Separation and determination of sugars by reversed-phase high-performance liquid chromatography after pre-column microwave-assisted derivatization

• Published in: Analytical and bioanalytical chemistry Vol. 384; no. 3; pp. 844 - 850
• Main Authors: Momenbeik, Fariborz, Khorasani, Jafar H
• Format: Journal Article
• Language: English
• Published: Germany Berlin/Heidelberg : Springer-Verlag 01.02.2006
• Subjects: Doehlert design; high performance liquid chromatography; Microwave-assisted reaction; p-Nitroaniline; sugars
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-005-0207-2

In this study the possibility of derivatizing sugars using microwave irradiation was investigated. The amount of reagent, irradiation intensity, and derivatization time were optimized. In the derivatization of sugars with p-nitroaniline the reaction is complete within 5 min at 600 W when the p-nitroaniline-to-sugar and NaBH₃CN-to-sugar mole ratios were above 1.4 and 3.1, respectively. A Doehlert design was used to optimize the mobile phase for separation of p-nitroaniline-labeled sugars; and the best separation was obtained by use of 0.01 mol L-¹ acetate buffer at pH 4.40 containing 11.0% acetonitrile. Analysis using this method was highly sensitive and analysis time was short. Finally, a food sample was analyzed using the proposed method.