title: Validated HPAEC-PAD Method for the Determination of Fully Deacetylated Chitooligosaccharides.
abstract: An efficient and sensitive analytical method based on high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) was established for the simultaneous separation and determination of glucosamine (GlcN)₁ and chitooligosaccharides (COS) ranging from (GlcN)₂ to (GlcN)₆ without prior derivatization. Detection limits were 0.003 to 0.016 mg/L (corresponding to 0.4-0.6 pmol), and the linear range was 0.2 to 10 mg/L. The optimized analysis was carried out on a CarboPac-PA100 analytical column (4 × 250 mm) using isocratic elution with 0.2 M aqueous sodium hydroxide-water mixture (10:90,v/v) as the mobile phase at a 0.4 mL/min flow rate. Regression equations revealed a good linear relationship (R² = 0.9979-0.9995,n= 7) within the test ranges. Quality parameters, including precision and accuracy, were fully validated and found to be satisfactory. The fully validated HPAEC-PAD method was readily applied for the quantification of (GlcN)1-6in a commercial COS technical concentrate. The established method was also used to monitor the acid hydrolysis of a COS technical concentrate to ensure optimization of reaction conditions and minimization of (GlcN)₁ degradation.
authors.affiliation: Key Laboratory of Pesticide Chemistry and Application, Ministry of Agriculture, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, No. 2 Yuanmingyuan West Road, Beijing 100193, China. caolidong@caas.cn.
authors.forename: Lidong
authors.initials: L
authors.lastname: Cao
authors.affiliation: Institute for the Control of Agrochemicals, Ministry of Agriculture, No. 22 Maizidian Street, Beijing 110000, China. wjl_icama@126.com.
authors.forename: Jinlong
authors.initials: J
authors.lastname: Wu
authors.affiliation: Key Laboratory of Pesticide Chemistry and Application, Ministry of Agriculture, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, No. 2 Yuanmingyuan West Road, Beijing 100193, China. lixiuhuan0822@163.com.
authors.forename: Xiuhuan
authors.initials: X
authors.lastname: Li
authors.affiliation: Key Laboratory of Pesticide Chemistry and Application, Ministry of Agriculture, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, No. 2 Yuanmingyuan West Road, Beijing 100193, China. zhengli7seven@163.com.
authors.forename: Li
authors.initials: L
authors.lastname: Zheng
authors.affiliation: Key Laboratory of Pesticide Chemistry and Application, Ministry of Agriculture, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, No. 2 Yuanmingyuan West Road, Beijing 100193, China. wumiaomiao2016@163.com.
authors.forename: Miaomiao
authors.initials: M
authors.lastname: Wu
authors.affiliation: Institute for the Control of Agrochemicals, Ministry of Agriculture, No. 22 Maizidian Street, Beijing 110000, China. liupingping@agri.gov.cn.
authors.forename: Pingping
authors.initials: P
authors.lastname: Liu
authors.affiliation: Key Laboratory of Pesticide Chemistry and Application, Ministry of Agriculture, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, No. 2 Yuanmingyuan West Road, Beijing 100193, China. qlhuang@ippcaas.cn.
authors.forename: Qiliang
authors.initials: Q
authors.lastname: Huang
mesh_terms: D002138:Calibration; D002623:Chemistry Techniques, Analytical / Q000379:methods* / Q000592:standards; D002686:Chitin / Q000031:analogs & derivatives* / Q000032:analysis / Q000592:standards; D048271:Chitosan; D002851:Chromatography, High Pressure Liquid / Q000592:standards; D055664:Electrochemical Techniques / Q000592:standards; D005944:Glucosamine / Q000032:analysis; D057230:Limit of Detection; D009844:Oligosaccharides; D012972:Sodium Hydroxide / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014867:Water / Q000737:chemistry
chemical_list: D009844:Oligosaccharides; C493484:oligochitosan; D014867:Water; D002686:Chitin; D012972:Sodium Hydroxide; D048271:Chitosan; D005944:Glucosamine
keywords: HPAEC-PAD; chitooligosaccharides; hydrolysis; method validation; quantification