Spe-uplc Method
Spe-uplc Method - Related Videos
10:24
Non-destructive SPE-UPLC-based Quantification of Aflatoxins and Stilbenoid Phytoalexins in Single Peanut (Arachis spp.) Seeds
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2024
We demonstrate a medium-throughput method for quantification of aflatoxins and stilbenoid phytoalexins in single peanut seeds using ultra performance liquid chromatography. This method was specifically developed for the analyses of wild Arachis species challenged by the aflatoxigenic Aspergillus species.
11:00
Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry (UPLC-HRMS)
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Cited by 33 •
2013
Untargeted metabolomics provides a hypothesis generating snapshot of a metabolic profile. This protocol will demonstrate the extraction and analysis of metabolites from cells, serum, or tissue. A range of metabolites are surveyed using liquid-liquid phase extraction, microflow ultraperformance liquid chromatography/high-resolution mass spectrometry (UPLC-HRMS) coupled to differential analysis software.
07:34
Large Scale Non-targeted Metabolomic Profiling of Serum by Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS)
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Cited by 18 •
2013
Non-targeted metabolite profiling by ultra performance liquid chromatography coupled with mass spectrometry (UPLC-MS) is a powerful technique to investigate metabolism. This article outlines a typical workflow utilized for non-targeted metabolite profiling of serum including sample organization and preparation, data acquisition, data analysis, quality control, and metabolite identification.
10:22
Improved UPLC-UV Method for the Quantification of Vitamin C in Lettuce Varieties (Lactuca sativa L.) and Crop Wild Relatives (Lactuca spp.)
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Cited by 10 •
2020
We present a fast and reliable method to quantify vitamin C in Lactuca spp. using UPLC-UV, potentially transferable to other plants. The key steps are the sample preparation and vitamin C extraction under stable conditions, the reduction of dehydroascorbic acid to ascorbic acid and the optimization of the chromatographic procedure.
09:43
Microfluidic Chip Fabrication and Method to Detect Influenza
0 Views •
Cited by 6 •
2013
An integrated microfluidic thermoplastic chip has been developed for use as a molecular diagnostic. The chip performs nucleic acid extraction, reverse transcriptase, and PCR. Methods for fabricating and running the chip are described.
FAQs
The SPE stage retains target analytes on a sorbent while unwanted matrix components are removed during washing. This cleanup reduces the amount of interfering material that reaches the chromatographic system and concentrates the compounds of interest in a smaller elution volume. As a result, subsequent separation and detection can provide more sensitive and reliable measurements in complex biochemical samples.
UPLC uses high-pressure flow through a column packed with small particles. This configuration separates the enriched sample efficiently, producing rapid resolution of compounds that may be difficult to distinguish in complex mixtures. After SPE has reduced matrix complexity, the UPLC stage can resolve metabolites, peptides, pharmaceuticals, and other biomolecules more effectively for downstream measurement.
SPE primarily prepares the sample by selectively retaining analytes, washing away matrix components, and concentrating the retained material. UPLC then separates the enriched compounds chromatographically. Keeping these roles distinct explains why the combined workflow can outperform either preparation or separation alone: cleanup and enrichment occur before high-resolution separation and instrumental detection.
A typical workflow first exposes the biochemical sample to an SPE sorbent so selected analytes bind. Matrix components are washed away, and an elution solvent transfers the retained analytes into a more concentrated sample. That eluate is then introduced into the UPLC system, where high-pressure column separation prepares the compounds for mass spectrometric or other detection.
The method supports analysis of several important biochemical compound classes, including metabolites, peptides, pharmaceuticals, and other biomolecules. Its value is greatest when these analytes occur within complex samples containing interfering matrix components. SPE provides enrichment and cleanup, while UPLC supplies separation, allowing the resulting workflow to support both compound identification and measurement.
When coupled with mass spectrometric or other detection, the workflow supports qualitative and quantitative measurements. Qualitative analysis can help characterize which compounds are present after cleanup and separation, whereas quantitative analysis determines their measured amounts. Reduced matrix interference and enriched analytes strengthen the reliability of these outcomes in biochemical research.