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Validation of a reverse-phase high-performance liquid chromatography method for the determination of amino acids in gelatins by application of 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate reagent


Citation

Mohd Ismail, Azilawati and Mat Hashim, Dzulkifly and Bakar, Jamilah and Ismail, Amin (2014) Validation of a reverse-phase high-performance liquid chromatography method for the determination of amino acids in gelatins by application of 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate reagent. Journal of Chromatography A, 1353. pp. 49-56. ISSN 0021-9673; ESSN: 1873-3778

Abstract

In-house method validation was conducted to determine amino acid composition in gelatin by a pre-column derivatization procedure with the 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate reagent. The analytical parameters revealed that the validated method was capable of selectively performing a good chromatographic separation for 18 amino acids in less than 40 min; the overall detection and quantitation limit for amino acids fell into ranges of 5.68–12.48 and 36.0–39.0 pmol/μl, respectively; the matrix effect was not observed, and the linearity range was 37.5–1000 pmol/μl. The accuracy (precision and recovery) analyses of the method were conducted under repeatable conditions on different days in random order. Method precision revealed by HorRat values was significantly less than 2, except for histidine with a precision of 2.19, and the method recoveries had a range of 80–115% except for alanine which was recovered at 79.4%. The findings were reproducible and accurately defined, and the method was found to be suited to routine analysis of amino acid composition in gelatin-based ingredients.


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Additional Metadata

Item Type: Article
Divisions: Faculty of Medicine and Health Science
Halal Products Research Institute
DOI Number: https://doi.org/10.1016/j.chroma.2014.04.050
Publisher: Elsevier
Keywords: Gelatin; Amino acid; In-house method validation; 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate reagent
Depositing User: Nurul Ainie Mokhtar
Date Deposited: 12 Feb 2016 02:38
Last Modified: 12 Feb 2016 02:38
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1016/j.chroma.2014.04.050
URI: http://psasir.upm.edu.my/id/eprint/35958
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