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Spectrophotometry: Introduction01:16

Spectrophotometry: Introduction

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Spectrophotometry is the quantitative measurement of the absorption, reflection, diffraction, or transmission of electromagnetic radiation through a material as a function of the intensity and wavelength of the radiation. A spectrophotometer is a device used to measure the change in the radiation intensity caused by its interaction with the material.
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Ultraviolet and Visible (UV–Vis) Spectroscopy: Overview01:02

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Ultraviolet–visible (UV–visible or UV–Vis) spectroscopy is an analytical technique that investigates the interaction between matter and UV–Vis light within the electromagnetic spectrum. This method is widely used for its versatility, simplicity, and relatively quick data acquisition, making it valuable for both qualitative and quantitative analysis. When UV–Vis radiation passes through a material,  molecules absorb light depending on the energy required for...
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UV–Vis Spectroscopy: Beer–Lambert Law01:09

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The Beer-Lambert law describes the relationship between absorbance and concentration, which combines the principles established by scientists Johann Heinrich Lambert and August Beer. Lambert's law states that when light passes through a medium, the loss in intensity is directly proportional to the original intensity and the path length of the light. Beer's law proposed that the transmittance of a solution remains constant if the product of concentration and path length is constant. The modern...
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Organic compounds with conjugated double bonds show strong absorption features in the UV–visible region of the electromagnetic spectrum attributed to π → π* electronic excitations. Generally, a UV–vis absorption spectrum is recorded as a plot of absorbance vs wavelength. The wavelength of maximum absorbance, which manifests as a peak in the absorption spectrum, is denoted as λmax.
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In Ultraviolet–Visible (UV–Vis) spectroscopy, the absorption of electromagnetic radiation is used to probe the electronic structure of molecules. This technique provides insights into molecular electronic transitions, particularly the movement of electrons between different molecular orbitals. Radiation is absorbed if the energy of the electromagnetic radiation passing through the molecule is precisely equal to the energy difference between the excited and ground states. During this...
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UV–Visible absorption spectra of conjugated dienes arise from the lowest energy π → π* transitions. The light-absorbing part of the molecule is called the chromophore, and the substituents directly attached to the chromophore are called auxochromes. A strong correlation exists between the absorption maxima, λmax, and the structure of a conjugated π system. The Woodward–Fieser rules predict the value of λmax for a...
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Related Experiment Video

Updated: May 6, 2026

Preparation of Mica Supported Lipid Bilayers for High Resolution Optical Microscopy Imaging
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Spectral diffusion at the water/lipid interface revealed by two-dimensional fourth-order optical spectroscopy: a

Yuki Nagata1, Shaul Mukamel

  • 1Department of Chemistry, University of California, Irvine, California 92697, United States. nagatay@uci.edu

Journal of the American Chemical Society
|February 19, 2011
PubMed
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We simulated water

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Area of Science:

  • Physical Chemistry
  • Biophysics
  • Spectroscopy

Background:

  • Water's interaction with lipid bilayers is crucial for cell membrane function.
  • Understanding interfacial water dynamics provides insight into biological processes.
  • Phosphatidylcholine bilayers are common models for cell membranes.

Purpose of the Study:

  • To investigate the structure and dynamics of water at the interface of a lipid bilayer.
  • To differentiate between bulk-like and interfacial water environments using nonlinear spectroscopy.
  • To explore coherence transfer and spectral diffusion in interfacial water.

Main Methods:

  • Classical simulation protocol for nonlinear optical signals.
  • Three infrared (IR) probe pulses followed by one visible probe pulse.
  • Sum-frequency-generation (SFG) 1D and 2D spectroscopy.

Main Results:

  • Two distinct OH stretch environments identified: near-bulk and top-layer water adjacent to DMPC.
  • Asymmetric cross-peak pattern in 2D spectra indicates coherence transfer between water modes.
  • Near-bulk water exhibits fast spectral diffusion, similar to bulk water.
  • Top-layer water shows slower spectral diffusion due to DMPC interaction.

Conclusions:

  • The study reveals distinct dynamical behaviors of water layers near a DMPC bilayer.
  • Nonlinear optical spectroscopy effectively distinguishes interfacial water from bulk water.
  • Coherent energy transfer occurs between different water vibrational modes at the interface.