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Evaluation of amino acids in Aminoven 10 % and Vamin 18EF with UPLC using Waters AccQTag method
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
2013 (English)Independent thesis Advanced level (professional degree), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

The aim with this project was to evaluate Waters AccQTag method which was ananalytical method that Waters Corporation provides for determination of aminoacids. The amino acid analysis was carried out by reversed-phase AQUITY UPLCH-Class, using a BEH C18-column and a PDA detector.Both primary and secondary amino acids were analyzed after precolumnderivatization with the AccQTag reagent kit by a manual procedure and, detected at260 nm. Fresenius Kabis existing products, Aminoven 10 % and Vamin 18EF wereused as samples. The evaluation was based on external calibration curves withprepared standard solutions from Fresenius Kabi, with the addition of the internalstandard L-Norvaline. Waters amino acid standard was used for the system suitabilitytest. A validation according to the ICH guidelines were performed with the followingparameters: specificity, linearity, precision, LOQ, LOD and robustness.Complementary parameters for system suitability test (SST) and stability of standardand reagent were also evaluated. The validation showed that the % RSD values forspecificity, system suitability test and robustness were within the acceptance criteriaof equal or less than 3 % for amino acids. Although the precision were between1.3-7.5 % which was higher than the acceptance criteria of equal or less than 3 %, thiswas likely due to sensitive sample preparation. The linearity was bad which made itimpossible to quantify the method since the calibration curves were below 0.999. Itwas concluded that the system was robust and can be used to analyze amino acids inAminoven 10 % and Vamin 18EF but the preparation step should if possible beautomatized to achieve better precision in the analytical work. Two amino acids,cysteine and histidine, needs to be studied further; cysteine gave three peaks insteadof one and histidine gave a peak split at higher injection volume than 0.5 uL.

Place, publisher, year, edition, pages
2013. , 25 p.
UPTEC K, ISSN 1650-8297 ; 12023
Keyword [en]
Analytical chemistry, UPLC, amino acids
National Category
Pharmaceutical Chemistry
URN: urn:nbn:se:uu:diva-194120OAI: diva2:604347
External cooperation
Fresenius Kabi
Educational program
Master Programme in Chemical Engineering
Available from: 2013-03-11 Created: 2013-02-09 Last updated: 2013-03-11Bibliographically approved

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