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Polysaccharides and free monosaccharides are important active components in Cistanches Herba, which have functions of anti-aging and immunological activity regulation. The study of monosaccharide composition in polysaccharide and free monosaccharide can lay a foundation for the study of primary structure, spatial structure of Cistanche polysaccharide and biological activity of Cistanches Herba. In this study, a method of water extraction and alcohol precipitation was used to extract Cistanche polysaccharide. Trifluoroacetic acid was selected as the hydrolytic acid for polysaccharide hydrolysis. An orthogonal experimental method is established. Three levels of acid concentration, hydrolysis temperature and hydrolysis time were selected to investigate the optimal hydrolysis condition. The optimal hydrolysis condition was 0.08 mol·L-1 trifluoroacetic acid hydrolysis at 100 ℃ for 3 h. The free monosaccharides of Cistanches Herba were extracted by water extraction. The established ion chromatogram integrated pulsed amperometry method can efficiently separate 11 monosaccharides in a short time. The method has good repeatability and high sensitivity, methodological experiment results meet the requirements of quantitative determination. It can accurately determine the monosaccharide composition of Cistanche polysaccharide and free monosaccharide content. Ion chromatography does not require derivatization operation and the pre-treatment steps are simple. This method can measure fructose, but PMP derivation-HPLC method can't. The monosaccharide composition of Cistanche polysaccharide include fucose, arabinose, rhamnose-galactose, glucose, xylose, mannose, fructose, ribose and glucuronic acid, among which the contents of glucose and fructose are relatively high. The free monosaccharides in the water extract of Cistanches Herba include glucose, fructose and mannose.
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The paper aims to establish the method to determine the monosaccharide composition and monosaccharide ratio in propylene glycol alginate sodium sulphate (PSS). Samples were hydrolyzed with trifluoroacetic acid, neutralized with sodium hydroxide solution after the reaction conditions for sample pretreatment were optimized via orthogonal analysis. A high performance anion exchange chromatograghy (HPAEC) coupled with pulsed amperometric detector (PAD) was performed on a CarboPac®PA20, using 200 mmol·L-1 sodium hydroxide solution and 1 mol·L-1 sodium acetate solution as mobile phase. The established HPAEC-PAD method was validated by testing the linear relationship, precision and accuracy, and showed exclusive, sensitive, rapid and wide use. The monosaccharide composition of PSS from different manufacture can be accurately determined with great significance for the structural identification of PSS.
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OBJECTIVE： To establish pre- column derivatization-HPLC fingerprint of polysaccharide from Achyranthes bidentata，and to determine the contents of monosaccharide components ，so as to provide reference for quality evaluation of A. bidentata decoction pieces. METHODS ：Taking 10 batches of A. bidentata decoction pieces from different manufacturers as samples，the polysaccharides were extracted by water extraction and alcohol precipitation ，hydrolyzed by trifluoroacetic acid ，and then derivatized by 1-phenyl-3-methyl-5-pyrazolone for HPLC analysis. The determination was performed on Hanbon Hedera C 18 column with column temperature of 30 ℃ at the flow rate of 1.2 mL/min. The mobile phase consisted of acetonitrile- 0.02 mol/L ammonium acetate solution （gradient elution ）. The detection wavelength was set at 250 nm，and sample size was 20 μL. HPLC fingerprint was established and similarity evaluation was performed for 10 batches of A. bidentata polysaccharide by using TCM Chromatogramic Fingerprint Similarity Evaluation System （2012A edition ）. The common peak was identified by comparing with the reference substance ，and cluster analysis was performed by using SPSS 25.0 software. The contents of identified monosaccharides were determined by HPLC. RESULTS ：The similarity of 10 batches of A. bidentata polysaccharide were all higher than 0.95. Nine common peaks were fixed and a total of 5 common peaks were identified ，namely anhydrous glucose （peak 1）， mannose（peak 2），rhamnose（peak 4），galacturonic acid （peak 5）and arabinose （peak 7）. Results of cluster analysis showed that S1 sample was classified into one category ；S2，S5，S8 and S 9 samples were clustered into one category ；S3，S4，S6，S7 and S10 samples were clustered into one category. Results of content determination showed that the contents of anhydrous glucose in 10 batches of samples ranged from 6.17 to 17.55 mg/g；those of mannose ranged from 3.31 to 7.66 mg/g；those of rhamnose ranged from 38.80 to 73.97 mg/g；those of galacturonic acid ranged from 2.49 to 8.95 mg/g；those of arabinose ranged from 11.30 to 28.58 mg/g. CONCLUSIONS ：Established pre-column derivatization HPLC fingerprints and content determination method can provide reference for quality evaluation of A. bidentata decoction pieces.
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Objective To analyze and identify the chemical components in the Nocardia rubra cell wall skeleton (Nr-CWS), and to determine the contents of monosaccharides accurately. Methods The extract of Nr-CWS was separated and analyzed by UHPLC-Q-TOF/MS method. The chemical components were quickly identified by matching the data with the information in the Metlin database. The monosaccharide contents in the Nr-CWS extract were determined by UHPLC-MS/MS method after derivatization. Results A total of 64 chemical components were identified in the extract of Nr-CWS, including amino acids, monosaccharides and so on. A assay method for 8 monosaccharides by UHPLC-MS/MS was successfully established. The content of arabinose in Nr-CWS was the highest, followed by galactose, which indicated that the main polysaccharide components in Nr-CWS may be composed of these monosaccharides. Conclusion In this study, we analyzed the main chemical components of Nr-CWS, which are amino acids, fatty acids and so on. The content of monosaccharide after polysaccharide hydrolysis was determined by UHPLC-MS/MS. This will lay a foundation for the screening of the active components of Nr-CWS and the study of its pharmacological mechanism.
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OBJECTIVE：To establish pre -column derivatization-HPLC fingerprint of Polyporus polysaccharide ，and to determine the contents of main monosaccharide components ，so as to provide reference for quality evaluation of Polyporus umbellatus. METHODS ：Polyporus polysaccharide was extracted with boiling water and precipitated by ethanol and deproteinized by Sevage from 11 batches of P. umbellatus from different producing areas. The samples were firstly hydrolyzed with trifluoro-acetic acid （TFA）and then derivatized by 1-phenyl-3-methyl-5-pyrazolone（PMP）. HPLC analysis was then conducted. The determination was carried out on HypersiL BDS C 18 column with mobile phase composed of 0.1 mol/L phosphate buffer （pH 6.84）-acetonitrile（84∶16，V/V）by gradient elution at the flow rate of 1.0 mL/min. The detection wavelength was set at 254 nm， and column temperature was 30 ℃. The sample size was 20 µL. The similarity of 11 batches of Polyporus polysaccharide was evaluated by using TCM Chromatographic Fingerprint Similarity Evaluation System （2012A edition ），and the contents of main monosassharide components were detected. The peak was identified by comparing with the reference substance ，and cluster analysis was performed by using SPSS 23.0 software. RESULTS ：In HPLC fingerprints of the 11 batches of samples ，3 common peaks were identified ，namely mannose ，glucose and galactose. The similarity of all samples was above 0.94. Cluster analysis classified 11 batches of samples into three categories. S 1-S6，and S 8 were grouped into category 1；S7，S10 and S 11 were grouped into category 2；S9 was individually grouped into one category. Results of content determination showed that the contents of mannose ranged from 1.571 to 8.771 mg/g；those of glucose ranged from 26.072 to 132.194 mg/g，and those of galactose ranged from 3.420 to 36.593 mg/g. CONCLUSIONS ：Established pre-column derivatization HPLC fingerprints can provide reference for quality evaluation of P. umbellatus . The monosaccharide composition of different batches of Polyporus polysaccharide is the same ；there is no significant correlation between fingerprint characteristic peak and the origin of herbs ；there is significant difference in the content of monosaccharide of P. umbellatus .
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Objective To establish a method for analyzing total polysaccharide content and its monosaccharide composition in the caulis polygoni multiflori mixture. Methods The polysaccharides of the caulis polygoni multiflori mixture were extracted by water-extraction and alcohol-precipitation. After the treatment with phenol-sulfuric acid, the content of total polysaccharides in the preparation was determined by ultraviolet spectrophotometry. In addition, after the polysaccharide was hydrolyzed into monosaccharides wtih trifluoroacetic acid, the hyrolysate was derivatized with PMP, and then the PMP derivates of monosaccharides were analyzed by HPLC method. Yilite krosmasil C18(4.6 mm×250 mm, 5 μm) was used at 30 ℃. Acetonitrile-0.1% NaH2PO4 (pH=6.8) (16:84) was the moblie phase with a flow rate of 1.0 ml/min. The detecting wavelength was at 250 nm. The injection volume was 20 μl. Results concentration of D-anhydrous glucose in the range of 21 ~ 105 μg/ml had a good linear relationship with the absorbance. The linear regression equation was A= 0.007x+0.0105, r=0.9982. The average recovery rate was (100.45±1.57)% (n=6). The average contents of total polysaccharides in four batches of samples were 14.24, 21.09, 17.85 and 18.17 mg/ml. The polysaccharide of the caulis polygoni multiflori mixture mainly consisted of D-mannose, D-glucosamine D-hydrochloride, D-Galacturonic acid, D-glucose, galactose, L-arabinose. The monosaccharides peak area ratios were about 9.10:0.26:1.00:3.02:4.14:2.12. Conclusion The method is accurate and reliable, and can be used for the determination of total content of polysaccharides and the analysis of monosaccharide composition in the preparation.
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OBJECTIVE：To estab lish the fingerprint ，analyze the monosaccharide composition and content ，investigate the inhibitory effects of the polysaccharide from Desmodium styracifolium on α-glucosidase in vitro . METHODS ：Polysaccharide from D. styracifolium was prepared by water extraction and ethanol precipitation. After hydrolyzed by TFA and derived by PMP ，HPLC method was adopted to establish the fingerprint （using glucose peak as reference ），and analyze the constituent and content of monosaccharide. The content determination was performed on Phenomenex Luna C 18 column with mobile phase consisted of acetonitrile-0.05 mol/L potassium phosphate （pH adjusted to 6.8 with sodium hydroxide ）in gradient elution at the flow rate of 0.8 mL/min. The detection wavelength was set at 250 nm，and column temperature was set at 30 ℃. The sample size was 10 μL. Using acarbose as control ，PNPG assay was used to investigate the α-glucosidase inhibitory activity of polysaccharide from D. styracifolium. RESULTS ：There were 9 common peaks in HPLC fingerprints of 18 batches of samples ，and the similarity of 15 batches of samples was higher than 0.90. Totally 7 peaks were identified as mannose ，rhamnose，galacturonic acid ，glucose， galactose，xylose and arabinose. The contents of rhamnose ，galacturonic acid ，glucose，galactose and arabinose were 0.471-2.092， 1.379-8.919，2.560-35.679，1.194-6.905，0.566-4.158 mg/g，respectively. Based on rhamnose ，the molar ratios of the other four monosaccharides were 1.58-4.07，2.26-19.95，2.20-4.21 and 1.31-2.86，respectively. The inhibitory activity of polysaccharide from D. styracifolium on α-glucosidase increased with the increase of dose ，and the half inhibitory concentrations of it was 0.70 mg/mL， lower than 7.76 mg/mL of acarbose （positive control ）. CONCLUSIONS ：Glucose is the main component of D. styracifolium polysaccharide in different batches ，and the contents of monosaccharides are different. The polysaccharide from D. styracifolium have significant inhibitory activity on α-glucosidase，which is better than that of acarbose.
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Characterization of the polysaccharides and monosaccharides of Bupleurum chinense was undertaken to identify differences in the Bupleurum chinense's sugar profiles, so as to provide a basis for the identification of different varieties. High performance liquid chromatography (HPLC) was used to generate chromatograms of the total polysaccharides of Bupleurum using an Evaporation Light Detector (ELSD), and a monosaccharide chromatogram was generated using a UV-detector (UV) following polysaccharide derivatization. The data were analyzed using SIMCA software and SPSS software to distinguish different varieties of Bupleurum. The results show that the yield of polysaccharides from Bupleurum falcatum is the highest, while the yield of polysaccharides from Bupleurum chinense is the lowest. The polysaccharide spectrum shows that the molecular weights of the polysaccharides in different Bupleurum differ, and their percentages of the total peak area are also different. The four Bupleurum polysaccharides are composed of mannose, glucuronic acid, rhamnose, galacturonic acid, glucose, galactose, and arabinose, but differ in length. The ratio of glucose to arabinose in Bupleurum chinense, Bupleurum scorzonerifolium, Bupleurum falcatum and Bupleurum marginatum var. stenophyllum is: 3.0-4.0, 5.5-7.0, 12.0-17.0, 9.0-12.0. In this study, a sugar profile technique was developed to provide a new method for the identification of different varieties of Bupleurum.
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Objective:To characterize the structure of polysaccharide isolated from Linggui Zhugan Tang（LGZGT）,including monosaccharide composition and functional group detection, investigate the difference of the antioxidant activities of crude polysaccharide（CP） and pure polysaccharide（PP）, and provide the basis for the quality evaluation of LGZGT by in vitro bioassay. Method:The average molecular weight of CP was analyzed by high performance gel chromatography（HPGPC）. Gas chromatography-mass spectrometry（GC-MS） and fourier transform infrared（FT-IR） were employed to determine the structure of the polysaccharide. The antioxidant activities of CP and PP samples were evaluated on the basis of 1-diphenyl-2-picrylhydrazyl（DPPH） radical scavenging activity and OH radical scavenging activity. Result:The total polysaccharide was composed of single peaks, with a molecular weight of 3 689 Da. It was mainly composed of arabinose, mannose, glucose, galactose and fructose with a molar ratio of 6.85∶1.00∶109.21∶1.04∶21.82. Among them，glucose and fructose were the predominant components. In addition， IR study indicated the presence of pyranose and anomeric configurations in glycan structure, with two stereoisomers of glycosidic bond (α-glycosidic bond and β-glycosidic bond). It was found that the total polysaccharide had the ability of scavenging DPPH and hydroxyl radicals, and the activity of crude polysaccharide was better than that of refined polysaccharide. It was found in antioxidant research that the total polysaccharide had the ability of scavenging DPPH and hydroxyl radicals, and the activity of CP was better than that of PP. Furthermore, LC-Q-TOF-MS was used to qualitatively analyze the other components in CP, which indicated that it was related to the adsorption of pentacyclic triterpenoids in Glycyrrhiza uralensis. Conclusion:The polysaccharides and pentacyclic triterpenoids in LGZGT are the material basis for the antioxidative effect of LGZGT. The antioxidative activity determined by in vitro bioassay can be used as an evaluation index for the overall quality control of LGZGT.
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The non-starch polysaccharides,mainly composed of glucomannans,are the major bioactive compounds in Dendrobium catenatum. In order to evaluate the quality of the medicinal materials and guide the production and processing,a quantification method of non-starch polysaccharides was established by stems of D. catenatum C15 strain collected from the pear epiphytic cultivation. The non-starch polysaccharides were obtained by " water extraction,α-amylase pretreatment,and alcohol precipitation once" method. The contents of starches,non-starch polysaccharides and monosaccharides were analyzed. In addition,the system suitability was tested. Compared with method of the Chinese Pharmacopoeia( 2015 edition),the contents of total polysaccharides,glucose,and mannose were decreased by 20. 9%,58. 8% and 1. 6% respectively. The method effectively digested starch and retained non-starch polysaccharides,and the analysis result was accurate and repeatable. Therefore,it is suitable for the content measurement of non-starch polysaccharides of D. catenatum. Furthermore,it could be an alternative method for quality control of D. catenatum and a reference in the determination of non-starch polysaccharides in other starch-containing medicinal materials.
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Dendrobium , Chemistry , Monosaccharides , Phytochemicals , Polysaccharides , StarchABSTRACT
In order to provide scientific basics for exploitation and sufficient application of Polyporus umbellatus resources and study the monosaccharide composition of P. umbellatus polysaccharides,the anthrone-sulfuric acid method was applied to compare polysaccharide content of P. umbellatus from 17 producing areas. The monosaccharides were derived by 1-phenyl-3-methyl-5-pyrazolone( PMP) and the derivatives were identified by UPLC-MS/MS and the content of each monosaccharide component was determined simultaneously. The results demonstrated that there was a certain difference in total polysaccharide content of P. umbellatus from different regions,and the content of total P. umbellatus polysaccharide from Shaanxi province and Sichuan province( 1. 15% and 1. 90%) was relatively higher than that of others areas. Polysaccharides from P. umbellatus was mainly composed of eight monosaccharides,including glucose,glucuronic acid,galactose,ribose,xylose,arabinose,mannose and fucose. The contents of glucose( 17. 65 mg·g-1) was higher than others. The ribose was the lowest( 0. 13 mg·g-1). In addition,fructose,rhamnose and galacturonic acid were also detected in some samples. Furthermore,the results of cluster analysis( CA) and principal component analysis( PCA) indicated that totally 17 batches of P. umbellatus polysaccharide could be classified into three clusters,samples collected from Wuchang in Heilongjiang province were clustered into one group separately. The study can provide a basis for rational utilization of P. umbellatus resources,and also implies the sequence of monosaccharide linking and pharmacological activity of P. umbellatus polysaccharides.
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China , Chromatography, High Pressure Liquid , Geography , Monosaccharides , Chemistry , Polyporus , Chemistry , Polysaccharides , Chemistry , Tandem Mass SpectrometryABSTRACT
Objective To determine the monosaccharide composition of polysaccharides in different Cordyceps powder preparations by capillary zone electrophoresis (CZE). Methods The orthogonal experiment was used to optimize the total degradation conditions of the extracted polysaccharides; The monosaccharide composition was determined by CZE method using 1-phenyl-3-methyl-5-pyrazolone (PMP) as a derivatizing reagent. The electrophoresis conditions were as follows: uncoated fused silica capillary column (70 cm × 50 μm i.d., effective length was 61 cm); 40 mmol/L borax solution (pH 10.1) was used as the running buffer; The detection wavelength was set at 245 nm; The operation voltage was 13 kV; Hydrodynamic pressure injection was employed (10 cm × 8 s). Results The optimum hydrolysis conditions were as follows: 1.0 mol/L sulfuric acid solution, hydrolysis temperature 100 ℃, and hydrolysis time 5 h. The common monosaccharides of polysaccharides in Xinganbao Capsule, Bailing Capsule, Zhiling Junsi Capsule, and Jinshuibao Capsule were xylose, glucose, mannose, and galactose, but the content was different. The monosaccharide components contained in the polysaccharides of the four preparations were basically the same as those of the Cordyceps sinensis, and monosaccharides included xylose, rhamnose and glucuronic acid which were not reported in Cordyceps sinensis were detected. Each monosaccharide had a good linear relationship with the concentration range of 0.01 to 0.20 mg/mL, the limits of detection (LOD, S/N = 3) and the limits of quantification (LOQ, S/N = 10) ranged from 0.205 to 0.397 μg/mL and 0.684 to 1.323 μg/mL, respectively. RSDs of the precision test were 0.8%-3.6%, RSDs of the repeatability test were 2.7%-4.7%, and 2.8%-4.7% in the stability test. The recovery rate of the method showed that the mean recoveries of glucose and galactose ranged from 96.1% to 99.8% and 95.1% to 99.6% respectively, and RSD values fell within 1.0%-2.0% and 1.5%-1.9%, respectively. Conclusion The method is fast and efficient, and provides a reference for the improvement of quality standard of Cordyceps powder preparations and the intensive study on the substitution of Cordyceps powder for Cordyceps sinensis.
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Objective: To study the relative molecular mass and composition of Ligustrum lucidum polysaccharide purified by dialysis, and provide the theories for the relationship between the biological activity and the internal composition. Methods: In this paper, L. lucidum as an object was studied. Polysaccharide was isolated by water extraction and ethyl alcohol precipitation and purified by Sevage method, hydrogen peroxide and dialysis. After the acidolysis of trifluoroacetic acid, the molecular weight was measured by GPC, and the composition of polysaccharide was analyzed by HPLC-RID. Results: The Mw of polysaccharide was 10 721, and the Mn of polysaccharide was 10 673. L. lucidum polysaccharide consisted of glucose, rhamnose, and arabinose, the molar ratio of these monosaccharide was 9.148.105.18. Conclusion: The purified polysaccharide composition is more homogeneous, and the monosaccharides of polysaccharides was easily analyzed by HPLC-RID without column derivatization.
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Objective: To extract, separate and purify homogeneous polysaccharides from Rubus sachalinensis and study on monosaccharide component and immunomodulatory activity. Methods: The crude polysaccharides of Rubus sachalinensis (RSP) were extracted by hot water. The Cellulose DE-52 and Sephadex G-100 columns were used to separate and purify homogeneous polysaccharides. The relative molecular mass was analyzed by high-performance gel permeation chromatography, and the monosaccharide composition and structure were preliminarily identified by GC, IR and NMR. The effects on proliferation function of mice spleen lymphocyte proliferation were determined by CCK-8, and the effects on the release capacity of IL-2, IFN-γ and TNF-α were determined by the ELISA kit. Results: Two homogeneous polysaccharides, RSP1-1 and RSP1-2, were separated and purified, with molecular weights of 13 227 and 9 343 determined by HPGPC. They mainly contained arabinose, mannose, glucose and galactose, with the mole ratios at 9.5:7.0:10.3:18.6 and 5.7:11.1:10.3:14.2, respectively. The structure of RSP1-1and RSP1-2 was analyzed by IR and NMR, and RSP1-1 might mainly contain α-1,3-Ara, β-1,4Gal, β-1,6-Glc, β-1,3-Man, and RSP1-2 might mainly contain β-1,4-Gal, β-1,6-Glc, β-1,3-Man. At 5-200 μg/mL, RSP1-1and RSP1-2 stimulated proliferation of spleen lymphocytes (P < 0.05) and promoted lymphocytes to secrete IFN-γ and TNF-α. At 5 μg/mL, RSP1-1and RSP1-2 promoted lymphocytes to secrete IL-2. Conclusion: RSP1-1and RSP1-2 are natural homogeneous polysaccharides that are obtained from this plants for the first time. Its purity and structure were further characterized by IR and NMR. These two homogeneous polysaccharides promoted the proliferation of splenic lymphocyte in different degrees and promoted lymphocytes to secrete IL-2, IFN-γ and TNF-α that all possessing immunomodulating activity.
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Objective To analyze the monosaccharide composition and the relative content of different alcohol precipitations of Panax japonicus polysaccharides.Methods The four components of Panax japonicus polysaccharides were isolated by stepwise ethanol precipitation method.The four components of Panax japonicus polysaccharides were hydrolyzed with trifluoroacetic acid (TFA) and derivated by l-phenyl-3-methyl-5-pyrazolo (PMP),respectively.The monosaccharide composition and relative content were analyzed using LC-FT-ICR-MS and HPLC-UV method.Results The four components of Panax japonicus polysaccharides consisted of glucose,rhamnose,galacturonic acid,galactose,arabinose,and glucose was the main monosaccharide.With the increase of ethanol concentration,the relative content of Ara increased gradually,as while the GalA decreased.Conclusions Precolumn derivation HPLC method was successfully applied for the determination of the monosaccharides in Panax japonicus polysaccharides,and there were differences in the four ethanol precipitations of Panax japonicus polysaccharides.The study can provide a basis for the separation of Panax japonicus polysaccharides.
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The flower of Hibiscus syriacus has good ornamental and edible-medicinal values.In this study,four samples of two varieties,namely white multiple petals flower and pink multiple petals flower,were selected as test materials.And the optimum extraction conditions,relative molecular weight,monosaccharide composition and antioxidant activity of polysaccharides in flower were investigated.Through single factor experiment and response surface,the optimal extract conditions of polysaccharide were designed as follows:extraction temperature at 96.8℃,ratio of material to liquid of 43.5∶1 m L·g~(-1),extraction time of 3.1 h.Polysaccharides of H.syriacus flowers were analyzed by high performance gel chromatography.The average molecular masses of the 4 polysaccharide samples were1.49×10~5,1.25×10~5,1.01×10~5,1.37×10~5,respectively.Polysaccharides of H.syriacus flowers were mainly composed of glucose,mannose,galactose,rhamnose and arabinose by pre-column derivatization HPLC.The ratio of galactose was the highest in five monosaccharide,and the ratio of galactose to glucose was 1.656-4.496.In addition,crude polysaccharides of H.syriacus flowers showed potential antioxidant activity by 1,1-diphenyl-2-picrylhydrazyl radical(DPPH)assay,total reducing capacity assay and ABTS assay in vitro,and its antioxidant effect showed a good dose-effect relationship with the concentration of crude polysaccharides.Among the tested varieties,polysaccharides of pink multiple petals flower and white multiple petals flower had the same molecular masses and monosaccharides composition,but the antioxidant activity of the polysaccharides of pink multiple petals flower was higher than that of the white flowers.The results of this study can provide a theoretical basis for the application of H.syriacus flower in the field of functional foods.
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Antioxidants , Chemistry , Pharmacology , Flowers , Chemistry , Hibiscus , Chemistry , Monosaccharides , Chemistry , Phytochemicals , Chemistry , Pharmacology , Pigmentation , Polysaccharides , Chemistry , PharmacologyABSTRACT
Abstract Agaricus subrufescens is a basidiomycete which is studied because of its medicinal and gastronomic importance; however, less attention has been paid to its preservation. This study aimed to evaluate the effect of sucrose addition to substrate and cryotube on the viability of Agaricus subrufescens cryopreserved at -20 °C and at -75 °C for one and two years. Zero, 10% or 20% sucrose was added to potato dextrose agar or wheat grain. The mycelia were cryopreserved in the absence of cryoprotectant or with sucrose solutions at 15%, 30% or 45%. After one or two years at -75 °C or at -20 °C, mycelia were thawed and evaluated about viability, initial time of growth, colony diameter and genomic stability. Cryopreservation at -20 °C is not effective to keep mycelial viability of this fungus. Cryopreservation at -75 °C is effective when sucrose is used in substrates and/or cryotubes. Without sucrose, cryopreservation at -75 °C is effective only when wheat grains are used. Physiological characteristic as mycelial colony diameter is negatively affected when potato dextrose agar is used and unaffected when wheat grain is used after two-year cryopreservation at -75 °C. The fungus genome does not show alteration after two-year cryopreservation at -75 °C.
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Agaricus/growth & development , Cryopreservation/methods , Cryoprotective Agents/metabolism , Freezing , Seeds/microbiology , Sucrose/metabolism , Triticum/microbiology , Agaricus/radiation effects , Genomic Instability/radiation effects , Microbial Viability/radiation effects , Mycelium/growth & development , Mycelium/radiation effects , Time FactorsABSTRACT
This study is to establish a pre-column derivatization procedure with 1-phenyl-3-methyl-5-pyrazolone (PMP) UPLC-MS/MS method for the determination of the monosaccharide composition of 12 polysaccharides. At the same time, the monosaccharide components of polysaccharides in Armillaria gallica were analyzed. The separation was performed on a ACQUITY ZORBAX RRHD Eclipse Plus C₁₈ column(2.1 mm×100 mm, 1.8 μm),using 95% acetonitrile (A) and ammonium acetate-5% acetonitrile-water (B) as mobile phase with gradient elution. The target components were detected in multiple-reaction monitoring (MRM) mode by mass spectrometry with electrospray ionization (ESI) source operated in ionization mode. The results showed that based on the monosaccharides detection method established by UPLC-MS/MS, the linearity of the 12 monosaccharides components were linear in their linear range (R²>0.990), and the recovery rate were 92.30%-105.6%. 11 monosaccharides such as fructose, mannose, and glucose were detected in A. gallica samples. The method established in this experiment is robust, highly reproducible and accurate, and is suitable for the determination of monosaccharide components such as A. gallica.
Subject(s)
Chromatography, High Pressure Liquid , Chromatography, Liquid , Monosaccharides , Polysaccharides , Tandem Mass SpectrometryABSTRACT
Obejctive:To study the extraction, separation, physical and chemical properties and antioxidant activity of the crude polysaccharide from Alhagi sparsifolia Shap.stem-branch.Methods: The crude polysaccharide from Alhagi sparsifolia Shap.stem-branch was extracted by ethanol subsiding method .The total sugar content was determined by phenol-sulfuric acid method and the pro-tein content was determined by coomassie brilliant blue method .The content of uronic acid was determined by carbazole sulfuric acid method and the monosaccharide composition and the relative molar ratio were determined by GC .The antioxidant activities in vitro were evaluated by determining the reducing power of polysaccharide from Alhagi sparsifolia Shap stem-branch and its removal ability to 1,1-diphenyl-2-trinitrophenylhydrazine ( DPPH) radicals and hydroxyl radicals .Results: The total sugar content of crude polysaccharide from Alhagi sparsifolia Shap.stem-branch was 73.2％, and uronic acid accounted for 27.2％of the total sugar content of crude poly-saccharide.The content of protein was 17.6％.The crude polysaccharide from Alhagi sparsifolia Shap .stem-branch was composed of Rha, Ara, Xyl, Man, Glc, Gal, GlcA and GalA, and the relative molar ratio was 1.05:1.00:1.25:0.52:3.05:1.31:0.47:4.78. The reducing power of the polysaccharide from Alhagi sparsifolia shap.stem-branch and the clearance rate on DPPH radicals and hy-droxyl radicals increased with the increase of polysaccharide concentration .Conclusion:The crude polysaccharide from Alhagi sparsi-folia Shap.stem-branch was extracted , and the physical and chemical properties and antioxidant activity in vitro were studied, which provide foundation for the further investigation and comprehensive utilization of Alhagis parsifolia Shap.stem-branch.
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Objective: To determine the relative molecular weight (Mw), monosaccharide composition, and structures of polysaccharides from the leaves of Eriobotrya japonica. Methods: The polysaccharides (PEL60) were extracted from the leaves of E. japonica with hot water, followed by precipitating with ethanol, freezing, and drying. PEL60-A was purified by DEAE-Sepharose Fast Flow ion column and Sephacryl S 500 High-Resolution gel column chromatography from PEL60 of the leaves of E. japonica. The purity and molecular weight of PEL60-A was detected by high performance liquid chromatography (HPLC), and the monosaccharide composition and molecular ratio of the PEL60-A was analyzed by gas chromatography-mass spectrometry (GC-MS). Results: The PEL60-A, with Mw of 2.69 × 105. The composition of monosaccharide was mainly L-rhamnose and L-fucose and a small amount of D-xylose, D-mannose, D-glucose, and D-galactose with the molar ratio of 0.781.000.150.090.110.19. Conclusion: The study on isolation, purification, and preliminary identification of monosaccharide composition from leaves of E. japonica provided the important scientific theoretical basis for the fine processing of the leaves of E. japonica.