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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Chuanxiang, Wu Lian, Xia Lijie, Liu Fengli, Qu Zhiwei, Sun Xianen, Zhao Jinmao, You |
| Description | Author Affiliation: Chuanxiang W ( Key Laboratory of Life-Organic Analysis of Shandong Province, Qufu Normal University, Qufu, PR China.); Lian X ( Key Laboratory of Life-Organic Analysis of Shandong Province, Qufu Normal University, Qufu, PR China. Electronic address: xialian01@163.com.); Lijie L ( Key Laboratory of Life-Organic Analysis of Shandong Province, Qufu Normal University, Qufu, PR China.); Fengli Q ( Key Laboratory of Life-Organic Analysis of Shandong Province, Qufu Normal University, Qufu, PR China.); Zhiwei S ( Key Laboratory of Life-Organic Analysis of Shandong Province, Qufu Normal University, Qufu, PR China.); Xianen Z ( Key Laboratory of Life-Organic Analysis of Shandong Province, Qufu Normal University, Qufu, PR China.); Jinmao Y ( Key Laboratory of Life-Organic Analysis of Shandong Province, Qufu Normal University, Qufu, PR China) |
| Abstract | A sensitive and efficient method of high performance liquid chromatography using 1-(2-naphthyl)-3-methyl-5-pyrazolone (NMP) as pre-column derivatization reagent coupled with UV detection (HPLC-UV) and online mass spectrometry identification was established for determination of the most common N-Acetylhexosamines (N-acetyl-d-glucosamine (GlcNAc) and N-acetyl-d-galactosamine (GalNAc)) and N-acetylneuraminic acid (Neu5Ac). In order to obtain the highest liberation level of the three monosaccharides without destruction of Neu5Ac or conversion of GlcNAc/GalNAc to GlcN/GalN in the hydrolysis procedure, the pivotal parameters affecting the liberation of N-acetylhexosamines/Neu5Ac from sample were investigated with response surface methodology (RSM). Under the optimized condition, maximum yield was obtained. The effects of key parameters on derivatization, separation and detection were also investigated. At optimized conditions, three monosaccharides were labeled fast and entirely, and all derivatives exhibited a good baseline resolution and high detection sensitivity. The developed method was linear over the calibration range 0.25-12µM, with R(2)>0.9991. The detection limits of the method were between 0.48 and 2.01pmol. Intra- and inter-day precisions for the three monosaccharides (GlcNAc, GalNAc and Neu5Ac) were found to be in the range of 3.07-4.02% and 3.69-4.67%, respectively. Individual monosaccharide recovery from spiked milk was in the range of 81%-97%. The sensitivity of the method, the facility of the derivatization procedure and the reliability of the hydrolysis conditions suggest the proposed method has a high potential for utilization in routine trace N-acetylhexosamines and Neu5Ac analysis in biological samples. |
| File Format | HTM / HTML |
| ISSN | 15700232 |
| Volume Number | 1011 |
| e-ISSN | 1873376X |
| Journal | Journal of Chromatography B |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2016-02-01 |
| Publisher Place | Netherlands |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Analytical Chemistry Chromatography, High Pressure Liquid Methods Hexosamines Analysis Milk, Human Chemistry N-acetylneuraminic Acid Tandem Mass Spectrometry Isolation & Purification Humans Limit Of Detection Linear Models Reproducibility Of Results Spectrophotometry, Ultraviolet Journal Article Research Support, Non-u.s. Gov't |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Medicine Analytical Chemistry Clinical Biochemistry Biochemistry |
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