Topic
Elution
About: Elution is a research topic. Over the lifetime, 18974 publications have been published within this topic receiving 386005 citations.
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TL;DR: An improved method for extracting the lipids from tissues consists of the use of hexane:isopropanol, followed by a wash of the extract with aqueous sodium sulfate to remove nonlipid contaminants.
2,400 citations
TL;DR: The combination of strong cation exchange (SCX) and reversed-phase (RP) chromatography to achieve two-dimensional separation prior to MS/MS and 1,504 yeast proteins were unambiguously identified in this single analysis.
Abstract: Highly complex protein mixtures can be directly analyzed after proteolysis by liquid chromatography coupled with tandem mass spectrometry (LC−MS/MS). In this paper, we have utilized the combination of strong cation exchange (SCX) and reversed-phase (RP) chromatography to achieve two-dimensional separation prior to MS/MS. One milligram of whole yeast protein was proteolyzed and separated by SCX chromatography (2.1 mm i.d.) with fraction collection every minute during an 80-min elution. Eighty fractions were reduced in volume and then re-injected via an autosampler in an automated fashion using a vented-column (100 μm i.d.) approach for RP-LC−MS/MS analysis. More than 162 000 MS/MS spectra were collected with 26 815 matched to yeast peptides (7537 unique peptides). A total of 1504 yeast proteins were unambiguously identified in this single analysis. We present a comparison of this experiment with a previously published yeast proteome analysis by Yates and colleagues (Washburn, M. P.; Wolters, D.; Yates, J. ...
1,654 citations
TL;DR: All amino acids, including proline, are converted quantitatively to phenylthiocarbamyl compounds and these are stable enough to eliminate any need for in-line derivatization, providing results comparable in sensitivity and precision to those obtained by state-of-the-art ion-exchange analyzers.
1,516 citations
TL;DR: Methods for preparation of stable columns of calcium phosphate (hydroxylapatite) suitable for protein chromatography and conditions for specific elution of proteins on such columns have been worked out, particularly the narrow elution ranges and the marked mutual displacement effects shown by many proteins.
1,341 citations
TL;DR: In this article, the application of multivariate curve resolution to second order data from hyphenated liquid chromatography with spectrometric diode array detection is shown, where the authors apply it to second-order data with different data structures depending on the reproducibility of the elution process.
1,208 citations