title: Novel reagents for quantitative analysis of valiolamine in biological samples by high-performance liquid chromatography with pre-column UV derivatization.
abstract: A rapid, low-cost, high sensitive and quantitative method to detect valiolamine in a medium for microbial culture, involving derivatization with a new labeling reagent, 4-methoxybenzenesulfonyl fluoride (MOBS-F), followed by reverse-phase high-performance liquid chromatography with ultraviolet (UV) detection with simple operation procedure. 4-Methoxybenzenesulfonyl chloride (MOBS-Cl) and 2-nitrobenzenesulfonyl chloride (NBS-Cl) were compared with MOBS-F as novel reagents in this paper, and the MOBS-F was chosen as the most suitable derivatization reagent. The column was thermostatic at 35 degrees C, the mobile phase flow-rate was 1.0mL/min and the detection wavelength was 240nm. For a biological sample, the separation of the derivatives was achieved using a gradient mobile system. The elution program is 88% phosphate buffer (50mM; pH=3.0) and 12% methanol for 23min, then 70% of phosphate buffer and 30% methanol for another 15min and finally 88% of phosphate buffer and 12% of methanol for 5min to re-equilibrate the column. The optimized conditions of the derivatization were as follows: derivatization reaction temperature 30 degrees C; derivatization reaction pH value 11.0, reaction time 10min and MOBS-F concentration higher than 1.5mg/mL for standard solutions and higher than 5.0mg/mL for the biological sample. Calibration curves were linear in the range of 0.050-25microg/mL for the standard solutions and 1.0-75microg/mL for the biological sample. The sensitive analytical method is helpful to control the biotechnological process of voglibose production and product quality control.
authors.affiliation: Department of Chemistry and Chemical Engineering, Yangzhou University, Yangzhou, China. wangcy@yzu.edu.cn
authors.forename: Chengyin
authors.initials: C
authors.lastname: Wang
authors.forename: Yujie
authors.initials: Y
authors.lastname: Sun
authors.forename: Qunyin
authors.initials: Q
authors.lastname: Wen
authors.forename: Guoxiu
authors.initials: G
authors.lastname: Wang
authors.forename: Yang
authors.initials: Y
authors.lastname: Wang
authors.forename: Qishu
authors.initials: Q
authors.lastname: Qu
authors.forename: Gongjun
authors.initials: G
authors.lastname: Yang
authors.forename: Xiaoya
authors.initials: X
authors.lastname: Hu
mesh_terms: D001709:Biotechnology / Q000379:methods; D002138:Calibration; D002623:Chemistry Techniques, Analytical; D002845:Chromatography / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007294:Inositol / Q000031:analogs & derivatives / Q000032:analysis / Q000737:chemistry; D008956:Models, Chemical; D009578:Nitrobenzenes / Q000032:analysis; D004364:Pharmaceutical Preparations / Q000737:chemistry; D011786:Quality Control; D013696:Temperature; D013997:Time Factors; D014466:Ultraviolet Rays; D050111:Imino Sugars
chemical_list: D007294:Inositol; D009578:Nitrobenzenes; D004364:Pharmaceutical Preparations; C002732:2-nitrobenzenesulfenyl chloride; C043270:valiolamine; D050111:Imino Sugars; C102817:voglibose