Journal article
Direct determination of amino acids by hydrophilic interaction liquid chromatography with charged aerosol detection
Journal of Chromatography A, v 1446, pp 41-49
13 May 2016
PMID: 27059400
Abstract
•A broad review of amino acid analysis by HPLC.•Novel 18-min separation and quantitation of amino acids without derivatization.•Comparison of quantitative data from UV absorbance and charged aerosol detection.•Quantitation of amino acid content in protein hydrolysates of selected nutritional supplements.
A chromatographic analytical method for the direct determination of amino acids by hydrophilic interaction liquid chromatography (HILIC) was developed. A dual gradient simultaneously varying the pH 3.2 ammonium formate buffer concentration and level of acetonitrile (ACN) in the mobile phase was employed. Using a charged aerosol detector (CAD) and a 2nd order regression analysis, the fit of the calibration curve showed R2 values between 0.9997 and 0.9985 from 1.5mg/mL to 50μg/mL (600ng to 20ng on column). Analyte chromatographic parameters such as the sensitivity of retention to the water fraction in the mobile phase values (mHILIC) were determined as part of method development. A degradation product of glutamine (5-pyrrolidone-2-carboxylic acid; pGlu) was observed and resolved chromatographically with no method modifications. The separation was used to quantitate amino acid content in acid hydrolysates of various protein samples.
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Details
- Title
- Direct determination of amino acids by hydrophilic interaction liquid chromatography with charged aerosol detection
- Creators
- Adam Socia - Merck Research Laboratories, WP78-210, P.O. Box 4, West Point, PA 19486, USAJoe P Foley - Department of Chemistry, Drexel University, 3141 Chestnut Street, Philadelphia, PA 19104, USA
- Publication Details
- Journal of Chromatography A, v 1446, pp 41-49
- Publisher
- Elsevier
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Chemistry
- Web of Science ID
- WOS:000375506700015
- Scopus ID
- 2-s2.0-84979467026
- Other Identifier
- 991014877932104721
InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Collaboration types
- Industry collaboration
- Domestic collaboration
- Web of Science research areas
- Biochemical Research Methods
- Chemistry, Analytical