ABSTRACT
Water-soluble polysaccharides from traditional Chinese medicine have properties of complex structure and high molecular, resulting in hardly complete their structural characterization.However, a "bottom-up" approach could solve this problem.Glehniae Radix extract was extracted with hot water and then precipitated by 40% ethanol to obtain Glehniae Radix polysaccharides (RGP). Subsequently, a partial acid hydrolysis method was carried out and the effects of acid concentration, time and temperature on hydrolysis were investigated. Under the optimum hydrolysis condition (1.5 mol•L⁻¹ trifluoroacetic acid, 4 h, and 80 ℃), RGP were hydrolyzed to characteristic oligosaccharide fragments. Futher, a hydrophilic liquid chromatography- mass spectrometry method was used for the separation and structural characterization of the polysaccharide hydrolysates. According to MS and MS/MS analysis of several standard disaccharides, a method for determining the type of polysaccharide glycosidic linkage by mass spectrometry was established. The results showed that the polysaccharide hydrolysates were linear glucan containing 1, 4-glycosidic bonds. And gluco-oligosaccharides with the degrees of polymerization (DP) of 4-11 were obtained after partial acid hydrolysis.
ABSTRACT
Objective In order to analyze of poisoning causes,a new method was established utilizing hydrophilic liquid chromatography-tandem triple quadrupole mass spectrometer (HILIC-MS/MS) coupled with dispersive solid phase extraction for rapid qualitative and quantitative analysis of tetrodotoxin in nassarius and shellfish.Methods Sample (1.0 g) was extracted with 0.1％ acetic acid in boiling water bath,purified by dispersive solid phase extraction with 50 mg hydrophilic-lipophilic balance (HLB),5 mg graphitized carbon black (GCB) and protein precipitation with acetonitrile,and then filtered through a polytetrafluoroethylene (PTFE) membrane.The analytes were separated on a HILIC column,and detected in selected reaction monitoring (SRM) mode via positive electrospray ionization.The matrix matching and external standard method was used for quantification.Results Tetrodotoxin showed good linearity in the concentration range between 2.0 and 40.0 ng/ml,the correlation coefficient was higher than 0.999.The detection limit of tetrodotoxin in seafood was 10.0 pg/kg.The rates of recovery varied between 74.2％ and 87.9％ with relative standard deviations from 2.3％ to 9.1％ at spiked concentrations of 25,100 and 200 pg/kg.The proposed method was applied in the detection of tetrodotoxin in shellfish and nassarius from coastal cities of Zhejiang Province.Conclusion The method was accurate,fast,easy to operate,which could meet the requirements of public health emergency testing or routine testing.