Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Spectrophotometry: Introduction01:16

Spectrophotometry: Introduction

3.0K
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.
The essential components of a spectrophotometer include a source of electromagnetic radiation, a slot for placing a material to be analyzed, and a...
3.0K
UV–Vis Spectrometers01:14

UV–Vis Spectrometers

1.3K
The absorbance of UV and visible (UV–visible) radiations is measured using a UV–visible spectrophotometer. Deuterium lamps, which emit UV radiation, and tungsten lamps, which produce radiation in the visible region, are used as light sources in UV–visible spectrophotometers. A monochromator or prism is used for diffraction grating, i.e., to split the incoming radiation into different wavelengths. A system of slits is used to focus the desired wavelength on the sample cell.
1.3K
Ultraviolet and Visible (UV–Vis) Spectroscopy: Overview01:02

Ultraviolet and Visible (UV–Vis) Spectroscopy: Overview

2.5K
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...
2.5K
Atomic Absorption Spectroscopy: Interference01:25

Atomic Absorption Spectroscopy: Interference

669
Interference leads to systematic error in atomic absorption (AA) measurements by enhancing or diminishing the analytical signal or the background. These interferences can be grouped into three main categories: spectral interference, chemical interference, and physical interference.
Spectral interference occurs when signals from other elements or molecules overlap with the analyte signal, falsely elevating or masking the analyte's absorbance. This interference can be corrected using Zeeman,...
669
UV–Vis Spectroscopy: Beer–Lambert Law01:09

UV–Vis Spectroscopy: Beer–Lambert Law

2.2K
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...
2.2K
UV–Vis Spectrum01:30

UV–Vis Spectrum

1.1K
When light passes through a substance, a portion of the light is absorbed while the remaining light is reflected or transmitted. If the molecule absorbs light between the wavelengths of 180–400 nm range, the UV spectrum is obtained, and if it absorbs light in the 400–780 nm wavelength range, the visible spectrum is obtained.     
The UV–Vis spectrum of a molecule is the plot of its absorbance versus wavelength. The plot is drawn by taking molar...
1.1K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

At the Exposome's Frontier: Mass Spectrometry to Bridge the Gap between Exposure and Biological Mechanisms.

Environmental science & technology·2026
Same author

Gender shapes the relationship between productivity and journal prestige in science.

Scientific reports·2026
Same author

Comprehensive indicators and fine granularity refine density scaling laws in rural-urban systems.

Scientific reports·2026
Same author

Ground-Based Remote Sensing and Machine Learning for in Situ and Noninvasive Monitoring and Identification of Salts and Moisture in Historic Buildings.

Analytical chemistry·2025
Same author

Prediction of Retention Indices in LC-HRMS for Enhanced Structural Identification of Organic Micropollutants in Water: Selectivity-Based Filtration.

Analytical chemistry·2025
Same author

Physicochemical modelling of the retention mechanism of temperature-responsive polymeric columns for HPLC through machine learning algorithms.

Journal of cheminformatics·2024

Related Experiment Video

Updated: Jun 6, 2025

Characterization of Biological Absorption Spectra Spanning the Visible to the Short-Wave Infrared
07:38

Characterization of Biological Absorption Spectra Spanning the Visible to the Short-Wave Infrared

Published on: January 10, 2025

1.1K

Raw data and noise in spectrophotometry.

Bruna Falgueras Vallbona1, Ardiana Kajtazi2, Golnaz Shahtahmassebi3

  • 1Department of Chemistry and Forensics, School of Science and Technology, Nottingham Trent University, Nottingham, NG11 8NS, United Kingdom.

Analytica Chimica Acta
|November 30, 2024
PubMed
Summary
This summary is machine-generated.

Spectrophotometer precision limits are often misunderstood. This study reveals current guidance is outdated, showing optimal performance can exceed traditional absorbance ranges and urging instrument makers for better data transparency.

Keywords:
Dispersion modelFluctuation scalingSpectrophotometer

More Related Videos

Author Spotlight: Exploring Light-Driven Chemical Reactions and Energy-Harnessing Devices in Photochemical Research
08:12

Author Spotlight: Exploring Light-Driven Chemical Reactions and Energy-Harnessing Devices in Photochemical Research

Published on: February 16, 2024

8.5K
Proton Transfer and Protein Conformation Dynamics in Photosensitive Proteins by Time-resolved Step-scan Fourier-transform Infrared Spectroscopy
10:03

Proton Transfer and Protein Conformation Dynamics in Photosensitive Proteins by Time-resolved Step-scan Fourier-transform Infrared Spectroscopy

Published on: June 27, 2014

17.9K

Related Experiment Videos

Last Updated: Jun 6, 2025

Characterization of Biological Absorption Spectra Spanning the Visible to the Short-Wave Infrared
07:38

Characterization of Biological Absorption Spectra Spanning the Visible to the Short-Wave Infrared

Published on: January 10, 2025

1.1K
Author Spotlight: Exploring Light-Driven Chemical Reactions and Energy-Harnessing Devices in Photochemical Research
08:12

Author Spotlight: Exploring Light-Driven Chemical Reactions and Energy-Harnessing Devices in Photochemical Research

Published on: February 16, 2024

8.5K
Proton Transfer and Protein Conformation Dynamics in Photosensitive Proteins by Time-resolved Step-scan Fourier-transform Infrared Spectroscopy
10:03

Proton Transfer and Protein Conformation Dynamics in Photosensitive Proteins by Time-resolved Step-scan Fourier-transform Infrared Spectroscopy

Published on: June 27, 2014

17.9K

Area of Science:

  • Analytical Chemistry
  • Spectroscopy
  • Instrument Science

Background:

  • Spectrophotometer precision limits are under-appreciated, with current guidance based on outdated instruments.
  • Modern spectrophotometers use digitized intensities, differing from historical analog outputs.
  • Assessing modern spectrophotometer limitations is challenging without direct access to raw digitized intensities.

Purpose of the Study:

  • To analyze spectrophotometer performance using derived data when raw intensities (I and I0) are unavailable.
  • To evaluate the validity of current International Union of Pure and Applied Chemistry (IUPAC) guidance on absorbance ranges.
  • To characterize noise and determine optimal performance spectra for modern UV-Vis spectrophotometers.

Main Methods:

  • Analysis of spectrophotometer signals using absorbance and transmittance data.
  • Characterization of noise in UV-Vis spectrophotometers through three distinct methods.
  • Evaluation of relative standard deviation (RSD) across various absorbance values.

Main Results:

  • Current IUPAC guidance restricting absorbance to 0.1-1.0 a.u. lacks empirical justification.
  • Optimal performance (minimum RSD) was not consistently found within the 0.1-1.0 a.u. range.
  • UV-Vis spectrophotometers tested were not Poisson optimal, with best RSDs sometimes exceeding 1.0 a.u.

Conclusions:

  • Classical theories are insufficient for accurately describing modern spectrophotometers.
  • There is a need for IUPAC to update its guidance with current instrument data.
  • Improved data transparency from instrument manufacturers is crucial for optimal spectrophotometer use.