Home|Journals|Articles by Year|Audio Abstracts
 

Original Article



Use of HPLC-UV method for the analysis of maple syrup urine disease in plasma sample first time in Saudi Arabia

Abdul Rafiq Khan, Ali Al-Othaim, Ahmed Al-Fares, Najla Al Hussain.




Abstract

Background: Measurement of branched-chain amino acids (BCAAs) [valine (Val), alloisoleucine (allo-Ile), isoleucine (Ile), and leucine (Leu)] in plasma and dry blood spot samples is important for the diagnosis and monitoring of maple syrup urine disease (MSUD), which is a metabolic disorder that affects metabolism of BCAAs. We used a temporally short program on an High performance liquid chromatography (HPLC) (Biochrom 30) for the first time in Saudi Arabia to analyze plasma samples for the diagnosis of MSUD.
Methodology: 500 μl of plasma were mixed with 500 μl of 10% sulphosalicylic acid, followed by shaking for 5-10 seconds on a vortexer. The mixture was incubated at 4 C for 10 minutes followed by centrifugation at 10,000 rpm for 5 minutes. Supernatants were transferred into HPLC vials for injection.
Results: For all amino acids, the limit of detection and limit of quantification were 5.0 and 10.0 μmol/l, respectively. The method was linear in the range of 10-2,500 μmol/l. The method was specific and selective for the detection of BCAAs, including alloisoleucine. The correlation coefficient (CV) of method comparison was greater than 0.980 and percent recovery of all amino acids in plasma was 80%-100%, which was evaluated at two amino acid concentrations. Precision was assessed by repeat analysis of spiked plasma samples and percent CV was found to be

Key words: MSUD HPLC analysis






Full-text options


Share this Article


Online Article Submission
• ejmanager.com




ejPort - eJManager.com
Refer & Earn
JournalList
About BiblioMed
License Information
Terms & Conditions
Privacy Policy
Contact Us

The articles in Bibliomed are open access articles licensed under Creative Commons Attribution 4.0 International License (CC BY), which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.