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Novel approach to determine ghrelin analogs by liquid chromatography with mass spectrometry using a monolithic column
Author(s) -
Zemenova Jana,
Sykora David,
Adamkova Hana,
Maletinska Lenka,
Elbert Tomas,
Marek Ales,
Blechova Miroslava
Publication year - 2017
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201601141
Subject(s) - chromatography , chemistry , mass spectrometry , ghrelin , calibration curve , analytical chemistry (journal) , monolithic hplc column , high performance liquid chromatography , liquid chromatography–mass spectrometry , quantitative analysis (chemistry) , detection limit , biochemistry , hormone
In our project, ghrelin analogs possessing enhanced stability and potential to significantly increase food intake were used. Three newly synthesized ghrelin analogs with fatty acid residues consisting of 8, 10, and 14 carbon atoms were studied. The main goal of this work was to develop a suitable analytical method for the determination of the stability of the novel ghrelin analogs in plasma. An appropriate liquid chromatography‐mass spectrometry method was developed and optimized. The results obtained were compared with the data measured by using a commercial enzyme‐linked immunosorbent assay kit, and a good correlation was found. A preparation strategy for plasma samples was optimized and consisted of simple dilution of the plasma samples followed by direct injection onto a very short monolithic column in combination with mass spectrometric detection. The developed analytical method was utilized for the determination of the stability of the prepared lipopeptides in plasma and for the quantification of the lipopeptides in a preliminary pharmacokinetic study. The feasibility of the developed separation method was clearly demonstrated. Accuracy and precision were within 80–120% and ±20% limits, respectively. Calibration curves were constructed in the range of 1–250 μg/mL.