title: Development and validation of a HPLC method for determination of degree of polymerization of xylo-oligosaccharides.
abstract: A reliable reversed-phase HPLC method was developed for high resolution separation and high sensitivity determination of xylo-oligosaccharides (XOS) with degree of polymerization from 2 to 8. The method was carried out on a Kromasil C18 column using pre-column derivatization with 1-phenyl-3-methyl-5-pyrazolone (PMP) and UV detection at 245nm. The effects of pH value of mobile phase, volume proportion of acetonitrile, concentration of ammonium acetate buffer and flow rate on the retention time and degree of separation of XOS derivatives were investigated. A satisfactory result was achieved in 25min with a mobile phase of 10mmol/L ammonium acetate buffer (pH5.5)-acetonitrile by a gradient elution at 0.8mL/min. In addition, this method was validated by liquid chromatography-tandem mass spectroscopy (LC-MS) analysis and several uncertain compounds were identified. The proposed HPLC method is suitable for the compositional analysis and quality control of XOS.
authors.affiliation: Key Laboratory of Marine Drugs, Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, China; Shandong Provincial Key laboratory of Glycoscience and Glycoengineering, Ocean University of China, Qingdao 266003, China.
authors.forename: Jianghua
authors.initials: J
authors.lastname: Pu
authors.affiliation: Key Laboratory of Marine Drugs, Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, China; Shandong Provincial Key laboratory of Glycoscience and Glycoengineering, Ocean University of China, Qingdao 266003, China. Electronic address: zhaoxia@ouc.edu.cn.
authors.forename: Xia
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Key Laboratory of Marine Drugs, Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, China; Shandong Provincial Key laboratory of Glycoscience and Glycoengineering, Ocean University of China, Qingdao 266003, China.
authors.forename: Qingchi
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Key Laboratory of Marine Drugs, Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, China.
authors.forename: Yingdi
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Key Laboratory of Marine Drugs, Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhou
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000592:standards / Q000639:trends; D005965:Glucuronates / Q000032:analysis*; D009844:Oligosaccharides / Q000032:analysis*; D058105:Polymerization; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000592:standards* / Q000639:trends
chemical_list: D005965:Glucuronates; D009844:Oligosaccharides; C570991:xylooligosaccharide
keywords: 1-Phenyl-3-methyl-5-pyrazolone; Degree of polymerization; HPLC; LC–MS; Pre-column derivatization; Xylo-oligosaccharide