Related Experiment Video
Updated: Jan 30, 2026
14:12
HPLC Measurement of the DNA Oxidation Biomarker, 8-oxo-7,8-dihydro-2’-deoxyguanosine, in Cultured Cells and Animal Tissues
Published on: August 1, 2015
27.6K
HPLC-UV-MS Analysis: A Source for Severe Oxidation Artifacts
Fritz Schweikart1, Gustaf Hulthe2
1Advanced Drug Delivery, Pharmaceutical Sciences, IMED Biotech Unit , AstraZeneca , Gothenburg 43183 , Sweden.
Analytical Chemistry
|January 24, 2019
Summary
UV detectors in HPLC systems can cause unwanted artifacts in mass spectrometry (MS) analysis. Radicals formed by UV light interfere with low-concentration samples, leading to misleading results in pharmaceutical development.
Area of Science:
- Analytical Chemistry
- Pharmaceutical Analysis
- Chromatography
Background:
- High-Performance Liquid Chromatography (HPLC) coupled with UV and Mass Spectrometry (MS) is standard for purity assessments.
- Advancements in instrument sensitivity necessitate lower sample concentrations.
- Modern UV detectors exhibit higher light flux compared to previous generations.
Purpose of the Study:
- To investigate the impact of UV detector-induced degradation on MS signal integrity.
- To identify and illustrate artifact generation in typical pharmaceutical development samples.
- To highlight the potential for misleading mass spectra due to UV photodegradation.
Main Methods:
- Utilized HPLC coupled to UV and MS detection.
- Analyzed pharmaceutical development samples with varying analyte concentrations.
- Observed and characterized artifact signals in MS spectra resulting from UV exposure within the detector.
Main Results:
- UV light in the detector generates sufficient radicals to interfere with low analyte levels.
- Artifact signals in MS spectra were observed and directly linked to UV degradation.
- Examples from pharmaceutical development demonstrate severely misleading mass spectra.
Conclusions:
- Photodegradation within UV detectors poses a significant challenge for accurate MS analysis.
- The high light flux in modern UV detectors exacerbates this issue, especially with sensitive MS instruments.
- Careful consideration of UV detector use is crucial to avoid artifactual data in purity assessments.
Related Concept Videos
01:11Nodal Analysis with Voltage Sources
2.0K
Nodal analysis is a remarkably effective method used in electrical engineering to simplify the analysis of complex circuits, including those with dependent or independent voltage sources. Its strength lies in its systematic approach to breaking down circuits into manageable components, making it easier for engineers to understand and solve.
Consider a circuit that contains four resistors and two voltage sources, as shown in Figure 1. One of these voltage sources is connected between a...
Consider a circuit that contains four resistors and two voltage sources, as shown in Figure 1. One of these voltage sources is connected between a...
2.0K
01:10Mesh Analysis with Current Sources
2.0K
Mesh analysis becomes simpler when analyzing circuits with current sources, whether independent or dependent. The presence of current sources reduces the number of equations required for analysis. Two cases illustrate this:
Current Source in One Mesh: The analysis process is straightforward when a current source is found in only one mesh within the circuit. Mesh currents are assigned as usual, with the mesh containing the current source excluded from the analysis. Kirchhoff's voltage law...
Current Source in One Mesh: The analysis process is straightforward when a current source is found in only one mesh within the circuit. Mesh currents are assigned as usual, with the mesh containing the current source excluded from the analysis. Kirchhoff's voltage law...
2.0K
03:14Oxidation Numbers
42.6K
In redox reactions, the transfer of electrons occurs between reacting species. Electron transfer is described by a hypothetical number called the oxidation number (or oxidation state). It represents the effective charge of an atom or element, which is assigned using a set of rules.
42.6K
01:15Pyruvate Oxidation
168.8K
After glycolysis, the charged pyruvate molecules enter the mitochondria via active transport and undergo three enzymatic reactions. These reactions ensure that pyruvate can enter the next metabolic pathway so that energy stored in the pyruvate molecules can be harnessed by the cells.
First, the enzyme pyruvate dehydrogenase removes the carboxyl group from pyruvate and releases it as carbon dioxide. The stripped molecule is then oxidized and releases electrons, which are then picked up by NAD+...
First, the enzyme pyruvate dehydrogenase removes the carboxyl group from pyruvate and releases it as carbon dioxide. The stripped molecule is then oxidized and releases electrons, which are then picked up by NAD+...
168.8K
03:11Oxidation-Reduction Reactions
75.6K
Oxidation–Reduction Reactions
75.6K
01:30UV–Vis Spectrum
2.1K
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...
The UV–Vis spectrum of a molecule is the plot of its absorbance versus wavelength. The plot is drawn by taking molar...
2.1K