ABSTRACT
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.
Subject(s)
China , Chromatography, High Pressure Liquid , Geography , Monosaccharides , Chemistry , Polyporus , Chemistry , Polysaccharides , Chemistry , Tandem Mass SpectrometryABSTRACT
OBJECTIVE:To establish the method for the content determination of L-Hyp and collagen in gelatinous medicinal material,and to compare the contents of two components in reference medicinal material and commercially available medicinal material.METHODS:Pre-column derivatization was adopted for pretreatment.The content of L-Hyp was determined by HPLC.The determination was performed on Kromasil C18 column with mobile phase consisted of acetonitrile-0.1 mol/L sodium acetate buffer (pH 6.5,7 ∶ 93,V/V)-acetonitrile-water (4 ∶ 1,V/V) (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set at 254 urn,and column temperature was set at 43 ℃.The sample size was 20 μL.The content of collagen was calculated by using convert coefficient.RESULTS:The linear range of L-Hyp were 2.5-40 μg/mL(r=0.999 9).LOQ was 0.20 μg/mL,and LOD was 0.05 t g/mL.RSDs of precision,stability and reproducibility tests were all lower than 4.0％.The recoveries were 96.03％-102.07％ (RSD =2.20％,n =9).There was difference in the contents of L-Hyp and collagen among 28 batches of commercially available medicinal material.The contents of two components in 13 batches of commercially available Colla Corii Asini were relative close to reference medicinal material;5 batches of Colla Carapacis et Plastri Testudinis and 7 batches of Cervi Comus Colla were much higher than those of reference medicinal material.CONCLUSIONS:The method is accurate,reliable and suitable for the content determination of L-Hyp and collagen in gelatinous medicinal material.
ABSTRACT
OBJECTIVE:To establish the method for the content determination of L-Hyp and collagen in gelatinous medicinal material,and to compare the contents of two components in reference medicinal material and commercially available medicinal material.METHODS:Pre-column derivatization was adopted for pretreatment.The content of L-Hyp was determined by HPLC.The determination was performed on Kromasil C18 column with mobile phase consisted of acetonitrile-0.1 mol/L sodium acetate buffer (pH 6.5,7 ∶ 93,V/V)-acetonitrile-water (4 ∶ 1,V/V) (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set at 254 urn,and column temperature was set at 43 ℃.The sample size was 20 μL.The content of collagen was calculated by using convert coefficient.RESULTS:The linear range of L-Hyp were 2.5-40 μg/mL(r=0.999 9).LOQ was 0.20 μg/mL,and LOD was 0.05 t g/mL.RSDs of precision,stability and reproducibility tests were all lower than 4.0％.The recoveries were 96.03％-102.07％ (RSD =2.20％,n =9).There was difference in the contents of L-Hyp and collagen among 28 batches of commercially available medicinal material.The contents of two components in 13 batches of commercially available Colla Corii Asini were relative close to reference medicinal material;5 batches of Colla Carapacis et Plastri Testudinis and 7 batches of Cervi Comus Colla were much higher than those of reference medicinal material.CONCLUSIONS:The method is accurate,reliable and suitable for the content determination of L-Hyp and collagen in gelatinous medicinal material.
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Objective To establish a pre-column derivation HPLC-UV method for the content determination of hyodeoxycholic acid in artificial calculus bovis. Methods The hyodeoxycholic acid was derived by 2-bromo-2’-acetonaphthoneat using triethylamine as the catalyst in 60 ℃ water bath. After that, a HPLC-UV method was established to determine the content of hyodeoxycholic acid in artificial calculus bovis. Results When the derivatising time at 60 ℃ water bath was 50 min, the radio of the molar amount of derived reagents and hyodeoxycholic was over 20:1 and the radio of catalyst and hyodeoxycholic was over 15:1; the reaction was completed. The calibration curve was linear within the range of 0.1–2 μg for hyodeoxycholic acid (r=0.999 7), and the average recovery was 97.85% (RSD=1.6%). In this sample, the content of hyodeoxycholic is 4.12%. Conclusion The method is with high sensitivity, highly reproducible, reliable and accurate for the content determination of hyodeoxycholic acid in artificial calculus bovis.
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Objective:To establish a method for the separation and determination of ketoprofen enantiomer .Methods:A pre-col-umn derivation RP-HPLC method was used with L-alanine-β-naphthylamine ( L-Ala-β-NA) as the derivation reagent .The RP-HPLC conditions were as follows: a Hypersil ODS-2 column (250 mm ×4.6 mm,5 μm) was applied, the mobile phase was acetonitrile-0.025 mol· L-1 phosphate buffer solution (40∶60, v/v) and the flow rate was 1.0 ml· min-1 , the detection wavelength was set at 245 nm and the column temperature was 25℃.The injection volume was 10μl.Results:Base line separation was achieved for the sep-aration of enantiomer from ketoprofen , and the retention time for S-(+) -ketoprofen and the R-(-) -ketoprofen was 24.2 min and 26.0 min, respectively.Dexketoprofen within the range of 0.025-0.125 mg had a good linear relationship (r=0.998 1) and the aver-age recovery was 90.93%(RSD =4.10%, n=9 ).Conclusion:The method is simple, accurate and reliable, which can be applied in the separation and determination of ketoprofen .
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Objective:To establish a RP-HPLC method with pre-column derivation for determining the content of stilbene glyco-sides in Yening granules. Methods:DNS-Cl was used as the derivation reagent, a Wondasil-C8 column (250 mm × 4. 6 mm, 5 μm) was used as the stationary phase and 30mM ammonium phosphatere gulation (pH 3. 5)-acetonitrile (78∶22) was applied as the mobile phase, a fluorescence detector (λex=370 nm,λem=560 nm) was used, the flow rate was 1. 0 ml·ml-1 , the column temperature was 30℃ and the injection volume was 20 μl. Results: There was no interference from the other substances in the preparation. The linear range of the two stilbene glycosides was 0.071-27.280 mg·ml-1(r=0.999 9). The average recovery was 99.04%(RSD=0. 1%, n= 9). Conclusion:The method is simple, quick, sensitive and accurate, which can be used for the quality control of Ye-ning granules.
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Objective To study the metabolism of amino acids in AIDS patients for the purpose of providing appropriate nutritional support.Method High performance liquid chromatography combined with pre-column derivatization technology was used to detect free amino acids in plasma from 22 untreated AIDS patients,16 HAART treated AIDS patients and 30 healthy controls.Statistic analysis was performed through SPSS software.Results The concentration of alanine(Ala) was significantly increased and the concentration of phenoalanine(Phe) was remarkably decreased in AIDS patients as compared with the healthy controls(P