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Absorbance Spectroscopy | Absorbance Explained | Ossila

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This can provide important information about a samples molecular structure, concentration or optoelectronic properties. Depending on the sample, absorban" /><link rel="apple-touch-icon" sizes="180x180" href="//www.ossila.com/cdn/shop/t/303/assets/apple-touch-icon.png?v=47689890648139992551729065668"> <link rel="icon" type="image/png" sizes="32x32" href="//www.ossila.com/cdn/shop/t/303/assets/favicon-32x32.png?v=170914528706734454101729065668"> <link rel="icon" type="image/png" sizes="16x16" href="//www.ossila.com/cdn/shop/t/303/assets/favicon-16x16.png?v=162550173345868048431729065668"> <link rel="manifest" href="//www.ossila.com/cdn/shop/t/303/assets/site.webmanifest?v=100969836723433583841729065669"> <link rel="mask-icon" href="//www.ossila.com/cdn/shop/t/303/assets/safari-pinned-tab.svg?v=95647162979764929801729065669" color="#39499c"> <meta name="msapplication-TileColor" content="#418fde"> <meta name="theme-color" content="#ffffff"> <link rel="canonical" href="https://www.ossila.com/pages/absorbance-spectroscopy" /> <!-- CSS --> <link rel="stylesheet" href="//www.ossila.com/cdn/shop/t/303/assets/main-style.css?v=129786468726129743101731684242"> <!-- JS --> <script>var shopifyMoneyFormat = '${{amount}}';</script> <script defer src="//www.ossila.com/cdn/shop/t/303/assets/main.js?v=120109391065215852831729088283"></script> <!--[template]-[page]--> <meta property="og:type" content="website" /> <meta property="og:title" content="Absorbance Spectroscopy | Absorbance Explained" /> <meta property="og:description" content="In absorbance spectroscopy (also known as absorption spectroscopy), an USB spectrometer measures the amount of light absorbed by a sample as a function of wavelength. This can provide important information about a samples molecular structure, concentration or optoelectronic properties. Depending on the sample, absorban" /> <meta property="og:url" content="https://www.ossila.com/pages/absorbance-spectroscopy" /> <meta property="og:site_name" content="Ossila" /> <meta name="twitter:site" content="@ossilascience"> <meta name="twitter:card" content="summary"> <meta name="twitter:title" content="Absorbance Spectroscopy"> <meta name="twitter:description" content="In absorbance spectroscopy (also known as absorption spectroscopy), an USB spectrometer measures the amount of light absorbed by a sample as a function of wavelength. This can provide important information about a samples molecular structure, concentration or optoelectronic properties. Depending on the sample, absorbance measurements can also give you key insights into dynamic material properties, such as phase or composition changes. To measure absorbance, calculate the logarithmic of the ratio of the initial light intensity and light intensity transmitted through the sample. Absorbance values are unitless and always wavelength dependent. However, for solution and solid samples other values can be used, such as the molar attenuation coefficient and extinction coefficient respectively. Contents Absorbance Theory Defining Absorbance Molar Attenuation Coefficient Absorption Coefficient Absorbance Units Absorbance Uses Resources and References Absorbance Theory Within an atom, electrons exist in regions around the nucleus known as orbitals. 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You can still place orders by emailing us on <a href="mailto:info@ossila.com">info@ossila.com</a>, but you may experience issues browsing our website. Please consider upgrading to a modern browser for better security and an improved browsing experience.</p> </div> </div> <!-- no sidebars --> <main> <div id="informational-page"> <h1>Absorbance Spectroscopy</h1> <div class="container"> <img alt="Absorbance Spectroscopy" class="featured-image" height="280" loading="lazy" src="https://www.ossila.com/cdn/shop/files/Ossila-Optical-Spectrometer-Kit-Fluorescence.jpg?crop=center&amp;width=1340&amp;height=280" width="1340"> <figure class="double-float-right margin-top"><video alt="Absorbance Spectroscopy electron excitation" autoplay="autoplay" loop="loop" muted="muted" src="https://cdn.ossila.com/videos/electron-excitation.mp4" type="video/mp4" title="Absorbance Spectroscopy electron excitation"></video></figure> <p>In absorbance spectroscopy (also known as absorption spectroscopy), an <a href="/products/optical-spectrometer" title="USB Spectrometer">USB spectrometer</a> measures the amount of light absorbed by a sample as a function of wavelength. This can provide important information about a samples molecular structure, concentration or optoelectronic properties. Depending on the sample, absorbance measurements can also give you key insights into dynamic material properties, such as phase or composition changes.</p> <p>To measure absorbance, calculate the logarithmic of the ratio of the initial light intensity and light intensity transmitted through the sample. Absorbance values are unitless and always wavelength dependent. However, for solution and solid samples other values can be used, such as the molar attenuation coefficient and extinction coefficient respectively.</p> <br> <br> <br> <br> <br> <div class="panel panel-default"> <div class="panel-body"> <p class="h3" style="margin-top: 10px;">Contents</p> <hr> <ul> <li><a href="#absorbance-theory">Absorbance Theory</a></li> <li><a href="#defining-absorbance">Defining Absorbance</a></li> <ul> <li><a href="#molar-attenuation-coefficient">Molar Attenuation Coefficient</a></li> <li><a href="#absorption-coefficient">Absorption Coefficient</a></li> </ul> <li><a href="#absorbance-units">Absorbance Units</a></li> <li><a href="#absorbance-uses">Absorbance Uses</a></li> <li><a href="#resources-references">Resources and References</a></li> </ul> </div> </div> <h2 id="absorbance-theory">Absorbance Theory</h2> <hr> <p>Within an atom, electrons exist in regions around the nucleus known as orbitals. Depending on the electronic structure of an atom, these orbitals (or energy levels) are either completely filled, partially filled, or empty. When you bring multiple atoms together in a molecule, solid or solution, this will change its orbitals, electron distribution and therefore the samples electronic and optical properties.</p> <p>Absorbance simply tells you the amount of light that is absorbed by a sample. However, by measuring the wavelengths of light that are absorbed by a material, you can probe these different energy levels and study how electrons move between them - which can give you some useful information about your sample.</p> <p>For many materials, absorption occurs when the the energy of an incoming photon (E<sub>photon</sub>) is equal or larger than energy difference (ΔE) between the highest occupied molecular orbit (HOMO) and the lowest unoccupied molecular orbit (LUMO).</p> <figure><img loading="lazy" alt="In absorbance spectroscopy, the photon energy must be larger than the band gap" height="17" src="https://www.ossila.com/cdn/shop/files/equation-energy-photon-homo-lumo-absorption.png?v=1725544897" width="207"></figure> <p>If its energy is high enough, an incoming photon can be absorbed by an electron in the HOMO level. It is then be excited into a higher energy state. This light will not be transmitted through the sample, hence it had been absorbed.</p> <figure><img alt="Photon absorbance in a molecule with energy gap, ΔE" height="305.73" loading="lazy" src="https://www.ossila.com/cdn/shop/files/Absorbance_Spectroscopy.svg?v=1677861116" width="400"> <figcaption>Photon absorbance in a molecule with energy gap, <em>ΔE</em>. If the photon is <em>E<sub>ph</sub> &gt; ΔE</em>, an electron will absorb the photon, and it can be excited from the HOMO to the LUMO. However, if the photon is less than this energy, it will not be absorbed.</figcaption> </figure> <p>Some materials however can only absorb light at very specific energies. This is often described in terms of the photons wavelength (λ). Wavelengths of the absorbed light correspond to energy through the following equation.</p> <figure><img alt="Equation - energy of a photon is equal to plancks constant times the speed of light, all divided by the photons wavelength" height="40.953" loading="lazy" src="https://www.ossila.com/cdn/shop/files/equation-photon-energy.svg?v=1718880486" width="63"></figure> <p>The types of electronic transitions that are allowed often vary with different material properties. Measuring the wavelengths at which absorbance occurs will therefore tell you a lot about the electronic properties of a material, molecule or thin film.</p> <p>Measuring absorbance requires a broadband light source and an <a href="/products/optical-spectrometer" title="USB Spectrometer">USB spectrometer</a> with appropriate spectral range. Absorbance values are closely related to transmission measurements, as in both cases you actually measure the transmitted light.</p> <figure><img alt="Equipment needed for absorbance measurements" height="371.141" loading="lazy" src="https://www.ossila.com/cdn/shop/files/Optical-Spectroscopy-Transmittance.svg?v=1676996706" width="848.328"> <figcaption>Light is transmitted through the sample and recorded by the spectrometer</figcaption> </figure> <p>As light moves through a sample, it is more likely to be absorbed. Therefore, the intensity of transmitted light will fall exponentially with distance. You can calculate absorbance by the comparing the reference light spectrum, I<sub>0,</sub> to the detected light that has passed through the sample, I, using this equation.</p> <figure><img alt="Absorbance Equation - Light Intensity" height="47.8" loading="lazy" src="https://www.ossila.com/cdn/shop/files/equation-absorbance.svg" width="129.55"></figure> <h2 id="defining-absorbance">Defining Absorbance</h2> <hr> <p>Absorbance is a relative measurement so is therefore unitless. Having said this, absorption properties of different samples can be defined in a number of different ways.</p> <h3 id="molar-attenuation-coefficient"><strong>Molar Attenuation Coefficient</strong></h3> <p>When you measure the absorbance spectrum of a solution, one important factor is the wavelengths where maximum absorbance occurs. Identifying these wavelengths can can help you determine certain molecular properties of your sample. You can also note the strength of absorbance at these wavelengths.</p> <p>By looking at these peaks, you can establish the molar attenuation coefficient, ε. This has also been referred to historically as the molar extinction coefficient and is also known as the molar absorptivity coefficient. This will tell you about the kinds of electronic transitions that are occurring within your sample associated. For allowed transitions, ε &gt; 1000 whereas for forbidden transitions, ε &lt; 100.</p> <p>The molar attenuation coefficient, ε, is featured in Beer Lambert's Law:</p> <figure><img alt="Beer Lamberts Law Equation" height="" loading="lazy" src="https://www.ossila.com/cdn/shop/files/equation-beer-lambert-law-absorbance.svg?v=1719214248" width="120"></figure> <p>Where A is absorbance, c is the molar concentration of the molecule in solution, and l is the path length through the sample (often the width of the cuvette, or the total film). You can use this calculation to measure the concentration of a molecule in a thin film. The graphs below show the variation in absorbance intensity with concentration.</p> <figure><img alt="Absorption spectrum of a film made from a solution of polystyrene in toluene with varying concentrations of BODIPY-Br at varying concentrations" height="306.09" loading="lazy" src="https://www.ossila.com/cdn/shop/files/BODIPY_OD.png?v=1674130763&amp;width=400&amp;height=306" width="400"><img alt="Linear dependence of absorbance at λmax (531 - 533 nm) compared to BOPIDY concentration" height="230.08" loading="lazy" src="https://www.ossila.com/cdn/shop/files/Absorbance-concentration-BOPIDY.png?v=1674130874&amp;width=400&amp;height=230" width="400"> <figcaption>Left.) Absorption spectrum of a film made from a solution of polystyrene in toluene with varying concentrations of BODIPY-Br at varying concentrations. Right.) Linear dependence of absorbance at λ<sub>max</sub> (531 - 533 nm) compared to BOPIDY concentration.</figcaption> </figure> <p>Molar attenuation coefficient has property has dimensions of</p> <figure><img alt="Molar attenuation coefficient equation" height="59.75" loading="lazy" src="https://www.ossila.com/cdn/shop/files/equation-molar-absorbtivity-units.svg?v=1674130982" width="490.92"></figure> <p>Therefore, the SI units of the molar absorptivity coefficient is m<sup>2</sup>M<sup>-1 </sup>or cm<sup>2</sup>M<sup>-1</sup>.</p> <h3 id="absorption-coefficient"><strong>Absorption Coefficient</strong></h3> <p>For bulk solids, you can define absorbance in terms of an absorption coefficient, α(λ). This is a property of a solid material and is wavelength dependent. The absorption coefficient, α(λ) relates to absorbance measurements through the following equation:</p> <figure><img alt="Absorption Coefficient equation" height="19.94" loading="lazy" src="https://www.ossila.com/cdn/shop/files/equation-absorption-coefficient.svg?v=1674131050" width="105.22"></figure> <p>Here, d is the distance light has travelled into (or through) the bulk solid.</p> <p>The SI units of the absorption coefficient of a bulk solid are m<sup>-1</sup> or cm<sup>-1</sup>.</p> <h2 id="absorbance-units">Absorbance Units</h2> <hr> <table> <thead> <tr> <th width="25%">Property</th> <th width="37.5%">Units</th> <th width="37.5%">Sample Type</th> </tr> </thead> <tbody> <tr> <td>Absorbance</td> <td>Unitless</td> <td>General</td> </tr> <tr> <td>Absorption Coefficient</td> <td>m<sup>-1</sup> </td> <td>Bulk Solid</td> </tr> <tr> <td>Molar Absorptivity Coefficient</td> <td>m<sup>2</sup>M<sup>-1</sup> </td> <td>Molecule in solution</td> </tr> </tbody> </table> <h2 id="absorbance-uses">Absorbance Uses</h2> <hr> <p>You can use absorbance spectroscopy to investigate the suitability of a material for specific purposes. For example:</p> <ul> <li>Organic dyes require high absorbance over a small wavelength range so that only light of a matching colour is emitted from the dye, e.g. for a yellow dye, blue light (435 - 480 nm) must be absorbed.</li> <li>Materials that convert visible light into different forms of energy (such as solar cells, photosensors) will require high absorption, ideally across the visible region. These materials frequently have desirable band gaps, which can be determined using absorbance spectroscopy.</li> <li>Transparent materials need to absorb as little visible light as possible. You can confirm this using UV-Vis spectroscopy.</li> <li>In organic compounds, UV-vis spectroscopy can help illuminate the amount of conjugated pi bonds in a molecule.</li> </ul> <a class="green-banner" data-track-content="" href="https://www.ossila.com/products/optical-spectrometer"> <p data-content-name="/pages/absorbance-spectroscopy" data-content-piece="cta-optical-spectrometer">USB Spectrometer</p> <img alt="USB Spectrometer" height="120" loading="lazy" src="https://www.ossila.com/cdn/shop/files/Ossila-Optical-Spectrometer-Kit-Fluorescence.jpg?crop=center&amp;width=600&amp;height=120" width="600"> <button>Browse now</button> </a> <h2 id="resources-references">Resources</h2> <hr> <div class="tile-container"> <div class="card-box card-truncate"> <a href="/pages/absorbance-measurements"> <img alt="Glove Box electrical feedthrough assembly" height="160" loading="lazy" src="https://www.ossila.com/cdn/shop/files/optical-spectroscopy-example-spectra-absorbance.png?v=1657537655" width="160"> </a> <a href="/pages/absorbance-measurements"> Absorbance Measurement</a> <p>Absorbance measurements are crucial in many areas of scientific research. This article describes how to take an absorbance measurement using an optical spectrometer.</p> <a href="/pages/absorbance-measurements">Read more...</a> </div> <div class="card-box card-truncate"> <a href="/pages/spectrometer-buying-guide"> <img alt="Choosing a spectrometer" height="160" loading="lazy" src="https://www.ossila.com/cdn/shop/files/ossila-spectrometer-in-lab.jpg?v=1631265957&amp;crop=center&amp;width=160&amp;height=160" width="160"></a> <a href="/pages/spectrometer-buying-guide">Choosing a Spectrometer</a> <p>When researching optical materials, an optical spectrometer is an essential instrument which enables you to characterize your materials quickly and easily. It is a powerful tool that can be used to measure the properties of light such as wavelength and intensity</p> <a href="/pages/spectrometer-buying-guide">Read more...</a> </div> </div> <div class="panel panel-default"> <div class="panel-body"> <h3>Contributing Authors</h3> <hr> <p>Written by</p> <p class="no-margin"><a href="/pages/our-writers#mary-okane" title="Author Biography: Mary O'Kane">Dr. Mary O'Kane</a></p> <p class="text-muted">Application Scientist</p> </div> </div> <div class="panel panel-default"> <div class="panel-body"> <h3>References</h3> <hr> <ol> <li>Rocha, F. S., Gomes, A. J., Lunardi, C. N., Kaliaguine, S., &amp; Patience, G. S. (2018). Experimental methods in chemical engineering: Ultraviolet visible spectroscopy-UV-vis. <i>The Canadian Journal of Chemical Engineering</i>, <i>96</i>(12), 2512–2517. DOI:10.1002/cjce.23344</li> <li>Hestand, N. J., &amp; Spano, F. C. (2018). Expanded theory of H- and J-molecular aggregates: The effects of vibronic coupling and intermolecular charge transfer. <i>Chemical Reviews</i>, <i>118</i>(15), 7069–7163. DOI: 10.1021/acs.chemrev.7b00581</li> <li>Makuła, P., Pacia, M., &amp; Macyk, W. (2018). How to correctly determine the band gap energy of modified semiconductor photocatalysts based on UV–vis spectra. <i>The Journal of Physical Chemistry Letters</i>, <i>9</i>(23), 6814–6817. DOI: 10.1021/acs.jpclett.8b02892</li> <li>Chen, J., Zhou, S., Jin, S., Li, H., &amp; Zhai, T. (2016). Crystal organometal halide perovskites with promising optoelectronic applications. <i>Journal of Materials Chemistry C</i>, <i>4</i>(1), 11–27. DOI: 10.1039/c5tc03417e</li> </ol> </div> </div> <script type="application/ld+json"> { "@context": "https://schema.org", "@type": "Article", "abstract": "In absorbance spectroscopy (also known as absorption spectroscopy), a spectrometer is used to directly measure how much light is absorbed by a sample as a function of wavelength. Quantifying the absorbance of light by an atom or molecule can provide important information about the molecules electronic structure. This guide explains the absorbance theory and how to measure it as well as the uses for absorbance.", "articleSection": ["Optical Spectroscopy"], "author": [ { "@type": "Person", "name": "Mary O'Kane", "honorificPrefix": "Dr", "jobTitle": "Application Scientist", "url": "https://www.ossila.com/pages/our-writers#mary-okane" } ], "headline": "Absorbance Spectroscopy", "image": [ "https://www.ossila.com/cdn/shop/files/Optical-Spectroscopy-Transmittance.svg?v=1676996706" ], "inLanguage": "en", "publisher": { "@type": "Organization", "name": "Ossila", "url": "https://www.ossila.com" }, "url": "https://www.ossila.com/pages/absorbance-spectroscopy" } </script> </div> </div> </main> <!-- Begin Footer --> <div class="full-width-footer"> <footer id="footer" class="footer container"> <div class="clearfix footer-elems nav-footer" id="footer-links"> <div class="row"> <div class="col-xs-12 col-sm-3"> <span class="h2">Ossila</span> <ul class="nav-footer"> <li><a href="/pages/about">About Ossila</a></li> <li><a href="/pages/press-and-media">Press and Media</a></li> <li><a href="/pages/our-writers">Our Writing Team</a></li> <li><a href="/pages/awards-and-testimonials">Awards and Testimonials</a></li> <li><a href="/pages/feedback">Leave Feedback</a></li> <li><a href="/pages/terms-and-conditions">Terms and Conditions</a></li> <li><a href="/pages/privacy-policy">Privacy Policy</a></li> </ul> <span>Copyright &copy; 2024 Ossila</span> </div> <div class="col-xs-12 col-sm-3"> <span class="h2">Join the Team</span> <ul class="nav-footer"> <li><a href="/pages/careers">Careers at Ossila</a></li> <li><a href="/pages/jobs-in-sheffield">Jobs in Sheffield</a></li> <li><a href="/pages/jobs-in-leiden">Jobs in Leiden</a></li> </ul> <span class="h2">Collaborate</span> <ul class="nav-footer"> <li><a href="/pages/reprinting-permissions">Free Digital Assets</a></li> <li><a href="/pages/conference-fund">Conference Sponsorship for Students</a></li> <li><a href="/pages/outreach-support">Science Outreach Support</a></li> <li><a href="/pages/graduation-awards-sponsorship">Graduation Awards</a></li> <li><a href="/pages/letters-of-support">Letters of Support</a></li> </ul> </div> <div class="col-xs-12 col-sm-3"> <span class="h2">Your Account</span> <ul class="nav-footer"> <li><a href="/account/login">Log In</a></li> <li><a href="/account/register">Create Account</a></li> </ul> <span class="h2">Order Information</span> <ul class="nav-footer"> <li><a href="/pages/how-to-order">How to Order</a></li> <li><a href="/pages/worldwide-shipping">Worldwide Shipping Charges</a></li> <li><a href="/pages/returns-and-cancellations">Returns and Cancellations</a></li> <li><a href="/pages/warranty-information">Warranty Information</a></li> <li><a href="/pages/distributors">Regional Distributors</a></li> <li><a href="/collections/deals-and-discounts">Deals and Discounts</a></li> <li><a href="/pages/price-drop-guarantee">Price Drop Guarantee</a></li> </ul> </div> <div class="col-xs-12 col-sm-3"> <span class="h2">Customer Support</span> <ul class="nav-footer"> <li><svg class="icon icon-envelope"><use xlink:href="#icon-envelope"></use></svg> <a href="/pages/contact-us">Send an Enquiry</a></li> <li><svg class="icon icon-envelope"><use xlink:href="#icon-envelope"></use></svg> <a href="mailto:info@ossila.com">info@ossila.com</a></li> <li class="no-padding-bottom"><a href="https://www.ossila.com/pages/about#our-locations" class="text-bold">Main Office</a></li> <li class="no-padding"><svg class="icon icon-phone"><use xlink:href="#icon-phone"></use></svg> +44 (0)114 2999 180</li> <li class="no-padding-top">Mon-Fri, 8:00-17:00 (GMT/BST)</li> <li class="no-padding-bottom"><a href="https://www.ossila.com/pages/about#our-locations" class="text-bold">EU Office</a></li> <li class="no-padding"><svg class="icon icon-phone"><use xlink:href="#icon-phone"></use></svg> +31 (0)718 081020</li> <li class="no-padding-top">Mon-Fri, 9:00-17:00 (CET/CEST)</li> </ul> </div> </div> <div class="text-center"> <span class="double-right-padding">Ossila Ltd</span> <span class="double-right-padding">Company Number 06920105</span> <span class="double-right-padding">VAT Number GB 978 2092 81</span> <span>EORI Number GB978209281000</span> </div> <div class="text-center"> <span class="double-right-padding">Ossila BV</span> <span class="double-right-padding">CCI number 84102241</span> <span class="double-right-padding">VAT Number NL 863097182B01</span> <span>EORI Number NL863097182</span> </div> </div> </footer> </div> <div id="bottom-banner" class="hidden"> <div id="cookie-banner" class="hidden"> <div class="container"> <div class="row"> <span class="col-xs-12 col-sm-8 no-margin pad-half-height">The Ossila website uses cookies for core functionality. 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