| Title |
Improved LC-MS/MS method for the quantification of hepcidin-25 in clinical samples
|
|---|---|
| Published in |
Analytical & Bioanalytical Chemistry, April 2018
|
| DOI | 10.1007/s00216-018-1056-0 |
| Pubmed ID | |
| Authors |
Ioana M. Abbas, Holger Hoffmann, María Montes-Bayón, Michael G. Weller |
| Abstract |
Mass spectrometry-based methods play a crucial role in the quantification of the main iron metabolism regulator hepcidin by singling out the bioactive 25-residue peptide from the other naturally occurring N-truncated isoforms (hepcidin-20, -22, -24), which seem to be inactive in iron homeostasis. However, several difficulties arise in the MS analysis of hepcidin due to the "sticky" character of the peptide and the lack of suitable standards. Here, we propose the use of amino- and fluoro-silanized autosampler vials to reduce hepcidin interaction to laboratory glassware surfaces after testing several types of vials for the preparation of stock solutions and serum samples for isotope dilution liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS). Furthermore, we have investigated two sample preparation strategies and two chromatographic separation conditions with the aim of developing a LC-MS/MS method for the sensitive and reliable quantification of hepcidin-25 in serum samples. A chromatographic separation based on usual acidic mobile phases was compared with a novel approach involving the separation of hepcidin-25 with solvents at high pH containing 0.1% of ammonia. Both methods were applied to clinical samples in an intra-laboratory comparison of two LC-MS/MS methods using the same hepcidin-25 calibrators with good correlation of the results. Finally, we recommend a LC-MS/MS-based quantification method with a dynamic range of 0.5-40 μg/L for the assessment of hepcidin-25 in human serum that uses TFA-based mobile phases and silanized glass vials. Graphical abstract Structure of hepcidin-25 (Protein Data Bank, PDB ID 2KEF). |
X Demographics
The data shown below were collected from the profiles of 6 X users who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| France | 1 | 17% |
| Germany | 1 | 17% |
| Mexico | 1 | 17% |
| United States | 1 | 17% |
| Unknown | 2 | 33% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Scientists | 3 | 50% |
| Members of the public | 2 | 33% |
| Science communicators (journalists, bloggers, editors) | 1 | 17% |
Mendeley readers
The data shown below were compiled from readership statistics for 24 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 24 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 5 | 21% |
| Student > Ph. D. Student | 4 | 17% |
| Other | 3 | 13% |
| Lecturer | 1 | 4% |
| Student > Doctoral Student | 1 | 4% |
| Other | 3 | 13% |
| Unknown | 7 | 29% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 4 | 17% |
| Medicine and Dentistry | 4 | 17% |
| Biochemistry, Genetics and Molecular Biology | 3 | 13% |
| Pharmacology, Toxicology and Pharmaceutical Science | 2 | 8% |
| Chemistry | 2 | 8% |
| Other | 2 | 8% |
| Unknown | 7 | 29% |
Attention Score in Context
This research output has an Altmetric Attention Score of 2. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 25 May 2018.
All research outputs
#15,745,807
of 25,382,440 outputs
Outputs from Analytical & Bioanalytical Chemistry
#4,752
of 9,619 outputs
Outputs of similar age
#190,665
of 340,784 outputs
Outputs of similar age from Analytical & Bioanalytical Chemistry
#65
of 173 outputs
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So far Altmetric has tracked 9,619 research outputs from this source. They receive a mean Attention Score of 3.1. This one is in the 48th percentile – i.e., 48% of its peers scored the same or lower than it.
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