requires the use of dry nitrogen or a pump with desiccant to produce a dry atmosphere, and thus, the exterior tends to be large. The vapor pressure diffusion method can be adopted in a small exterior, although the open chamber system can be largely influenced by the outer atmosphere, and the closed chamber system has a problem with either water vapor saturation (the chamber is filled with a saturated water vapor) or considerable power consumption of a water vapor condenser (<em>e.g.</em>, Peltier device).</p>
<p class="jove_content">To address the situation, we have recently developed a novel wireless device for perspiration monitoring<sup>26</sup>. In the device, the water vapor from the skin can be captured by a desiccant, enabling a constant but natural flow. In the course of the water vapor traveling, the T/RH sensor captures the flow of the water vapor as a change in the temperature and relative humidity. By calculating such changes under Fick&#39;s law, we could estimate the temporal changes of perspiration amount (mg/cm<sup>2</sup>/min) with &#60;5% error, relative to the conventional ventilated chamber method<sup>26</sup>. The developed device can be used, for example, to monitor mental stress (<strong>Figure 2</strong>), thermal sweating, and ultimately the perspiration-related dysregulation in an easier manner.</p>
<p class="jove_content">The developed device is easy to handle, and so there are few points to consider. However, the perspiration measurement would fail if the silica gel is not dry. Therefore, before the experiment,&#160;the examiner should make sure that the silica gel is completely dry (<em>i.e.</em>&#160;the color is blue). A previous study has estimated the maximum duration of measurement being longer than 4 hr without changing the silica gel<sup>26</sup>. To re-nature silica gel, dry it in a drying oven until the color turns blue. A conventional microwave oven is also useful. The perspiration measurement would also fail if the attachment of the device is not sufficient. We recommend the usage of a &#34;doughnut-shaped&#34; double-sided tape.</p>
<p class="jove_content">Nonetheless, the developed device has a limitation. Because of the delicate calculation and the limit of desiccant absorptivity, the error of the amount of perspiration should be considered, especially at the higher level of perspiration. Although we have confirmed the error rate as being &#60;5% relative to the conventional method<sup>26</sup>, the absolute value of the calculated perspiration value should be handled with caution.</p>
<p class="jove_content">Owing to the small, simple, and wireless design of the device, the multipoint perspiration monitoring under unrestricted conditions can be possible. As shown in <strong>Figure 3</strong>, the different temporal profiles of perspiration can be detected on the different positions of the skin (<em>i.e.</em>&#160;palm and anterior chest) in daily life conditions (<em>e.g.</em>, talking, eating foods, driving a car, shopping, <em>etc</em>.). The device, therefore, might be utilized for the simultaneous detection of abnormal sweating derived both from the mental and thermal regulatory system malfunction. For example, the dysregulation of the peripheral nervous system in diabetes might be detected if the unbalanced sweating between the left and right soles were to be detected. We are now planning an observational study of perspiration profiles in hospitalized patients as a clinical experiment.</p>

<p class="jove_content">Human perspiration, generally known as &#34;sweating,&#34; is not just a mechanism for thermoregulation<sup>1</sup>, but it is also related to certain kinds of diseases. The etiology of abnormal perspiration is broad, including: heatstroke, hyper- or hypothyroidism<sup>2</sup>, brain infarction<sup>3</sup>, diabetes mellitus<sup>4</sup>, dysautonomia<sup>5</sup>, menopause (known as &#34;hot flash&#34;)<sup>6</sup>, cystic fibrosis<sup>7</sup>, Parkinson&#39;s disease<sup>8</sup>, and social anxiety disorder<sup>9</sup>. In light of the number of perspiration-related diseases, it has been considered beneficial to monitor perspiration rates for the early diagnosis or prediction of such diseases (<em>e.g.,</em> prevention of heatstroke) in a ubiquitous manner<sup>10</sup>.</p>
<p class="jove_content">To date, only a small number of devices for perspiration monitoring have been proposed. In early days, skin conductance and relative humidity were used for indirect indices of the amount of perspiration<sup>11,12</sup>. Most recently, several kinds of flexible, wearable sensors for perspiration monitoring have been proposed<sup>13-19</sup>, although they are intended for the analysis of sweat electrolytes rather than the amount or temporal pattern of perspiration. The calculation of water vapor diffusion has been utilized for a more quantitative method of monitoring water exchange from the skin<sup>20-23</sup>. However, this requires (1) the assumption that the outer atmosphere is still and constant<sup>20</sup>, (2) enough sensitivity to detect the natural flow of water vapor<sup>21,22</sup>, or (3) a coolant (<em>e.g.</em>, Peltier device that consumes a substantial amount of electricity) to condense water vapor to liquid<sup>23</sup>; thus, they might be difficult for daily and long-term monitoring. As an alternative, a ventilated chamber method was developed<sup>20,24,25</sup>. In the ventilated chamber method, dry nitrogen or dehumidified air is infiltrated in a small chamber adjacent to the skin from a nitrogen gas tank or a pump, and gas in the water vapor evaporated from the skin is collected. The amount of water vapor from the skin can be calculated from the difference of the humidity in the outlet and inlet gases. Although this method can estimate the amount of perspiration very precisely, a nitrogen gas tank or a mechanical pump is generally large enough to impede daily monitoring.</p>
<p class="jove_content">To address these drawbacks, we have recently developed a novel device for perspiration monitoring, in which a closed chamber with a desiccant-driven enforced water vapor flow, enabled sensitive and long-term monitoring<sup>26</sup>. This device consists of a cylindrical plastic exterior, temperature/relative humidity (T/RH) sensor with recording microprocessor, and silica gel (<strong>Figure 1</strong>). In principle, the outer atmosphere should not interfere with the water vapor flow, and a coolant or ventilating chamber is not required. Perspiration profiles can be obtained by solving equations using a spreadsheet software<sup>26</sup>. A previous study has only shown the principle of the developed device and has omitted the detailed method for how to use the device because of space limitations.</p>
<p class="jove_content">The objective of this article, therefore, is to show a detailed method of how to use the developed device for perspiration monitoring, by showing the recording of stress-induced palmar perspiration during the sympathetic activity test as an example.</p>

<table><tbody><tr><td>Required for perspiration monitoring</td><td></td><td></td><td></td></tr><tr><td>Perspiration monitoring device</td><td>Rousette Strategy Inc.</td><td>SNT-200</td><td></td></tr><tr><td>USB-serial port conversion interface</td><td>Rousette Strategy Inc.</td><td>UUI-200</td><td></td></tr><tr><td>Perspiration recording software</td><td>Rousette Strategy Inc.</td><td>TED99</td><td></td></tr><tr><td>Silica gel</td><td>Wako Pure Chemical Industries Ltd.</td><td>194-16665</td><td>Type A silica gel should be used.</td></tr><tr><td>Medical double-sided tape</td><td>3M</td><td>2181</td><td>Medical grade is recommended because of the attachment to the skin.</td></tr><tr><td>Computer</td><td></td><td></td><td>Requires Windows operating system.</td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Required for the monitoring of sympathetic skin response&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Instrumentation amplifier</td><td>Nihon Kohden Corp.</td><td>AB-611J</td><td></td></tr><tr><td>Amplifier chassis</td><td>Nihon Kohden Corp.</td><td>MEG-6108</td><td></td></tr><tr><td>Input box</td><td>Nihon Kohden Corp.</td><td>JB-610B</td><td></td></tr><tr><td>Alcohol swab</td><td>Suzuran Sanitary Goods Co., Ltd.</td><td>4545766050846</td><td></td></tr><tr><td>Electrodes</td><td>Nihon Kohden Corp.</td><td>NE-114A</td><td></td></tr><tr><td>Electrode paste</td><td>Nihon Kohden Corp.</td><td>Z-401CE</td><td></td></tr><tr><td>Medical tape</td><td>Nichiban Co., Ltd.</td><td>SG257</td><td></td></tr><tr><td>Analog signal interface</td><td>Micro Science K.K.</td><td>C BOX-014</td><td></td></tr><tr><td>Analog-to-digital converter</td><td>Micro Science K.K.</td><td>ADM-686PCI</td><td></td></tr><tr><td>SSR recording software</td><td>Matsuyama Advance Co., Ltd.</td><td>LaBDAQ2000</td><td></td></tr></tbody></table>

a detailed protocol for perspiration monitoring using a novel, small, wireless device

<p class="jove_content">Perspiration monitoring can be utilized for the detection of certain diseases, such as thermoregulation and mental disorders, particularly when the patients are unaware of such disorders or are having difficulty expressing their symptoms. Until now, several devices for perspiration monitoring have been developed; however, such devices tend to have a relatively large exterior, considerable power consumption, and/or less sensitivity.</p>
<p class="jove_content">Recently, we developed a small, wireless device for perspiration monitoring. The device consists of a temperature/relative humidity (T/RH) sensor, battery-driven small data logger, and silica gel as a desiccant in a small cylindrical exterior. The T/RH sensor is placed between the detection windows (through which the water vapor from the skin enters) and the silica gel. The underlying principle of the perspiration monitoring device is based on Fick&#39;s law of diffusion, which means that water vapor flux from the skin to the silica gel (<em>i.e.</em>&#160;transepidermal water loss and perspiration) can be captured by change in humidity at the T/RH sensor. In addition, a baseline subtraction method was adopted to distinguish perspiration and transepidermal water loss.</p>
<p class="jove_content">As shown in the previous report, the developed device can monitor the perspiration at any sites of the body in an easy, wireless manner. However, detailed methods of how to use the device have not been disclosed yet. In this article, therefore, we would like to show the point-by-point tutorials of how to use the device for perspiration monitoring, by showing the sympathetic activity test with the sympathetic skin response monitoring as an example.</p>

<p class="jove_content" fo:keep-together.within-page="1">Using this novel device for perspiration monitoring (<strong>Figure 1</strong>) and the Fick&#39;s law-based calculation, the temporal perspiration profiles can be obtained in an easy, wireless manner. <strong>Figure 2</strong> shows representative data of wireless perspiration monitoring during the sympathetic activity test. In the experiment, the device for the perspiration monitoring, along with the electrodes for the sympathetic skin response (SSR) monitoring, was attached to the subject&#39;s palm. For the sympathetic activity test, the subject was requested to sit and perform the following tasks: (1) take a deep inspiration 5 times with 1 min intervals, and (2) do a mental calculation (<em>e.g.</em>, continuously subtract 7 from 100, or the &#34;Flash Anzan&#34; in which the participant sums up the number displayed one after the other on a computer screen) to evoke sympathetic activity. The perspiration and the SSR were simultaneously recorded during the stress conditions. As a result of deep inspiration and mental calculation, the sympathetic activity-induced palmar perspiration could be measured using the developed device. <strong>Figure 3</strong> shows representative data of multipoint measurement during daily activities. About 1 hr recording of perspiration at the palm (emotional sweating) and the anterior chest (thermal sweating) showed distinct patterns according to the activities.</p>
<p class="jove_content" fo:keep-together.within-page="1"><img alt="Figure 1" src="/files/ftp_upload/54837/54837fig1.jpg" /><br />
<strong>Figure 1:</strong> <strong>The Novel Device for Perspiration Monitoring. </strong>(<strong>A</strong>) The exterior of the device containing dry silica gel. (<strong>B</strong>, <strong>C</strong>) A &#34;doughnut-shaped&#34; double-sided tape used in this study and its attachment to the device. (<strong>D</strong>) Attachment of the device to the skin. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54837/54837fig1large.jpg" target="_blank">Please click here to view a larger version of this figure.</a></p>
<p class="jove_content" fo:keep-together.within-page="1"><img alt="Figure 2" src="/files/ftp_upload/54837/54837fig2.jpg" /><br />
<strong>Figure 2:</strong> <strong>Representative Recording of Perspiration Under the Sympathetic Activity Test.</strong> (<strong>A</strong>) Attachment of the perspiration monitoring device with the sympathetic skin response (SSR) electrodes on the palm. (<strong>B</strong>) As a result of sympathetic activity test, palmar perspiration along with the SSR reaction was observed in response to the sympathetic activities. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54837/54837fig2large.jpg" target="_blank">Please click here to view a larger version of this figure.</a></p>
<p class="jove_content" fo:keep-together.within-page="1"><img alt="Figure 3" src="/files/ftp_upload/54837/54837fig3.jpg" /><br />
<strong>Figure 3:</strong> <strong>Representative Multipoint Recording of Perspiration During the Daily Activities.</strong> The distinct patterns of perspiration according to the different parts of the body (red line, at the palm; blue line, at the anterior chest) were observed during several activities (<em>e.g.</em>, going downstairs, driving a car, having a lunch with talking, and shopping). <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54837/54837fig3large.jpg" target="_blank">Please click here to view a larger version of this figure.</a></p>

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A Detailed Protocol for Perspiration Monitoring Using a Novel, Small, Wireless Device

<p class='jove_content'>Recently, we developed a small wireless device for perspiration monitoring. In this article, we present detailed protocols on how to use the device for perspiration monitoring with an example of the sympathetic activity test.</p>

Perspiration monitoring can be utilized for the detection of certain diseases, such as thermoregulation and mental disorders, particularly when the ...

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7.8K Views.  Kanazawa University. Recently, we developed a small wireless device for perspiration monitoring. In this article, we present detailed protocols on how to use the device for perspiration monitoring with an example of the sympathetic activity test.

Video: A Detailed Protocol for Perspiration Monitoring Using a Novel, Small, Wireless Device

Thin-Layer Chromatography

<p>Source: Lara Al Hariri and&#160;Ahmed Basabrain at the University of Massachusetts Amherst, MA, USA</p>
<ol class="note-items">
	<li data-note-item="1" data-parent-collapse="">Effects of Mobile Phase Composition on Compound Separation<button class="expand">Expand</button>
	<p>In this section of the lab, you will use TLC to compare how well pure hexane and a mixture of 60% hexane and 40% ethyl acetate can separate phenol, benzoic acid, and butyl phenyl ether by measuring the distance the compound traveled in each solvent.</p>
	<ul class="lab-items">
		<li data-sub-note-item="1-2">
		<div class="lab-step">Before starting the lab, put on a lab coat, safety glasses, and nitrile gloves. Note: Hexane and ethyl acetate are both volatile, toxic, and flammable, so always work with them in a fume hood. Change your gloves if you get solvent on them, as hexane and ethyl acetate will permeate nitrile gloves within minutes.</div>
		</li>
		<li data-sub-note-item="1-3">
		<div class="lab-step">Now, obtain a paper towel and lay it on the benchtop. Carefully pick up a TLC plate using tweezers at the very end of the plate.<strong> Note: </strong>Always hold TLC plates by the edges or at the very top with tweezers to avoid damaging or contaminating the silica gel.</div>
		</li>
		<li data-sub-note-item="1-4">
		<div class="lab-step">Look for chips or scratches in the silica gel and get a new plate if necessary. It&#39;s fine if there is a little chipping at only one end, since that can be the top.</div>
		</li>
		<li data-sub-note-item="1-5">
		<div class="lab-step">Place the plate on the paper towel in your fume hood, and lay a metric ruler next to it. Use a graphite pencil to gently make a small mark on the silica gel ~1 cm from the top of the plate. Write &#8216;100&#8217; on the plate above the mark.</div>
		</li>
		<li data-sub-note-item="1-6">
		<div class="lab-step">Next, gently mark the plate 1 cm above the bottom on both sides. Carefully trace a straight line between the bottom marks to make the starting line.</div>
		</li>
		<li data-sub-note-item="1-7">
		<div class="lab-step">Then, add 3 evenly spaced small marks to the starting line with the outer marks at least 0.5 cm from the sides of the plate. <strong>Note:</strong> It&#39;s important for the silica gel to be undamaged here, so if you scratch it with the pencil, prepare a new plate.</div>
		</li>
		<li data-sub-note-item="1-8">
		<div class="lab-step">Now, trace the outline of the TLC plate in your lab notebook and precisely copy the marks on the plate. Label the three marks as &#8216;phenol&#8217;, &#8216;benzoic acid&#8217;, and &#8216;butyl phenyl ether&#8217;. You may also want to label the marks on the plate itself.</div>
		</li>
		<li data-sub-note-item="1-9">
		<div class="lab-step">Prepare a second TLC plate labeled &#8216;60/40&#8217; in the same way. Now, bring the labeled plates to the common bench and locate the 1 mg/mL solutions of phenol, benzoic acid, and butyl phenyl ether in ethanol.</div>
		</li>
		<li data-sub-note-item="1-10">
		<div class="lab-step">Dip a clean capillary into one of the three solutions. Briefly and lightly dab the capillary squarely against the intersection between the starting line and the mark for that compound. You should spot twice for each compound.</div>
		</li>
		<li data-sub-note-item="1-11">
		<div class="lab-step">Use the same capillary to spot the same solution on your second TLC plate. The spots should be no more than 2 mm in diameter. Then, discard the capillary and close the vial.</div>
		</li>
		<li data-sub-note-item="1-12">
		<div class="lab-step">Spot the other two solutions on the plates, in the same way, using clean capillaries. Be careful not to cross-contaminate the solutions. <strong>Note</strong>: If a spot is very light or very small, wait for it to dry, and then spot the same solution on top of the dry spot. If the spot is still wet, capillary action could make it too wide and dilute.</div>
		</li>
		<li data-sub-note-item="1-13">
		<div class="lab-step">Once you have made three spots on each plate, discard the capillaries and bring the plates back to your hood. Keep the plates lying flat while they dry.</div>
		</li>
		<li data-sub-note-item="1-14">
		<div class="lab-step">Now, label a 50-mL jar &#8216;100% hexane&#8217; and bring the jar, its cap, and a 10-mL graduated cylinder to the solvent hood.</div>
		</li>
		<li data-sub-note-item="1-15">
		<div class="lab-step">Measure 2 &#8211; 3 mL of pure hexane with your graduated cylinder and pour it into the jar to fill it to a depth of 0.62 &#8211; 0.7 cm. Cap the bottle and jar and return to your fume hood.</div>
		</li>
		<li data-sub-note-item="1-16">
		<div class="lab-step">Label a second 50-mL jar &#8216;60% hexane, 40% ethyl acetate&#8217; and bring it and a clean graduated cylinder to the solvent hood. Measure 2 &#8211; 3 mL of the 60/40 mixture into the jar and return to your fume hood.</div>
		</li>
		<li data-sub-note-item="1-17">
		<div class="lab-step">Stand the plates next to the jars to compare the relative position of the solvent and the spots. The solvent level should be below the spots for each plate.</div>
		</li>
		<li data-sub-note-item="1-18">
		<div class="lab-step">Then, obtain three pieces of filter paper, put a filter paper upright against the wall in each jar, and then cap the jars tightly.</div>
		</li>
		<li data-sub-note-item="1-19">
		<div class="lab-step">Initial each plate above the mark so you can identify them later. Confirm that the filter papers in the jars have become saturated with solvent and that the spots on the plates are completely dry.</div>
		</li>
		<li data-sub-note-item="1-20">
		<div class="lab-step">Now, open the 100% hexane jar and use tweezers to gently place the 100 plate in the jar, keeping the plate level with the solvent surface as it enters the solvent. Close the jar tightly, being careful not to jostle the plate.</div>
		</li>
		<li data-sub-note-item="1-21">
		<div class="lab-step">Leave the jar still as the solvent travels up the plate. Development in pure hexane usually takes about 7 &#8211; 10 min. Keep an eye on the plate to make sure that the solvent doesn&#39;t reach the top.</div>
		</li>
		<li data-sub-note-item="1-22">
		<div class="lab-step">When the pure hexane solvent front is about 1 cm from the top of the plate, retrieve it using tweezers. Lay it on the paper towel with the silica gel facing up and quickly mark the solvent front with a line and pencil across the plate. Leave the plate lying flat as the solvent evaporates.</div>
		</li>
		<li data-sub-note-item="1-23">
		<div class="lab-step">Place the other plate in the 60/40 eluent in the same way. Wait until the solvent front of the 60/40 plate is about 1 cm from the top, and then retrieve it and quickly mark the solvent front in the same way.</div>
		</li>
		<li data-sub-note-item="1-24">
		<div class="lab-step">Once the 100% hexane plate is completely dry, pick it up with tweezers and bring it and a pencil to the UV lamp on the common bench.</div>
		</li>
		<li data-sub-note-item="1-25">
		<div class="lab-step">Place the plate under the lamp and illuminate it with UV light. The plate will glow green because of a fluorescent marker in the silica gel layer. The dark spots are the compounds.</div>
		</li>
		<li data-sub-note-item="1-26">
		<div class="lab-step">Carefully outline every spot in pencil, even if they haven&#39;t moved very far.</div>
		</li>
		<li data-sub-note-item="1-27">
		<div class="lab-step">Then, turn off the lamp and bring your plate to the iodine development hood. Open the iodine chamber. Use tweezers to gently place the plate upright in the iodine chamber, and then cover the chamber again.</div>
		</li>
		<li data-sub-note-item="1-28">
		<div class="lab-step">Wait 3 &#8211; 4 min for the iodine vapor to stay in the spots. Then, retrieve the plate with tweezers, close the chamber, and outline the brown and yellow spots on the plate. These spots will fade, so mark them quickly.</div>
		</li>
		<li data-sub-note-item="1-29">
		<div class="lab-step">Visualize the dry 60/40 plate under UV light and then with iodine vapor in the same way.<strong> Note: </strong>To avoid side reactions, always check plates under UV light before exposing them to iodine.</div>
		</li>
		<li data-sub-note-item="1-30">
		<div class="lab-step">Now, precisely copy the spots and the solvent fronts onto the corresponding drawings of the plates in your lab notebook. For each plate, measure and record the height of the solvent front relative to the starting line.</div>
		</li>
		<li data-sub-note-item="1-31">
		<div class="lab-step">Lastly, measure the distance between the center of each spot and its starting line, and record it in your lab notebook. You can now move on to the next part of the lab.</div>
		</li>
	</ul>
	</li>
	<li data-note-item="2" data-parent-collapse="">Unknown Compound Identification<button class="expand">Expand</button>
	<p>In the second half of the lab, you&#39;ll use TLC to find out what&#39;s in an unknown analgesic. It could contain one or more of these compounds: aspirin, caffeine, and acetaminophen. These compounds are highly polar, so you&#39;ll use a premade mixture of 90% ethyl acetate and 10% hexane as the eluent.</p>
	<p>You&#39;ll run the three reference compounds and the unknown all on the same plate. Later, you&#39;ll use the Rf values of the reference compounds to identify the unknown. First, double-check which unknown material you were assigned to analyze before starting.</p>
	<ul class="lab-items">
		<li data-sub-note-item="2-3">
		<div class="lab-step">Now, lay a clean paper towel in the hood and obtain a new TLC plate. Prepare the top mark and the starting line the same way as before. Make four evenly spaced ticks on the plate, making sure that the outer marks are at least 0.5 cm away from the edges.</div>
		</li>
		<li data-sub-note-item="2-4">
		<div class="lab-step">Draw an exact copy of the plate in your lab notebook and label the tick marks as &#8216;aspirin&#8217;, &#8216;acetaminophen&#8217;, &#8216;caffeine&#8217;, and &#8216;unknown&#8217;.</div>
		</li>
		<li data-sub-note-item="2-5">
		<div class="lab-step">Now, bring your plate to the common bench. Spot solutions of aspirin, acetaminophen, caffeine, and the unknown that you were assigned on the appropriate marks using a clean capillary for each solution. <strong>Note: </strong>When you are spotting the unknown solution, don&#39;t dip the capillary too far into the solution vial to avoid clogging your capillary with insoluble material from the unknown substance.</div>
		</li>
		<li data-sub-note-item="2-6">
		<div class="lab-step">Discard the capillaries and close the vials before bringing the plate back to your fume hood. Place the plate on a paper towel to finish drying.</div>
		</li>
		<li data-sub-note-item="2-7">
		<div class="lab-step">Next, label a clean 50-mL screw-top jar &#8216;90% ethyl acetate, 10% hexane&#8217;, and bring it in a clean graduated cylinder to the solvent hood.</div>
		</li>
		<li data-sub-note-item="2-8">
		<div class="lab-step">Place 2 &#8211; 3 mL of the 90/10 solution in the jar and return to your fume hood. Confirm that the solvent level will be below the spots on the plate. Then, place a clean filter paper upright in the jar and close it tightly.</div>
		</li>
		<li data-sub-note-item="2-9">
		<div class="lab-step">Once the TLC plate is dry and the filter paper is saturated, carefully place the plate in the solvent. <strong>Note:</strong> It&#39;s essential to hold the plate level when you do this. If the solvent front is diagonal, the solvent distance traveled will be different for each spot, making it impossible to visually compare the unknown to the reference spots.</div>
		</li>
		<li data-sub-note-item="2-10">
		<div class="lab-step">Wait for the solvent front to reach 1 cm from the top of the plate. Then, retrieve the plate, mark it solvent front, and let it dry while lying flat.</div>
		</li>
		<li data-sub-note-item="2-11">
		<div class="lab-step">Once the plate is dry, visualize it with UV light and in the iodine chamber, and outline the spots in pencil.</div>
		</li>
		<li data-sub-note-item="2-12">
		<div class="lab-step">The unknown material could be a mixture of compounds, so look carefully for spots in line with all three reference spots.</div>
		</li>
		<li data-sub-note-item="2-13">
		<div class="lab-step">Then, precisely copy the spots onto the drawing of the plate in your lab notebook. Measure and record the height of the solvent front and the spots relative to the starting line.</div>
		</li>
		<li data-sub-note-item="2-14">
		<div class="lab-step">When you are finished, retrieve the solvent-saturated filter papers from the jars and let them dry in the hood.</div>
		</li>
		<li data-sub-note-item="2-15">
		<div class="lab-step">Pour leftover solvents into the standard organic waste, wash your glassware following your lab&#39;s usual methods, and put away your lab equipment.</div>
		</li>
		<li data-sub-note-item="2-16">
		<div class="lab-step">Throw out used TLC plates in the lab trash or with glass waste, depending on the backing material.</div>
		</li>
		<li data-sub-note-item="2-17">
		<div class="lab-step">Lastly, once the filters are dry, throw them out along with any remaining trash.</div>
		</li>
	</ul>
	</li>
	<li data-note-item="3" data-parent-collapse="">Results<button class="expand">Expand</button>
	<ul class="lab-items">
		<li data-sub-note-item="3-1">
		<div class="lab-step">Calculate the R<sub>f</sub> values for phenol, benzoic acid, and butyl phenyl ether eluted with 100% hexane. For each compound on the TLC plate, measure the distance from the starting line to the center of its spot. Then, divide this value by the distance from the starting line to the solvent front.</div>
		</li>
		<li data-sub-note-item="3-2">
		<div class="lab-step">Only butyl phenyl ether moved a measurable distance from the starting line, which is consistent with it being the least polar. For effective separation, the R<sub>f</sub> values should differ by 0.3 &#8211; 0.7. Thus, we can infer that phenol and benzoic acid are too polar to be separated effectively by an apolar solvent like pure hexane.</div>
		</li>
		<li data-sub-note-item="3-3">
		<div class="lab-step">Calculate the R<sub>f</sub> values for the plate that you ran in a 60/40 mixture of hexane and ethyl acetate. Ethyl acetate is a moderately polar solvent, so the compounds traveled farther and are spaced farther apart on the plate.</div>
		</li>
		<li data-sub-note-item="3-4">
		<div class="lab-step">The separation order shows that benzoic acid, the most polar compound, had the highest affinity for silica gel, whereas butyl phenyl ether, the least polar compound, had the lowest affinity for silica gel. This is consistent with the assumption that less polar compounds travel farther on polar stationary phases.</div>
		</li>
		<li data-sub-note-item="3-5">
		<div class="lab-step">Look at your third plate and calculate the R<sub>f</sub> values for aspirin, acetaminophen, and caffeine. Caffeine, the most polar complex of the three, traveled the least. Acetaminophen traveled the second farthest, while aspirin, the least polar, traveled the most.</div>
		</li>
		<li data-sub-note-item="3-6">
		<div class="lab-step">Finally, identify the compounds in your unknown material by comparing its spot or spots to the three reference spots.</div>
		</li>
	</ul>
	</li>
</ol>

Thin-Layer Chromatography: Experimental Setup and Separation - Procedure

Source: Lara Al Hariri and&#160;Ahmed Basabrain at the University of Massachusetts Amherst, MA, USA

	Effects of Mobile Phase Composition on Compound ...

Education

JoVE Lab Manual

Chemistry

Lab Manual Sub Articles

Temperature Dependence

<p>Source: Smaa Koraym at Johns Hopkins University, MD, USA</p>
<ol class="note-items">
	<li data-note-item="1" data-parent-collapse="">Temperature Dependent Decomposition of Hydrogen Peroxide<button class="expand">Expand</button>
	<p><em>We suggest that students work in pairs for this experiment. Equipment controls may vary.</em></p>
	<p>In this lab, you&#39;ll perform a decomposition reaction where a single compound breaks down into two or more simpler products. You will observe the decomposition of hydrogen peroxide into water and oxygen. This decomposition happens very slowly, so you will use iron (III) nitrate as a catalyst to lower the activation energy.</p>
	<p>During this process, iron undergoes a redox reaction and then returns to its starting oxidation state. You will be able to see this as a color change in your solution during the reaction. You&#39;ll perform the same reaction at four different temperatures and track the rate of the reaction by recording the pressure inside the flask. This way you can measure how quickly oxygen was produced in each experiment. You&#39;ll use this to calculate the reaction&#39;s activation energy.</p>
	<p><strong>Table 1. Estimate the apparent activation energy for the decompostion of hydrogen peroxide</strong></p>
	<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
		<tbody>
			<tr>
				<td><strong>Trial&#160;</strong></td>
				<td><strong>Temperature (&#8451;)</strong></td>
				<td><strong>&#916;P (kPa/s)</strong></td>
				<td><strong>1/T&#160;</strong></td>
				<td><strong>ln (&#916;P)</strong></td>
			</tr>
			<tr>
				<td>1</td>
				<td></td>
				<td></td>
				<td></td>
				<td></td>
			</tr>
			<tr>
				<td>2</td>
				<td></td>
				<td></td>
				<td></td>
				<td></td>
			</tr>
			<tr>
				<td>3</td>
				<td></td>
				<td></td>
				<td></td>
				<td></td>
			</tr>
			<tr>
				<td>4</td>
				<td></td>
				<td></td>
				<td></td>
				<td></td>
			</tr>
		</tbody>
	</table>
	<p><a href="/CDNSource/labs/51/Table_1._Estimate_the_apparent_activation_energy_of.xlsx" target="_blank">Click Here to download Table 1</a> &#160;</p>
	<ul class="lab-items">
		<li data-sub-note-item="1-1">
		<div class="lab-step">Before starting this experiment, put on a lab coat, safety glasses, and nitrile gloves.</div>
		</li>
		<li data-sub-note-item="1-2">
		<div class="lab-step">Ensure that your hot plate is off and then place a 600-mL beaker on the hot plate.</div>
		</li>
		<li data-sub-note-item="1-3">
		<div class="lab-step">Connect vacuum tubing to the barbed sidearm of a 125-mL B&#252;chner filter flask, and carefully clamp the filter flask in the 600-mL beaker so that the sidearm is just above the top of the beaker.</div>
		</li>
		<li data-sub-note-item="1-4">
		<div class="lab-step">Fill a 400-mL beaker with deionized water., and pour the water into the 600-mL beaker until the water level is about 2&#8211;3 cm below the side arm.<br />
		<strong>NOTE</strong>: Make sure that the water is not able to get into the flask or vacuum line.</div>
		</li>
		<li data-sub-note-item="1-5">
		<div class="lab-step">Immerse the thermometer in the water, clamping it in place so that the end is in contact with the outer wall of the flask. Ensure that you can read the current temperature, as well as the 40, 60, and 80 &#176;C marks.</div>
		</li>
		<li data-sub-note-item="1-6">
		<div class="lab-step">Make sure that each hole of the 2-hole adapter is fitted with a tapered locking adapter. Check that all the adapters are firmly seated, since any air that escapes around them will affect your data.</div>
		</li>
		<li data-sub-note-item="1-7">
		<div class="lab-step">Lock a 2-way stopcock into one of the stopper adapters. Then, lock one end of the flexible tubing into the second adapter with the other end into the connector of the gas pressure sensor.</div>
		</li>
		<li data-sub-note-item="1-8">
		<div class="lab-step">Turn on the data acquisition device for the gas pressure sensor and ensure that the pressure is displayed in kPa.</div>
		</li>
		<li data-sub-note-item="1-9">
		<div class="lab-step">Set the acquisition rate to two samples per second and the duration to 300 s. Then, display the pressure in real-time.</div>
		</li>
		<li data-sub-note-item="1-10">
		<div class="lab-step">Label a 400-mL beaker as &#8216;waste&#8217;, a 50-mL beaker as &#8216;0.5 M Fe(NO<sub>3</sub>)<sub>3</sub>&#8217;, and a 100-mL beaker as &#8216;3% w/w hydrogen peroxide&#8217;.</div>
		</li>
		<li data-sub-note-item="1-12">
		<div class="lab-step">Place a few paper towels on the work surface as a clean area for glassware that you will reuse. Keep a supply of paper towels on hand for later.</div>
		</li>
		<li data-sub-note-item="1-13">
		<div class="lab-step">Then, bring the 50-mL and 100-mL beakers to the stock solution area. Pour about 30 mL of 0.5 M Fe(NO<sub>3</sub>)<sub>3</sub> in the 50-mL beaker and 100 mL of 3% w/w hydrogen peroxide in the 100-mL beaker.</div>
		</li>
		<li data-sub-note-item="1-14">
		<div class="lab-step">Back at your workspace, setup a 20-mL volumetric pipette and fill it to the mark with 3% w/w hydrogen peroxide. Dispense the hydrogen peroxide into the filter flask and set the volumetric pipette aside.</div>
		</li>
		<li data-sub-note-item="1-15">
		<div class="lab-step">Adjust the thermometer as needed so that it contacts the flask below the hydrogen peroxide solution level.</div>
		</li>
		<li data-sub-note-item="1-16">
		<div class="lab-step">Fit the rubber stopper into the mouth of the filter flask, being careful not to loosen the connections to the sensor and the 2-way stopcock.</div>
		</li>
		<li data-sub-note-item="1-17">
		<div class="lab-step">Check that the 2-way stop is closed. Open the vacuum line and monitor the pressure as it decreases. This will seal the stopper in the flask. Once the pressure reaches 10 kPa, close the vacuum.</div>
		</li>
		<li data-sub-note-item="1-18">
		<div class="lab-step">Monitor the pressure for at least 1 min to confirm there are no slow leaks.<br />
		<strong>NOTE</strong>: If the pressure immediately starts rapidly increasing, there is a leak in your setup, so tighten the connections and try again until the pressure holds at 10 kPa when the vacuum line is closed.</div>
		</li>
		<li data-sub-note-item="1-19">
		<div class="lab-step">Draw 5 mL of 0.5 M Fe(NO<sub>3</sub>)<sub>3</sub> into a 20-mL syringe. Expel all air from the syringe so that it only contains the solution.</div>
		</li>
		<li data-sub-note-item="1-20">
		<div class="lab-step">Lock the syringe into the top of the 2-way stopcock. You are now ready to start the room temperature trial, so record the water temperature in your lab notebook.</div>
		</li>
		<li data-sub-note-item="1-21">
		<div class="lab-step">Start acquiring gas pressure data. Allow the device to record data for about 15 s, then open the stopcock and quickly close it once all of the Fe(NO<sub>3</sub>)<sub>3</sub> has entered the flask. The observed increase in pressure is from the evolution of oxygen gas produced by the decomposition of hydrogen peroxide.</div>
		</li>
		<li data-sub-note-item="1-22">
		<div class="lab-step">Once the data collection finishes, save your data. Then, disconnect the syringe and open the stopcock to vent the flask.</div>
		</li>
		<li data-sub-note-item="1-23">
		<div class="lab-step">Take out the rubber stopper, carefully remove the thermometer from the clamp and the flask from the beaker, and disconnect the vacuum tubing from the filter flask.</div>
		</li>
		<li data-sub-note-item="1-24">
		<div class="lab-step">Empty the flask into the waste beaker. Try not to get liquid in the side arm. Dry the outside of the flask with paper towels.</div>
		</li>
		<li data-sub-note-item="1-25">
		<div class="lab-step">Rinse the interior of the flask with deionized water and pour the rinse into the waste beaker. If any liquid gets into the side arm, remove it with paper towels.</div>
		</li>
		<li data-sub-note-item="1-26">
		<div class="lab-step">Reconnect the flask to the vacuum line and clamp it in the 600-mL beaker in contact with the thermometer.</div>
		</li>
		<li data-sub-note-item="1-27">
		<div class="lab-step">Turn on the hotplate and heat the water around the flask until the thermometer reads 80 &#176;C. Then, turn off the heat.</div>
		</li>
		<li data-sub-note-item="1-28">
		<div class="lab-step">Then, add 20 mL of 3% w/w hydrogen peroxide to the flask.</div>
		</li>
		<li data-sub-note-item="1-29">
		<div class="lab-step">Dry the stopper with paper towels. Do this after each trial to ensure that the stopper has a snug fit in the neck of the beaker and fit the stopper in the flask.</div>
		</li>
		<li data-sub-note-item="1-30">
		<div class="lab-step">Check that the stopcock is closed and evacuate the flask to about 10 kPa. Then, close the vacuum and confirm that there are no leaks.</div>
		</li>
		<li data-sub-note-item="1-31">
		<div class="lab-step">Draw 5 mL Fe(NO<sub>3</sub>)<sub>3</sub> solution into the syringe, expel the air from the syringe, and connect it to the stopcock.</div>
		</li>
		<li data-sub-note-item="1-32">
		<div class="lab-step">Record the temperature shown on the thermometer in your lab notebook, then, start collecting data.</div>
		</li>
		<li data-sub-note-item="1-33">
		<div class="lab-step">Wait about 15 s and introduce the Fe(NO<sub>3</sub>)<sub>3</sub> solution in the same way as before.<br />
		<strong>NOTE:</strong> As the pressure approaches 150 kPa, the stopper may pop off!</div>
		</li>
		<li data-sub-note-item="1-34">
		<div class="lab-step">After data collection finishes, save the data, vent and clean the filter flask, and set up for the third trial.</div>
		</li>
		<li data-sub-note-item="1-35">
		<div class="lab-step">Fill the 400-mL beaker with ice and add some to cool the water to about 60 &#176;C.</div>
		</li>
		<li data-sub-note-item="1-36">
		<div class="lab-step">Perform the third trial in the same way as before. Remember to dry the stopper and the inside of the flask and to record the temperature prior to data collection.</div>
		</li>
		<li data-sub-note-item="1-37">
		<div class="lab-step">After the data is collected for that trial, cool the water to about 40 &#176;C and perform the fourth trial.</div>
		</li>
		<li data-sub-note-item="1-38">
		<div class="lab-step">Once you have finished all four trials, empty the filter flask and rinse it into the waste beaker one last time.</div>
		</li>
		<li data-sub-note-item="1-39">
		<div class="lab-step">Dispose of any excess 0.5 M Fe(NO<sub>3</sub>)<sub>3</sub> and the contents of the waste beaker in a container labeled for aqueous iron waste.</div>
		</li>
		<li data-sub-note-item="1-40">
		<div class="lab-step">Then, disassemble the apparatus and pour the leftover ice, water, and hydrogen peroxide in the sink. Wash your glassware following your lab&#39;s standard protocol.</div>
		</li>
	</ul>
	</li>
	<li data-note-item="1" data-parent-collapse="">Results<button class="expand">Expand</button>
	<p>The decomposition of hydrogen peroxide with iron is a complex, multi-step process that we can&#39;t easily describe in a single equation. However, we can estimate the apparent activation energy from the rate of oxygen production and compare it to the apparent activation energy of the un-catalyzed process.</p>
	<ul class="lab-items">
		<li data-sub-note-item="2-1">
		<div class="lab-step">Find the rate of pressure change, which is directly proportional to the rate of oxygen production. For each experiment, make a plot of pressure with respect to time and find the point where the reaction started.</div>
		</li>
		<li data-sub-note-item="2-2">
		<div class="lab-step">Identify the maximum pressure attained and determine the slope between the two points.</div>
		</li>
		<li data-sub-note-item="2-3">
		<div class="lab-step">Once you&#39;ve determined the slopes and the corresponding temperatures in Kelvin for all of the temperatures, use the Arrhenius equation to estimate the apparent activation energy of this reaction.</div>
		</li>
		<li data-sub-note-item="2-4">
		<div class="lab-step">Take the reciprocal of the temperatures in Kelvin and the natural log of the rate of change in pressure. Remember that the rate constant k is essentially equal to the change in pressure.</div>
		</li>
		<li data-sub-note-item="2-5">
		<div class="lab-step">Make an Arrhenius plot and find the slope of the line.</div>
		</li>
		<li data-sub-note-item="2-6">
		<div class="lab-step">The slope is equal to the negative activation energy over the universal gas constant, so multiply the slope by the negative universal gas constant to get the apparent activation energy of the iron catalyzed decomposition reaction. You&#39;ll see a value in the range of 35&#8211;60 kJ/mol because the iron catalyst made the decomposition take less energy. The apparent activation energy of the uncatalyzed decomposition of hydrogen peroxide is around 78 &#8211; 88 kJ/mol.</div>
		</li>
	</ul>
	</li>
</ol>

Temperature Dependence and Arrhenius Equation - Procedure

Source: Smaa Koraym at Johns Hopkins University, MD, USA

	Temperature Dependent Decomposition of Hydrogen PeroxideExpand
	We suggest that students work ...

Galvanic Cells

Galvanic Cells: Construction and Reduction Potential Measurement - Procedure

Source: Smaa Koraym at Johns Hopkins University, MD, USA

	Constructing a Lead-Copper Galvanic CellExpand
	In this experiment, you will construct a ...

Electrolytic Cells

<p>Source: Smaa Koraym at Johns Hopkins University, MD, USA</p>
<ol class="note-items">
	<li data-note-item="1" data-parent-collapse="">Electrolytic Cells<button class="expand">Expand</button>
	<p>In this experiment, you will be assembling an electrolytic cell to perform electroplating. An electrolytic cell contains an anode and cathode connected by a power source, which drives the nonspontaneous redox reaction in the cell. When the anode material is oxidized, the ions travel to the cathode, where they are reduced and plated.</p>
	<p>In the case of this experiment, the electrolytic cell contains a copper anode and a brass key for the cathode. Oxidation of the anode creates copper(2+) ions, which are then reduced on the brass key, forming a thin plating of solid copper.</p>
	<ul class="lab-items">
		<li data-sub-note-item="1-3">
		<div class="lab-step">Put on gloves, safety goggles, and a lab apron, which must be worn at all times.</div>
		</li>
		<li data-sub-note-item="1-4">
		<div class="lab-step">Obtain one 10-ohm resistor, one copper sheet electrode, one copper wire, and one piece of emery paper.</div>
		</li>
		<li data-sub-note-item="1-5">
		<div class="lab-step">Use the emery paper to polish the brass key and the copper sheet. Rinse the key and the copper electrode with water. The brass key will be the cathode, and the copper sheet will be the anode.</div>
		</li>
		<li data-sub-note-item="1-6">
		<div class="lab-step">Take one 50-mL beaker and a watch glass to the instructor&#39;s hood and obtain approximately 20 mL of acetone. Place the watch glass on top of the beaker to prevent evaporation and bring it back to your bench.</div>
		</li>
		<li data-sub-note-item="1-7">
		<div class="lab-step">Label the other 50-mL beaker as &#39;organic waste&#39;.</div>
		</li>
		<li data-sub-note-item="1-8">
		<div class="lab-step">Use a disposable pipette to rinse the copper electrode and key with acetone. Collect excess in the waste beaker. Place the key and copper electrode on a paper towel to dry.</div>
		</li>
		<li data-sub-note-item="1-9">
		<div class="lab-step">Measure the initial masses of the key and copper electrode and record their values in your lab notebook.
		<h4><strong>Table 1: Moles of Cu Transferred</strong></h4>
		<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
			<tbody>
				<tr>
					<td></td>
					<td style="text-align: center;"><strong>Mass of key (g) </strong></td>
					<td style="text-align: center;"><strong>Change in mass (g) </strong></td>
					<td style="text-align: center;"><strong>Moles Cu plated </strong></td>
					<td style="text-align: center;"><strong>Mass of Cu electrode (g) </strong></td>
					<td style="text-align: center;"><strong>Change in mass (g) </strong></td>
					<td style="text-align: center;"><strong>Moles Cu lost </strong></td>
				</tr>
				<tr>
					<td style="text-align: center;">0 min</td>
					<td></td>
					<td></td>
					<td></td>
					<td></td>
					<td></td>
					<td></td>
				</tr>
				<tr>
					<td style="text-align: center;">5 min</td>
					<td></td>
					<td></td>
					<td></td>
					<td></td>
					<td></td>
					<td></td>
				</tr>
				<tr>
					<td style="text-align: center;">1<sup>st</sup> 30 min</td>
					<td></td>
					<td></td>
					<td></td>
					<td></td>
					<td></td>
					<td></td>
				</tr>
				<tr>
					<td style="text-align: center;">2<sup>nd</sup> 30 min</td>
					<td></td>
					<td></td>
					<td></td>
					<td></td>
					<td></td>
					<td></td>
				</tr>
			</tbody>
		</table>
		<a href="https://www-jove-com.remotexs.ntu.edu.sg/CDNSource/lm/labs/53/Table_1._Moles_of_Cu_Transferred.xlsx" target="_blank">Click Here to download Table 1</a></div>
		</li>
		<li data-sub-note-item="1-10">
		<div class="lab-step">Set up the electrolytic cell. Use one electrical wire with alligator clips to connect the negative terminal on the battery to the current probe. Wrap the 10-ohm resistor around the other end of the current probe.</div>
		</li>
		<li data-sub-note-item="1-11">
		<div class="lab-step">Use another wire to connect the positive terminal on the battery to the copper electrode.</div>
		</li>
		<li data-sub-note-item="1-12">
		<div class="lab-step">Wrap the copper wire around the brass key and use an electrical wire to connect the resistor on the current probe to the copper wire on the brass key.</div>
		</li>
		<li data-sub-note-item="1-13">
		<div class="lab-step">Obtain about 200 mL of concentrated copper sulfate in a 250-mL beaker. The amount does not need to be exact. Bring the beaker back to your bench and set it on the stir plate. Carefully, place a magnetic stir bar in the beaker and turn the stir setting on to high.</div>
		</li>
		<li data-sub-note-item="1-14">
		<div class="lab-step">Connect the data acquisition system to the current probe and allow the data acquisition system to sense the probe. It should return a reading in amps. Change the rate to two scans per min and the duration to 5 min.</div>
		</li>
		<li data-sub-note-item="1-15">
		<div class="lab-step">Place the brass key in the beaker. Hang it off the side of the beaker so that the key is almost completely submerged but does not touch the magnetic stir bar. <strong>Note: </strong>The copper wire should not be in the solution.</div>
		</li>
		<li data-sub-note-item="1-16">
		<div class="lab-step">Place the copper electrode in the beaker, bending it so that it hangs over the edge of the beaker. Ensure that it is not touching the stir bar or the brass key.</div>
		</li>
		<li data-sub-note-item="1-17">
		<div class="lab-step">Start the scan and let it collect data for 5 min. This will act as a test run to make sure plating occurs on the brass key.</div>
		</li>
		<li data-sub-note-item="1-18">
		<div class="lab-step">After the scan ends, remove the key and copper electrode. The copper plating on the brass key should be evident. If it is not visible, check your circuit to make sure that the battery is connected in the correct orientation.</div>
		</li>
		<li data-sub-note-item="1-19">
		<div class="lab-step">Once you have completed a successful 5-min plating, disconnect the key and copper electrode from the wiring. Remove the copper wire from the key and rinse the key and copper electrode with water and then acetone. Lay them on a paper towel to dry.</div>
		</li>
		<li data-sub-note-item="1-20">
		<div class="lab-step">Measure the masses of the key and the copper electrode and record them in your lab notebook.</div>
		</li>
		<li data-sub-note-item="1-21">
		<div class="lab-step">Reconnect the key and copper electrode and place them back in the solution like before. Restart the run on the data acquisition system, making sure that the rate is set to two scans per min. With the duration set to 30 min, allow the electroplating process to run.</div>
		</li>
		<li data-sub-note-item="1-22">
		<div class="lab-step">After the scan is complete, remove the electrode and key from the solution, disconnect them from the wires, and rinse them with water and then acetone. Allow them to dry on a paper towel, then weigh them and record their masses in your notebook.</div>
		</li>
		<li data-sub-note-item="1-23">
		<div class="lab-step">Reassemble the setup and repeat the electroplating for a second 30-min trial. When the scan is complete, remove the key and copper electrode, rinse them like before, and record their mass.</div>
		</li>
		<li data-sub-note-item="1-24">
		<div class="lab-step">Save the recorded data as a text file on a flash drive.</div>
		</li>
		<li data-sub-note-item="1-25">
		<div class="lab-step">To clean up, turn the stir setting off and remove the stir bar from the solution using forceps. Dispose of the acetone in the organic waste container provided by your instructor. The copper sulfate solution can be reused for future labs, so pour the concentrated copper sulfate back into the carboy.</div>
		</li>
		<li data-sub-note-item="1-26">
		<div class="lab-step">Rinse all of your glassware with water and return it to the instructor.</div>
		</li>
	</ul>
	</li>
	<li data-note-item="2" data-parent-collapse="">Results<button class="expand">Expand</button>
	<ul class="lab-items">
		<li data-sub-note-item="2-1">
		<div class="lab-step">First, calculate the total charge Q that flowed through the system during the different trials. One coulomb is the quantity of electricity flowing through a wire carrying 1 ampere of current in 1 s, thus Q = I x t.</div>
		</li>
		<li data-sub-note-item="2-2">
		<div class="lab-step">Review the current probe data for the first 5-min electroplating trial.<strong> Note: </strong>If the current dropped to 0 or close to 0 at the end of the trial, then the electrons stopped flowing from the copper electrode to the key. Only use the values recorded before the drop in current occurred. If the current drop to 0 early in the plating process, repeat the trial.</div>
		</li>
		<li data-sub-note-item="2-3">
		<div class="lab-step">The current varies over time, so calculate the average current, I.
		<h4><strong>Table 2: Moles of Electrons Transferred</strong></h4>
		<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
			<tbody>
				<tr>
					<td style="text-align: center;"><strong>Trial</strong></td>
					<td style="text-align: center;"><strong>Time (s) </strong></td>
					<td style="text-align: center;"><strong>Current<sub>average</sub> (A) </strong></td>
					<td style="text-align: center;"><strong>Total charge (C) </strong></td>
					<td style="text-align: center;"><strong>Moles (e?) </strong></td>
				</tr>
				<tr>
					<td style="text-align: center;">5 min</td>
					<td></td>
					<td></td>
					<td></td>
					<td></td>
				</tr>
				<tr>
					<td style="text-align: center;">1<sup>st</sup> 30 min</td>
					<td></td>
					<td></td>
					<td></td>
					<td></td>
				</tr>
				<tr>
					<td style="text-align: center;">2<sup>nd</sup> 30 min</td>
					<td></td>
					<td></td>
					<td></td>
					<td></td>
				</tr>
				<tr>
					<td style="text-align: center;">Total</td>
					<td></td>
					<td></td>
					<td></td>
					<td></td>
				</tr>
			</tbody>
		</table>
		<a href="https://www-jove-com.remotexs.ntu.edu.sg/CDNSource/lm/labs/53/Table_2._Moles_of_Electrons_Transferred.xlsx" target="_blank">Click Here to download Table 2</a></div>
		</li>
		<li data-sub-note-item="2-4">
		<div class="lab-step">Use the average current over the total time to determine the total charge transferred during electroplating, Q. About 20 coulombs were transferred during the 5-min trial, and between 110 and 120 coulombs were transferred during each 30-min trial. The total charge transferred over the entire 65-min electroplating process was about 250 coulombs.</div>
		</li>
		<li data-sub-note-item="2-5">
		<div class="lab-step">Use the total charge transferred to calculate the moles of electrons plated during each trial. The moles of electrons plated equals the total charge transferred divided by the Faraday constant. The Faraday constant represents the magnitude of electric charge per mole of electrons and is determined by multiplying the charge of an electron by Avogadro&#39;s constant.</div>
		</li>
		<li data-sub-note-item="2-6">
		<div class="lab-step">Determine the moles of electrons that were transferred for the 5-min and 30-min trials.</div>
		</li>
		<li data-sub-note-item="2-7">
		<div class="lab-step">Calculate the moles of copper that were plated on the key after each trial. Similarly, determine the moles of copper lost from the copper sheet electrode.</div>
		</li>
		<li data-sub-note-item="2-8">
		<div class="lab-step">Compare the moles of copper that were lost from the electrode to the amount that was plated to the brass key. The numbers should be close.</div>
		</li>
		<li data-sub-note-item="2-9">
		<div class="lab-step">Check to see if these values match what is expected based on the number of moles of electrons that were transferred. The reduction of 1 mole of copper(2+) ions requires 2 moles of electrons to produce 1 mole of solid copper.</div>
		</li>
	</ul>
	</li>
</ol>

Electrolytic Cells: Construction and Electroplating - Procedure

Source: Smaa Koraym at Johns Hopkins University, MD, USA

	Electrolytic CellsExpand
	In this experiment, you will be assembling an electrolytic cell to ...

Proper Lab Notebook Keeping

<p>Source: Lara Al Hariri and Ahmed Basabrain at the University of Massachusetts Amherst, MA, USA</p>
<ol class="note-items">
	<li data-note-item="1" data-parent-collapse="">Nitration of Methyl Benzoate<button class="expand">Expand</button>
	<p>From this lab onward, you&#39;ll keep a record of every experiment in your lab notebook. This starts with preparing your lab notebook before you come to the lab. This preparation, which is often called a pre-lab, typically includes balanced chemical equations for the reactions, important properties of the reagents and products, and a step-by-step procedure for the lab in your own words. In this lab, you will perform the nitration of methyl benzoate, an aromatic ester that will undergo an electrophilic aromatic substitution to make methyl 3-nitrobenzoate.</p>
	<p>First, you&#39;ll mix aqueous sulfuric acid and nitric acid to make positive nitronium ions. In a separate container, you&#39;ll mix sulfuric acid with methyl benzoate to protonate the ester group. The way the charge is distributed makes the ring most reactive towards positive ions at the carbons one step away from the ester. This is known as the meta position in di-substituted benzene rings. When you combine the solutions, nitronium adds to methyl benzoate at one of those carbons. Then, the benzene ring and the ester group lose their extra protons to make methyl 3-nitrobenzoate.</p>
	<p>After the reaction, you&#39;ll dilute the mixture with water from melted ice and collect the insoluble product by vacuum filtration. At the end, you&#39;ll estimate the yield of your reaction based on the starting volume of methyl benzoate and the mass of the product.</p>
	<p>During the experiment, make sure to write down what you did and your observations as they happen. An accurate record is essential for reproducing your results and identifying sources of experimental error. Always catch up on your lab notebook before starting the next step.</p>
	<ul class="lab-items">
		<li data-sub-note-item="1-5">
		<div class="lab-step">Before you start the experiment, put on a lab coat, safety glasses, and nitrile gloves. Make sure to review the safety information in your pre-lab.</div>
		</li>
		<li data-sub-note-item="1-6">
		<div class="lab-step">Fill a 100-mL beaker about halfway with crushed ice. <strong>Note:</strong> The reactions in this lab are exothermic, so use ice baths to keep the solutions cool.</div>
		</li>
		<li data-sub-note-item="1-7">
		<div class="lab-step">Prepare the mixture of nitric acid and sulfuric acid. In your fume hood, clamp a 20-mL test tube upright in the ice bath. Label two 10-mL graduated cylinders &#8216;nitric acid&#8217; and &#8216;sulfuric acid&#8217;.</div>
		</li>
		<li data-sub-note-item="1-8">
		<div class="lab-step">Measure 2 mL of concentrated sulfuric acid. If you add too much, use a pipette to transfer the excess to a small beaker labeled &#8216;sulfuric acid waste&#8217;.</div>
		</li>
		<li data-sub-note-item="1-9">
		<div class="lab-step">Record the volume of sulfuric acid in your lab notebook as the volume used in the mixture of acids (Step 1). Pour the sulfuric acid into the test tube.
		<h4><strong>Table 1: Nitration of methyl benzoate</strong></h4>
		<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
			<tbody>
				<tr>
					<td><strong>Step 1: Make positive nitronium ions</strong></td>
					<td style="text-align: center;">&#160; &#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
				</tr>
				<tr>
					<td>Volume of sulfuric acid (mL)</td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>Volume of nitiric acid (mL)</td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>Time of stirring (min)</td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td><strong>Step 2: Protonate ester on methyl benzoate</strong></td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>Volume of sulfuric acid (mL)</td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>Volume of methyl benzoate (mL)</td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td><strong>Step 3: Combine solutions to form 3-methyl benzoate </strong></td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>Time of addition (min)</td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>Reaction time on ice (min)</td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>Reaction time at RT (min)</td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>Yield of methyl 3-nitrobenzoate (g)</td>
					<td style="text-align: center;"></td>
				</tr>
			</tbody>
		</table>
		<a href="https://www-jove-com.remotexs.ntu.edu.sg/CDNSource/lm/labs/55/Table_1._Nitration_of_methyl_benzoate.xlsx" target="_blank">Click Here to download Table 1</a></div>
		</li>
		<li data-sub-note-item="1-10">
		<div class="lab-step">Use the other graduated cylinder to measure 2 mL of concentrated nitric acid and pipette any extra nitric acid into a separate waste beaker. Record the volume of nitric acid in the graduated cylinder before pouring it into the test tube.</div>
		</li>
		<li data-sub-note-item="1-11">
		<div class="lab-step">Stir the solution for 2 min with a glass rod. Don&#39;t remove the solution from the ice bath. When you&#39;re done, record for how long you stirred the solution.</div>
		</li>
		<li data-sub-note-item="1-12">
		<div class="lab-step">Fill a 400-mL beaker about 3/4 full with ice and place it by an available lab stand in your fume hood. Clamp a clean 25-mL Erlenmeyer flask in this ice bath.</div>
		</li>
		<li data-sub-note-item="1-13">
		<div class="lab-step">Prepare the mixture of sulfuric acid and methyl benzoate (Step 2). Measure another 2 mL of sulfuric acid with the corresponding graduated cylinder. Record the volume and carefully pour it into the Erlenmeyer flask.</div>
		</li>
		<li data-sub-note-item="1-14">
		<div class="lab-step">Bring a clean 10-mL graduated cylinder, a pipette, and a small beaker to the solvent hood.</div>
		</li>
		<li data-sub-note-item="1-15">
		<div class="lab-step">Carefully measure 1.1 mL of methyl benzoate and pipette any excess into the beaker for disposal in the organic waste. Remember to record the volume of methyl benzoate before pouring it into the flask of sulfuric acid.</div>
		</li>
		<li data-sub-note-item="1-16">
		<div class="lab-step">Place a thermometer in the flask and wait for the solution to cool to between 10 &#176;C and 15 &#176;C.</div>
		</li>
		<li data-sub-note-item="1-17">
		<div class="lab-step">Once the methyl benzoate solution is below 15 &#176;C, get a clean Pasteur pipette. Note the time in your lab notebook as the start of the solution addition and transfer the glass rod from the test tube to the Erlenmeyer flask (Step 3).</div>
		</li>
		<li data-sub-note-item="1-18">
		<div class="lab-step">At a rate of 2 - 3 drops per min, add the mixed acids to the methyl benzoate solution and stir the mixture with the glass rod. Monitor the temperature of the solution throughout the addition process.<strong> Note: </strong>If the solution reaches 15 &#176;C, wait until the temperature has dropped below 15 &#176;C before adding more of the acid mixture. The addition process will take about 10 to 15 min.</div>
		</li>
		<li data-sub-note-item="1-19">
		<div class="lab-step">Once you&#39;ve finished, note the time in your lab notebook and leave the flask in the ice bath for another 10 min while the mixture reacts.</div>
		</li>
		<li data-sub-note-item="1-20">
		<div class="lab-step">Obtain about 20 mL of deionized water in a graduated cylinder. Fill a 600-mL beaker with ice and water and place the graduated cylinder in it to chill.</div>
		</li>
		<li data-sub-note-item="1-21">
		<div class="lab-step">Once the solution has reacted for 10 min on ice, remove the flask from the ice bath, record the time in your lab notebook, and let the solution continue reacting at room temperature for another 10 to 15 min.</div>
		</li>
		<li data-sub-note-item="1-22">
		<div class="lab-step">While you wait, fill a 100-mL beaker with about 25 mL of fresh ice. When the solution has reacted at room temperature for at least 10 min, record the time as the end of the reaction and pour the mixture into the 100-mL beaker containing ice. <strong>Note:</strong> Using ice lets you dilute the solution without heating it up too much because the heat from the reaction of sulfuric acid and water is expended on melting the ice.</div>
		</li>
		<li data-sub-note-item="1-23">
		<div class="lab-step">Wait for the ice to melt, which usually takes about 20 min. Remember to write down your observations, such as how long it took for the solid product to appear and what the precipitate looks like.</div>
		</li>
		<li data-sub-note-item="1-24">
		<div class="lab-step">Once the ice has melted completely, set up for vacuum filtration using a 250-mL filter flask. Place a filter paper in the funnel and use a clean pipette to wet the filter paper with cold water from the graduated cylinder.</div>
		</li>
		<li data-sub-note-item="1-25">
		<div class="lab-step">Open the flask to vacuum and pour your product mixture into the funnel. Rinse the remaining solid into the funnel with cold water. Then, pour the rest of the cold water into the funnel to wash the solid.</div>
		</li>
		<li data-sub-note-item="1-26">
		<div class="lab-step">Once no more liquid is dripping from the funnel into the flask, close the vacuum and break the vacuum seal. Carefully place the funnel in a beaker so that your product can dry overnight.</div>
		</li>
		<li data-sub-note-item="1-27">
		<div class="lab-step">Dispose of the filtrate in the aqueous acidic waste container. Pour the ice baths into the sink and wash your glassware using your usual methods. Dispose of used Pasteur pipettes in the glass waste container, neutralize and clean up any spilled acid, and throw out paper waste in the lab trash.</div>
		</li>
		<li data-sub-note-item="1-28">
		<div class="lab-step">Calculate the theoretical yield of methyl 3-nitrobenzoate in g.
		<h4><strong>Table 2: Product yield of methyl 3-methyl benzoate</strong></h4>
		<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
			<tbody>
				<tr>
					<td>Volume of methyl benzoate (mL)</td>
					<td style="text-align: center;">&#160; &#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
				</tr>
				<tr>
					<td>Density of methyl benzoate (g/cm3)</td>
					<td style="text-align: center;">1.08</td>
				</tr>
				<tr>
					<td>Molecular weight of methyl benzoate (g/mol)</td>
					<td style="text-align: center;">136.15</td>
				</tr>
				<tr>
					<td>Amount of methyl benzoate (mol)</td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>Molecular weight of methyl 3-nitrobenzoate (g/mol)</td>
					<td style="text-align: center;">181.15</td>
				</tr>
				<tr>
					<td>Theoretical yield of methyl 3-nitrobenzoate (g)</td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>Actual yield of methyl 3-nitrobenzoate (g)</td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>Actual yield of methyl 3-nitrobenzoate (mol)</td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>Percent yield</td>
					<td style="text-align: center;"></td>
				</tr>
			</tbody>
		</table>
		<a href="https://www-jove-com.remotexs.ntu.edu.sg/CDNSource/lm/labs/55/Table_2._Product_yield_of_methyl_3-methylbenzoate.xlsx" target="_blank">Click Here to download Table 2</a></div>
		</li>
		<li data-sub-note-item="1-29">
		<div class="lab-step">The next day, check on your product. If it seems dry, bring your funnel to a balance to weigh your product.<strong>Note:</strong> If it seems wet, leave it to dry some more.</div>
		</li>
		<li data-sub-note-item="1-30">
		<div class="lab-step">Tare a weighing boat and use a small spatula to carefully transfer the product from the filter paper to the weighing boat.</div>
		</li>
		<li data-sub-note-item="1-31">
		<div class="lab-step">Compare the mass of your product to the theoretical yield. If the mass reading is stable and less than the theoretical yield, you can assume that the product is dry. Record this mass in your lab notebook.</div>
		</li>
		<li data-sub-note-item="1-32">
		<div class="lab-step">Throw out the solid product and the filter paper in the lab trash and wash the B&#252;chner funnel.</div>
		</li>
	</ul>
	</li>
	<li data-note-item="2" data-parent-collapse="">Results<button class="expand">Expand</button>
	<ul class="lab-items">
		<li data-sub-note-item="2-1">
		<div class="lab-step">Calculate the yield of your reaction. From the theoretical maximum yield, convert the mass of your product to moles.</div>
		</li>
		<li data-sub-note-item="2-2">
		<div class="lab-step">Divide that amount by the maximum yield and multiply it by 100. The total nitrated product yield, which includes minor products, is usually about 85%.</div>
		</li>
	</ul>
	</li>
</ol>

Source: Lara Al Hariri and Ahmed Basabrain at the University of Massachusetts Amherst, MA, USA

	Nitration of Methyl BenzoateExpand
	From this lab onward, ...

Scientific Measurement and Lab Skills

Two-Dimensional Graphs, Curve Fitting, and Standard Deviation - Procedure

Source: Smaa Koraym at Johns Hopkins University, MD, USA

	Measuring Diameter and CircumferenceExpand
	Error can be introduced in any experiment, even ...

Acid and Base Concentrations

<p>Source: Smaa Koraym at Johns Hopkins University, MD, USA</p>
<ol class="note-items">
	<li data-note-item="1" data-parent-collapse="">Preparation of ~0.1 M NaOH<button class="expand">Expand</button>
	<p>In the first part of the lab, you will use a 50% w/w solution of NaOH to prepare 500 mL of ~0.1 M. The 50% w/w NaOH is indicative of its weight ratio. For example, if the instructor prepared 150 mL of the 50% w/w NaOH solution, then 150 g of NaOH was dissolved in 150 g of water, and the total weight of the solution is 300 g.</p>
	<ul class="lab-items">
		<li data-sub-note-item="1-2">
		<div class="lab-step">To begin, put on the appropriate personal protective equipment, including gloves, chemical splash goggles, and a lab coat.</div>
		</li>
		<li data-sub-note-item="1-3">
		<div class="lab-step">Calculate the molarity of 50% w/w NaOH, which is M<sub>1</sub> in the dilution formula.<strong> Note: </strong>The density of 50% w/w NaOH is 1.53 g/mL, and the molar mass of NaOH is 39.998 g/mole.
		<h4>Table 1. Preparation of 0.1 M NaOH from 50% w/w NaOH</h4>
		<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
			<tbody>
				<tr>
					<td>Density of 50% w/w stock solution</td>
					<td style="text-align: center;">1.53 g/mL</td>
				</tr>
				<tr>
					<td>Molar mass<sub>NaOH</sub></td>
					<td style="text-align: center;">39.998 g/mol</td>
				</tr>
				<tr>
					<td>Mass of NaOH in 50% w/w stock solution (mg)</td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>Total mass of 50% w/w stock solution</td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>Volume of 50% w/w stock solution (mL)</td>
					<td></td>
				</tr>
				<tr>
					<td>Moles of NaOH in 50% w/w solution (mol)</td>
					<td></td>
				</tr>
				<tr>
					<td>Molarity of 50% w/w stock solution (M<sub>1</sub>)</td>
					<td></td>
				</tr>
				<tr>
					<td>Volume of 50% w/w solution needed (V<sub>1</sub>)</td>
					<td></td>
				</tr>
			</tbody>
		</table>
		<a href="https://www-jove-com.remotexs.ntu.edu.sg/CDNSource/lm/labs/44/Table_1._Preparation_of_0.1_M_NaOH.xlsx" target="_blank">Click Here to download Table 1</a></div>
		</li>
		<li data-sub-note-item="1-4">
		<div class="lab-step">Use the dilution formula to determine the volume of 50% w/w NaOH needed to prepare 500 mL of ~0.1 M NaOH.</div>
		</li>
		<li data-sub-note-item="1-5">
		<div class="lab-step">Label the 500 mL polyethylene bottle as &#8216;~0.1 M NaOH&#8217;.</div>
		</li>
		<li data-sub-note-item="1-6">
		<div class="lab-step">Adjust the volume on a 1-mL pipette to the value calculated and use it to transfer the 50% w/w NaOH to the polyethylene bottle.</div>
		</li>
		<li data-sub-note-item="1-7">
		<div class="lab-step">Use a 100-mL graduated cylinder to measure the amount of water and pour it into the bottle containing the NaOH. Cap the bottle and invert it several times to mix the solution. <strong>Note:</strong> You will need 500 mL minus the calculated volume of NaOH.</div>
		</li>
	</ul>
	</li>
	<li data-note-item="2" data-parent-collapse="">Standardization of 0.1 M NaOH<button class="expand">Expand</button>
	<p>After you have prepared 0.1 M NaOH, determine its exact concentration, or standardize it, using the acid-base titration method. In this technique, a base like NaOH is slowly added to an acid like potassium hydrogen phthalate (KHP). The chemical reaction that takes place in the flask is a neutralization reaction. In this neutralization reaction, one mole of base neutralizes one mole of acid, resulting in salt and water. This reaction is performed in the presence of the indicator phenolphthalein, which is colorless at the start of the reaction when the pH is acidic. The indicator turns pink as soon as enough NaOH is added to the flask to make the pH basic.</p>
	<ul class="lab-items">
		<li data-sub-note-item="2-2">
		<div class="lab-step">Label the three Erlenmeyer flasks as &#8216;A&#8217;, &#8216;B&#8217;, and &#8216;C&#8217;.</div>
		</li>
		<li data-sub-note-item="2-3">
		<div class="lab-step">Weigh 0.5 &#8211; 0.7 g of KHP for each of the flasks and record the mass for each. <strong>Note: </strong>Try to measure the same mass of KHP for all three flasks.
		<h4><strong>Table 2. Standardization of NaOH</strong></h4>
		<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
			<tbody>
				<tr>
					<td><strong>Molar mass<sub>KHP</sub> = 204.23 g/mol </strong></td>
					<td style="text-align: center;"><strong>Flask A</strong></td>
					<td style="text-align: center;"><strong>Flask B</strong></td>
					<td style="text-align: center;"><strong>Flask C</strong></td>
				</tr>
				<tr>
					<td>Mass of KHP (g)</td>
					<td></td>
					<td></td>
					<td></td>
				</tr>
				<tr>
					<td>Volume<sub>initial NaOH</sub>(mL)</td>
					<td></td>
					<td></td>
					<td></td>
				</tr>
				<tr>
					<td>Volume<sub>final NaOH</sub> (mL)</td>
					<td></td>
					<td></td>
					<td></td>
				</tr>
				<tr>
					<td>Volume<sub>NaOH</sub> (mL)</td>
					<td></td>
					<td></td>
					<td></td>
				</tr>
				<tr>
					<td>Moles of KHP</td>
					<td></td>
					<td></td>
					<td></td>
				</tr>
				<tr>
					<td>Moles of NaOH</td>
					<td></td>
					<td></td>
					<td></td>
				</tr>
				<tr>
					<td>Molarity of NaOH</td>
					<td></td>
					<td></td>
					<td></td>
				</tr>
				<tr>
					<td>Average molarity</td>
					<td></td>
				</tr>
				<tr>
					<td>Standard deviation</td>
					<td></td>
				</tr>
			</tbody>
		</table>
		<a href="https://www-jove-com.remotexs.ntu.edu.sg/CDNSource/lm/labs/44/Table_2._Standardization_of_NaOH.xlsx" target="_blank">Click Here to download Table 2</a></div>
		</li>
		<li data-sub-note-item="2-4">
		<div class="lab-step">Add 50 mL of deionized water to each flask, and use a glass stirring rod to stir the solutions until they appear homogeneous.</div>
		</li>
		<li data-sub-note-item="2-5">
		<div class="lab-step">Add 2-3 drops of phenolphthalein to each of the three flasks.</div>
		</li>
		<li data-sub-note-item="2-6">
		<div class="lab-step">Set up the titration apparatus. Attach the burette clamp to the ring stand, and securely clamp the 50-mL glass burette to it. Make sure the valve of the burette is closed.</div>
		</li>
		<li data-sub-note-item="2-7">
		<div class="lab-step">Label a 400-mL beaker as &#8216;waste&#8217; and place it under the burette. Rinse the burette by pouring about 5 mL of 0.1 M NaOH into the burette. Open the burette valve to allow the NaOH to flow into the beaker.</div>
		</li>
		<li data-sub-note-item="2-8">
		<div class="lab-step">Close the valve and fill the burette with slightly more than 50 mL of NaOH. Release any air bubbles present in the tip of the burette by opening and closing the burette valve. Record the starting volume of NaOH.</div>
		</li>
		<li data-sub-note-item="2-9">
		<div class="lab-step">Place flask A below the tip of the burette, and titrate the solution using 1 mL volumes of NaOH. Swirl the solution after each addition. Continue adding 1 mL volumes to the flask until the pink color persists. This is considered the endpoint. Record the volume of 0.1 M NaOH added to achieve the endpoint.</div>
		</li>
		<li data-sub-note-item="2-10">
		<div class="lab-step">Repeat the titration for flasks B and C. This data will be used to calculate the actual concentration of NaOH.</div>
		</li>
	</ul>
	</li>
	<li data-note-item="3" data-parent-collapse="">Titration of phosphoric acid<button class="expand">Expand</button>
	<p>In this experiment, we will determine two of the three pKa values for the triprotic acid, phosphoric acid, using acid-base titration. In this neutralization reaction, phosphoric acid reacts with NaOH to form water and the salt, sodium phosphate.</p>
	<ul class="lab-items">
		<li data-sub-note-item="3-2">
		<div class="lab-step">Attach the drop counter to the ring stand, below the burette clamp. Secure the plastic burette so that its tip is just above the drop counter.</div>
		</li>
		<li data-sub-note-item="3-3">
		<div class="lab-step">Connect the drop counter to the data acquisition system, and make sure the two valves of the plastic burette are both in the closed position.</div>
		</li>
		<li data-sub-note-item="3-4">
		<div class="lab-step">Place the waste container below the burette and pour a few mL of 0.1 M NaOH into the burette. Open both valves to drain the NaOH into the waste beaker. Then close the valves.</div>
		</li>
		<li data-sub-note-item="3-5">
		<div class="lab-step">Fill the plastic burette with 25 mL of 0.1 M NaOH. Drain about 5 mL into the waste beaker&#8212;enough so that NaOH fills the burette tip. Make sure there are no air bubbles, then close the valves.</div>
		</li>
		<li data-sub-note-item="3-6">
		<div class="lab-step">Calibrate the drop counter. Replace the waste beaker beneath the burette with a 10-mL graduated cylinder. Then, open the bottom valve on the burette, while keeping the top valve closed. Turn on the data acquisition system and set it to &#8216;drop counting mode&#8217;.</div>
		</li>
		<li data-sub-note-item="3-7">
		<div class="lab-step">Slowly open the top valve to very slowly release drops, ideally at one drop every 2 s. Allow the drops to empty from the burette until there is 9-10 mL of 0.1 M NaOH in the graduated cylinder.</div>
		</li>
		<li data-sub-note-item="3-8">
		<div class="lab-step">Close the bottom valve and leave the top valve as is. Read the volume of NaOH in the graduated cylinder to the first decimal place and enter this value in the data acquisition system. Record the value for &#8216;drops/mL&#8217;. Then, discard the NaOH in the graduated cylinder into the aqueous waste beaker.</div>
		</li>
		<li data-sub-note-item="3-9">
		<div class="lab-step">Calibrate the pH sensor before starting the titration. Connect the pH sensor to the data acquisition system, then select &#39;Calibrate&#39;. Rinse the bulb of the pH sensor with deionized water before inserting it into the pH 7 buffer. Leave the sensor submerged until the voltage stabilizes, then accept the measurement.</div>
		</li>
		<li data-sub-note-item="3-10">
		<div class="lab-step">Rinse the bulb with deionized water and insert it into the pH 10 buffer. Allow the voltage to stabilize, then accept the measurement.</div>
		</li>
		<li data-sub-note-item="3-11">
		<div class="lab-step">Rinse the bulb of the pH sensor again and slide it through the designated slot in the drop counter.</div>
		</li>
		<li data-sub-note-item="3-12">
		<div class="lab-step">Measure 40 mL of deionized water into a clean 100-mL beaker, then, transfer 1 mL of phosphoric acid into the beaker of water.</div>
		</li>
		<li data-sub-note-item="3-13">
		<div class="lab-step">Place the beaker on the stir plate under the drop counter. Carefully slide the pH sensor into the beaker.</div>
		</li>
		<li data-sub-note-item="3-14">
		<div class="lab-step">Add a stir bar to the beaker, and turn the stir setting onto high. Start collecting data on the acquisition device. Then, open the bottom valve on the burette. <strong>Note: </strong>The drop rate should be about 1 drop every 2 s. After the first drop is released, check to see data are being recorded.</div>
		</li>
		<li data-sub-note-item="3-15">
		<div class="lab-step">Continue the titration until the pH meter reads pH 12. Then, stop acquiring data and close the valve on the burette. Save your data on a flash drive.</div>
		</li>
		<li data-sub-note-item="3-16">
		<div class="lab-step">To clean up your workspace, check the pH of all waste solutions using pH paper. Neutralize all acidic aqueous waste with baking soda and all basic waste with citric acid. Add enough of either baking soda or citric acid to the solution until it stops bubbling.</div>
		</li>
		<li data-sub-note-item="3-17">
		<div class="lab-step">Flush all neutralized solutions down the sink with copious amounts of water.</div>
		</li>
		<li data-sub-note-item="3-18">
		<div class="lab-step">Wash all glassware.</div>
		</li>
	</ul>
	</li>
	<li data-note-item="4" data-parent-collapse="">Results<button class="expand">Expand</button>
	<ul class="lab-items">
		<li data-sub-note-item="4-1">
		<div class="lab-step">In the first part of this lab, you standardized a solution of NaOH using KHP to determine its actual concentration. Now, let&#39;s see how close the standardized concentration is to the 0.1 M concentration that was prepared. Determine the number of moles of KHP that was added to each flask, and by extension, the moles of NaOH. Once the solution is neutralized, the molar quantities of KHP and NaOH are equal.</div>
		</li>
		<li data-sub-note-item="4-2">
		<div class="lab-step">Calculate the molarity of NaOH based on the total volume of NaOH that was added to each flask. The actual concentration is lower than the expected 0.1 molarity. This is because NaOH is hygroscopic, so it is difficult to weigh it accurately.</div>
		</li>
		<li data-sub-note-item="4-3">
		<div class="lab-step">Plot the results for the phosphoric acid titration (pH vs. Volume of NaOH). Phosphoric acid is a weak triprotic acid, meaning that it has the potential to provide three protons per molecule when it dissociates in aqueous solutions. Phosphoric acid has three pKa values, one for when each proton is dissociated.</div>
		</li>
		<li data-sub-note-item="4-4">
		<div class="lab-step">Look at the data. There are two sigmoidal curves, indicating two equivalence points. Each equivalence point corresponds to a dissociation constant, Ka, of phosphoric acid. <strong>Note: </strong>You stopped the experiment once the pH reached 12, so you only measured two of the three Ka values.</div>
		</li>
		<li data-sub-note-item="4-5">
		<div class="lab-step">Plot the first derivative of the titration curve. The equivalence points are represented by the curve maximums.
		<h4><strong>Table 3. Titration of phosphoric acid</strong></h4>
		<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
			<tbody>
				<tr>
					<td>Volume<sub>1<sup>st</sup> equivalence point</sub> (mL)</td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>Volume<sub>1<sup>st</sup> half-equivalence point</sub> (mL)</td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>Volume<sub>2<sup>nd</sup> equivalence point</sub> (mL)</td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>Volume<sub>2<sup>nd</sup> half-equivalence point</sub> (mL)</td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>1<sup>st</sup> pKa<sub>measured</sub></td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>1<sup>st</sup> pKa<sub>theoretical</sub></td>
					<td style="text-align: center;">2.16</td>
				</tr>
				<tr>
					<td>2<sup>nd</sup> pKa<sub>measured</sub></td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>2<sup>nd</sup> pKa<sub>theoretical</sub></td>
					<td style="text-align: center;">7.21</td>
				</tr>
				<tr>
					<td>Moles of NaOH</td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>Moles of H<sub>3</sub>PO<sub>4</sub></td>
					<td style="text-align: center;"></td>
				</tr>
				<tr>
					<td>Molarity of H<sub>3</sub>PO<sub>4</sub></td>
					<td style="text-align: center;"></td>
				</tr>
			</tbody>
		</table>
		<a href="https://www-jove-com.remotexs.ntu.edu.sg/CDNSource/lm/labs/44/Table_3._Titration_of_Phosphoric_Acid.xlsx" target="_blank">Click Here to download Table 3</a></div>
		</li>
		<li data-sub-note-item="4-6">
		<div class="lab-step">Find the first half-equivalence point by taking the volume of NaOH corresponding to the first equivalence point and dividing it by 2. At the half-equivalence point, the concentrations of undissociated acid and its conjugate base are the same, and the pH is equal to the pKa.</div>
		</li>
		<li data-sub-note-item="4-7">
		<div class="lab-step">Look up the pH at this volume from your table of data to get a more precise value. This corresponds to the first pKa, which is reported in the literature as 2.16.</div>
		</li>
		<li data-sub-note-item="4-8">
		<div class="lab-step">Repeat this to find the second pKa. The second half-equivalence point is located midway between the first and second equivalence points, which should give a pKa of about 7.2.</div>
		</li>
	</ul>
	</li>
</ol>

Acid-Base Titration: Calculating pH, Strength, and Concentration - Procedure

Source: Smaa Koraym at Johns Hopkins University, MD, USA

	Preparation of ~0.1 M NaOHExpand
	In the first part of the lab, you will use a 50% w/w solution ...

Enthalpy of Reaction

Enthalpy of Reaction and Measurement of Heat by Calorimeter - procedure

Source: Smaa Koraym at Johns Hopkins University, MD, USA

	Collecting Data for Calculating the Heat Capacity of a CalorimeterExpand
	This experiment will ...

Physiology of the Circulatory System

<ol class="note-items"><li data-note-item="1" data-parent-collapse="">Measuring Circulatory System Function in Humans
<button class="expand">Expand</button><ul class="lab-items"><li data-sub-note-item="1-1"><div class="lab-step">Before starting the experiment go to the front of the room and collect an alcohol swab, a sphygmomanometer, and a stethoscope.
</div></li><li data-sub-note-item="1-2"><div class="lab-step">Clean the earpieces with the alcohol swab and then insert them into your ears.
</div></li><li data-sub-note-item="1-3"><div class="lab-step">To check if the stethoscope is in the on position, gently tap the flat metal disc called the diaphragm.
</div></li><li data-sub-note-item="1-4"><div class="lab-step">If you hear a sound it&#39;s on. If not, turn the metal disc and tap again and keep turning and tapping the disc until you hear a sound. NOTE: For this experiment you will work in pairs. One person will measure their partner&#39;s blood pressure, while the person being measured will count their heartbeats over the course of one minute.
</div></li><li data-sub-note-item="1-5"><div class="lab-step">Fill out table one with the name of the person being monitored. HYPOTHESES: The experimental hypothesis is that systolic blood pressure will be higher when the person is sitting when compared to lying down because more pressure is required for blood to be moved around the body. The null hypothesis is that there will be no difference in systolic blood pressure whether a person is sitting or lying down.<a href="https://www-jove-com.remotexs.ntu.edu.sg/CDNSource/lm/labs/9/Table_1_Physiology_of_Circulatory_System.xlsx" style="display:block;margin:20px;" target="__blank">Click Here to download Table 1</a>
</div></li><li data-sub-note-item="1-6"><div class="lab-step">The person being monitored should begin by lying down. Next, they will need to locate their pulse by placing two fingers next to their windpipe. If a throbbing sensation is felt, this is the pulse.
</div></li><li data-sub-note-item="1-7"><div class="lab-step">Now, ready a time keeping device to count out 1 minute as the person taking their pulse will be having their blood pressure taken at the same time.
</div></li><li data-sub-note-item="1-8"><div class="lab-step">At this point, the person measuring blood pressure should place the cuff of the sphygmomanometer around the other arm of the subject and the stethoscope earpieces into their own ears.
</div></li><li data-sub-note-item="1-9"><div class="lab-step">Start the timer.
</div></li><li data-sub-note-item="1-10"><div class="lab-step">The person being monitored should begin counting the number of heartbeats felt over one full minute. While the subject is taking their pulse, the person taking blood pressure should place the diaphragm of the stethoscope under the cuff on the inside of their partner&#39;s elbow.
</div></li><li data-sub-note-item="1-11"><div class="lab-step">Then, pump the cuff to 200 mm of mercury.
</div></li><li data-sub-note-item="1-12"><div class="lab-step">Gently and slowly twist the metal knob on the sphygmomanometer to release air from the cuff.
</div></li><li data-sub-note-item="1-13"><div class="lab-step">As pressure is released the first Korotkoff sound should be heard. The corresponding value on the sphygmomanometer is the systolic blood pressure. Consistent sounds should be heard until they die out with the last sound signifying the diastolic blood pressure.
</div></li><li data-sub-note-item="1-14"><div class="lab-step">Record the systolic pressure, the diastolic pressure, and the heart rate in Table 1.
</div></li><li data-sub-note-item="1-15"><div class="lab-step">Now, repeat the blood pressure and pulse measurements with each partner keeping the same role but this time the subject should be sitting upright.
</div></li><li data-sub-note-item="1-16"><div class="lab-step">Record these data in the appropriate places in Table 1.
</div></li><li data-sub-note-item="1-17"><div class="lab-step">Finally, the partners should switch spots. So now, the person measuring blood pressure should be taking their pulse and acting as the subject.  NOTE: Be sure to wipe the earpieces of the stethoscope between users and don&#39;t forget to record your measurements in Table 1.
</div></li><li data-sub-note-item="1-18"><div class="lab-step">After finishing the experiment, discard all alcohol swabs into the trash. Then, bring the sphygmomanometer and stethoscope back to the front of the room.
</div></li></ul></li><li data-note-item="2" data-parent-collapse="">Measuring Respiratory Rate in Fish
<button class="expand">Expand</button><ul class="lab-items"><li data-sub-note-item="2-1"><div class="lab-step">To prepare for this second experiment, collect two clear containers.
</div></li><li data-sub-note-item="2-2"><div class="lab-step">Begin by filling half of one container with aquarium water.
</div></li><li data-sub-note-item="2-3"><div class="lab-step">Then, use a fish net to gently catch a goldfish and place it into the cup. Pinch the net around the fish or else it may jump out of the net.
</div></li><li data-sub-note-item="2-4"><div class="lab-step">Familiarize yourself with the operculum of the fish. The operculum covers the gills and it is located behind and below the eye. When the fish respires the operculum moves in and out and this is one gill beat. HYPOTHESES: In this experiment you will measure the gill beats of the fish in different water temperatures. The hypothesis is that there will be a significant correlation between the temperature and respiration rate in goldfish. The null hypothesis is that there will be no correlation between temperature and respiration rate.
</div></li><li data-sub-note-item="2-5"><div class="lab-step">Take the other empty container with a screw cap, the cup with your fish, and the net to the water tanks.
</div></li><li data-sub-note-item="2-6"><div class="lab-step">Then, fill the empty container with the water from the lowest temperature water bath. This will act as your first respiration rate chamber.
</div></li><li data-sub-note-item="2-7"><div class="lab-step">Next, hold the fish net over a wastewater container and dump out the water from the cup holding the fish so that the fish falls into the net. Pinch the net around the fish without touching the fish itself.
</div></li><li data-sub-note-item="2-8"><div class="lab-step">Bring the net over the top of the container with the bath water and let go of the net to release the fish into the vial.
</div></li><li data-sub-note-item="2-9"><div class="lab-step">Once the fish is in the container, submerge it back into the water bath and then screw the cap on to prevent outside oxygen from entering the vial.
</div></li><li data-sub-note-item="2-10"><div class="lab-step">Next, remove the container from the controlled temperature water bath and place it on your workbench surface to avoid warming the water with body heat.
</div></li><li data-sub-note-item="2-11"><div class="lab-step">One partner should immediately begin counting the gill beats as soon as the second partner starts the stopwatch.
</div></li><li data-sub-note-item="2-12"><div class="lab-step">Continue counting the beats for 20 seconds. Record the number of beats in column A of Table 2. <a href="https://www-jove-com.remotexs.ntu.edu.sg/CDNSource/lm/labs/9/Table_2_Physiology_of_Circulatory_System.xlsx" style="display:block;margin:20px;" target="__blank">Click Here to download Table 2</a>
</div></li><li data-sub-note-item="2-13"><div class="lab-step">Repeat the measurement one more time and record the number in column B.
</div></li><li data-sub-note-item="2-14"><div class="lab-step">Next, hold the fish net over a wastewater container and dump out the water from the fish vial so that the fish falls into the net.  IMPORTANT: Move from lowest to highest temperature, so the fish is not stressed between trials.
</div></li><li data-sub-note-item="2-15"><div class="lab-step">Repeat the experiment for the remaining four temperature conditions (steps 8 &#8211; 13).
</div></li><li data-sub-note-item="2-16"><div class="lab-step">Record all the values for different temperatures in Table 2.
</div></li><li data-sub-note-item="2-17"><div class="lab-step">Once you are done, slowly mix water from the last respiration chamber with the fish tank water and then return the fish to the home tank. NOTE: This should be done over the course of approximately 10 minutes.
</div></li></ul></li><li data-note-item="3" data-parent-collapse="">Results
<button class="expand">Expand</button><ul class="lab-items"><li data-sub-note-item="3-1"><div class="lab-step">It&#39;s time to analyze the data that you just collected. Look at the blood pressure and heart rate measurements for the different trials. If they were different, make a hypothesis to explain your observations. 
</div></li><li data-sub-note-item="3-2"><div class="lab-step">For the gill beat experiment, average the numbers in columns A and B and calculate the fish respiration rate as gill beats per minute.
</div></li><li data-sub-note-item="3-3"><div class="lab-step">Next, plot the graph of temperature versus gill beats per minute.
</div></li><li data-sub-note-item="3-4"><div class="lab-step">Record any correlations you see between the variables. If a correlation is observed, determine if it is positive or negative and provide an explanation for this result.  NOTE: Consider the fact that fishes are ectotherms and the impact of temperature on dissolved oxygen.
</div></li></ul></li></ol>

Measuring Circulatory System Function in Humans
ExpandBefore starting the experiment go to the front of the room and collect an alcohol swab, a ...

Biology

Fundamentals

Cellular Respiration

<ol class="note-items"><li data-note-item="1" data-parent-collapse="">Quantifying Respiration using Microrespirometers
<button class="expand">Expand</button><ul class="lab-items"><li data-sub-note-item="1-1"><div class="lab-step">To assemble the microrespirometers needed for the lab, first plug in the hot glue gun to preheat it.
</div></li><li data-sub-note-item="1-2"><div class="lab-step">Push the plunger of a tuberculin syringe all the way in and then cut the end off of the syringe.
</div></li><li data-sub-note-item="1-3"><div class="lab-step">Next, insert a capillary tube into the empty needle end of the syringe.
</div></li><li data-sub-note-item="1-4"><div class="lab-step">Push the capillary tube in until it touches the tip of the plunger.
</div></li><li data-sub-note-item="1-5"><div class="lab-step">Use the hot glue gun to create an airtight seal between the capillary tube and the tuberculin syringe. Make sure that the capillary tube is standing straight up while the glue dries.
</div></li><li data-sub-note-item="1-6"><div class="lab-step">Once the glue has dried, pull back on the plunger and make sure the capillary tube is not plugged.
</div></li><li data-sub-note-item="1-7"><div class="lab-step">Next, prepare the manometer fluid by adding red food coloring to soapy water. NOTE: The detergent coats the inside of the capillary tube, preventing the manometer fluid from sticking during the experiment.
</div></li><li data-sub-note-item="1-8"><div class="lab-step">Insert the capillary tube into the manometer fluid and then pull the plunger on the syringe until the entire capillary tube has been filled.
</div></li><li data-sub-note-item="1-9"><div class="lab-step">Then, eject the manometer fluid back out of the microrespirometer.
</div></li><li data-sub-note-item="1-10"><div class="lab-step">With a small piece of absorbent cotton, plug the barrel of the microrespirometer, inserting it into the syringe up to the 0 mL mark. Pack the cotton using a spare capillary tube or a glass disposable pipette.
</div></li><li data-sub-note-item="1-11"><div class="lab-step">Add a single drop of 15% KOH to the cotton using a spare capillary tube or a glass disposable pipette.
</div></li><li data-sub-note-item="1-12"><div class="lab-step">Add some nonabsorbent cotton to the KOH saturated cotton to act as a barrier to the seeds. Pack the cotton using a disposable pipette. The cotton should reach halfway between the 0 mL and 1 mL mark.
</div></li><li data-sub-note-item="1-13"><div class="lab-step">Reinsert the plunger carefully until it reaches the cotton and then remove any excess KOH by pressing it out.
</div></li><li data-sub-note-item="1-14"><div class="lab-step">Set four microrespirometers at each lab station.
</div></li><li data-sub-note-item="1-15"><div class="lab-step">Then, place a lab marker and two small dishes containing the seeds and glass beads for each group alongside the microrespirometers.
</div></li><li data-sub-note-item="1-16"><div class="lab-step">Finally, prepare two water baths by filling them with roughly 6 inches of water. The water level in the bath should comfortably fit two respirometers with the entirety of their two capillary tubes exposed.
</div></li><li data-sub-note-item="1-17"><div class="lab-step">Replace the lids and insert a thermometer in each bath to monitor the temperature. One bath should be set at about 25 &#176;C, and the other at approximately 40 &#176;C.
</div></li></ul></li></ol>

Cellular Respiration: Quantification using Microrespirometers - Prep

Quantifying Respiration using Microrespirometers
ExpandTo assemble the microrespirometers needed for the lab, first plug in the hot glue gun to preheat ...