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Objective To establish a method for content determination of total flavones and polysaccharides in Fangshu Qingre mixture by UV-Vis Spectrophotometry. Methods The contents of total flavones and polysaccharides in Fangshu Qingre mixture were determined by UV-Vis spectroscopy with rutin and anhydrous glucose as reference substance, and the wavelength was set at 508 nm and 487 nm. Results The contents were from 0.00 to 59.20 μg/ml for total flavones and from 10.92 to 109.20 μg/ml for total polysaccharides in Fangshu Qingre mixture. The recoveries of total flavones and total polysaccharides were 104.4% and 104.8% respectively. Conclusion The method of using ultraviolet spectroscopy was simple, reproducible, accurate and reliable, which could be preferably used as the method for content determination of total flavones and polysaccharides in Fangshu Qingre mixture.
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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 establish a method for determining the content of total polysaccharides in decoction pieces of Polyporus,analyze the content of total polysaccharides in samples with different sources and grades. Method: The relative molecular weight and the polydispersity index of polysaccharides in decoction pieces of Polyporus were measured by a high performance gel chromatography coupled with a multi-angle laser light scattering and refractive index system.Dextran with similar molecular weight as polysaccharides was selected as the reference substance.Orthogonal experiment and single factor tests were used to optimize the pretreatment conditions for the determination of total polysaccharides in Polyporus.Polysaccharides in Polyporus with different areas and grades were determined by anthrone-sulfuric acid colorimetric method at 630 nm. Result: The linearity,stability,precision,repeatability and recovery rate of the established method all reached the standards,respectively.The content of total polysaccharides in samples from different areas ranged from 0.87% to 1.39%.The content of total polysaccharides in samples with different grades was 1.40% for first-grade pieces,1.21% for second-grade pieces, and 1.03% for third-grade pieces. Conclusion: The established method is simple,accurate and reproducible,and it can be used for the determination of polysaccharides in decoction pieces of Polyporus.The content of polysaccharides in samples from different origins varies greatly.The content of polysaccharides in samples with different grades shows a certain regularity.The content of polysaccharides is the highest in the first-grade pieces,followed by the content in the second-grade,and the lowest in the third-grade.The results can provide a reference for formulating limits for the content of total polysaccharides and the grade standard of decoction pieces of Polyporus.
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Objective To establish a method for content determination of total flavones and polysaccharides in Hedysari Radix in different producing areas in Gansu Province.Methods The contents of total flavones and polysaccharides in Hedysari Radix in different producing areas in Gansu Province were determined by ultraviolet spectroscopy with calycosin-7-glucoside and glucose as reference substance, and the wavelength was set at 260 nm and 484 nm.Results The contents were from 2.82 mg/g to 6.79 mg/g for total flavones and from 106.14 mg/g to 746.40 mg/g for total polysaccharides in Hedysari Radix in different producing areas in Gansu Province. The recoveries of total flavones and total polysaccharides were 97.96％ and 102.90％, respectively.Conclusion There was difference in contents of total flavones and polysaccharides of Hedysari Radix in different producing areas in Gansu Province, and the method of using ultraviolet spectroscopy is simple, reproducible, accurate and reliable, which can be preferably used as the method for content determination of total flavones and polysaccharides in Hedysari Radix.
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Objective: To optimize the extraction method for American ginseng. Methods: Based on the contents of the total poly-saccharides and saponins in American ginseng, the ethanol concentration, extraction duration, solid-liquid ratio and extraction time were selected to determine the factors and levels by single-factor test. And then 3 factors further considered were the concentration of ethanol, solid-liquid ratio and extraction time. Results: The optimum extraction process was adding 12-fold amount of 40% ethanol, and extracting for 3 times. Conclusion: The optimized extraction process is simple, quick and stable, and can be used to extract the total polysaccharide and saponins from American gingeng.
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Objective: To optimize the suitable technology of Ophiopogon japonicus (OJ) fumigation based on the changes of relative indexes after the different levels of sulfur fuming. Methods: The phenol-sulfuric acid method was used to determine the content of polysaccharide. HPLC-ELSD method was used to determine the contents of Ophiopogon saponins D (OPD) and D' (OPD') with the mobile phase of acetonitrile-water (50:50), drift tube temperature at 40℃, and nitrogen volumetric flow of 2.0 L/min. The fingerprints of OJ were determined by HPLC-UV method, the mobile phase was eluted with acetonitrile-0.1% phosphoric acid gradient, and the detection wavelength was 297 nm. The contents of ethylophiopogonanones A (MA) and B (MB) were determined using the same method. Using the contents of total polysaccharides of OJ, OPD and OPD', MA and MB, and the contents of other components in the fingerprints as control indexes, and the dosage of sulfur and the time of sulfur fumigation as factors, the central composite design-response surface methodology (CCD-RSM) was used to optimize sulfur fumigation process. Results: In the experimental design range, the content of polysaccharides increased first and then decreased with the increase of sulfur dosage and sulfur fumigation time, and the contents of OPD and OPD' increased slightly and then decreased. The contents of MA and MB were slightly reduced. The sulfur fuming process was determined as follows: sulfur dosage was 120-200 g/m3, sulfur fuming time was 0.6-1.5 h, binomial equation: Y = 0.574 + 0.054 3 X1-0.008 13 X2-0.518 X12 + 0.131X22 + 0.153 X1X2 (R-Sq = 0.971). Process validation: the absolute value of deviation was less than 3%. Conclusion: In this study, the changes of chemical composition of OJ were studied, and the sulfur smelting process suitable for OJ was established.
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Objective To establish the assay method for the total polysaccharide in Pudi Enema.Methods Phenol-sulfuric acid method was used for chromogenic reaction.The content of total polysaccharide was measured by UV spectrophotometry at 488.8 nm.Results The total polysaccharides calibration curve was at the range of 0~22.635 mg/L, with regression function being Y=0.062 06 X-0.003 34(r=0.999 8).The recovery of calycosin was 98.36%(RSD=2.34%).Conclusion This method is sensitive,rapid,accurate and reliable.It can be used to assay the content of total polysaccharide in Pudi Enema.
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AIM To study the interactions of component (group) compatibility of Sanwu Huangqin Decoction in incubation model of rat liver microsomes.METHODS With maackiain and ephedrine as internal standards,HPLC was adopted in the simultaneous content determination of total tlavonoids (baicalin,wogonoside,baicalein,wogonin and oroxylin A) from Scutellariae Radix and total alkaloids (oxymatrine,oxysophocarpine and matrine)from Sophorae flavescentis Radix in whole prescription component (group) compatibility and single medicinal material part (group) at seven time points (0,15,30,45,60,90 and 120 min),followed by the calculation of their in vitro metabolic rates.RESULTS In whole prescription component (group) compatibility,the contents of baicalin and wogonoside were first increased (0-0.5 h) and then decreased (0.5-2.0 h),those of baicalein,wogonin and oroxylin A showed increasing trends (more obvious within 0.5 h).The stable and gende metabolisms of various alkaloids reached balance within 20 min.CONCLUSION The bioavailability improvement and efficacy enhancement of total flavonoids from Scutellariae Radix may attribute to the compatibility with total alkaloids from Sophorae flavescentis Radix and total polysaccharides from Rehmanniae Radix in Sanwu Huangqin Decoction
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AIM To study the interactions of component (group) compatibility of Sanwu Huangqin Decoction in incubation model of rat liver microsomes.METHODS With maackiain and ephedrine as internal standards,HPLC was adopted in the simultaneous content determination of total tlavonoids (baicalin,wogonoside,baicalein,wogonin and oroxylin A) from Scutellariae Radix and total alkaloids (oxymatrine,oxysophocarpine and matrine)from Sophorae flavescentis Radix in whole prescription component (group) compatibility and single medicinal material part (group) at seven time points (0,15,30,45,60,90 and 120 min),followed by the calculation of their in vitro metabolic rates.RESULTS In whole prescription component (group) compatibility,the contents of baicalin and wogonoside were first increased (0-0.5 h) and then decreased (0.5-2.0 h),those of baicalein,wogonin and oroxylin A showed increasing trends (more obvious within 0.5 h).The stable and gende metabolisms of various alkaloids reached balance within 20 min.CONCLUSION The bioavailability improvement and efficacy enhancement of total flavonoids from Scutellariae Radix may attribute to the compatibility with total alkaloids from Sophorae flavescentis Radix and total polysaccharides from Rehmanniae Radix in Sanwu Huangqin Decoction
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Objective: To compare the content of total saponins and total polysaccharides between formula granule and traditional decoction of Sijunzi decoction.Methods: UV spectrophotometry was used to determine the content of total saponins and total polysaccharides in formula granule and traditional decoction of Sijunzi decoction respectively at the wavelength of 540 nm and 488 nm.Results: The absorbance of ginsenoside Re had a good linear correlation with the concentration within the range of 10.909-65.454 μg · ml-1 (r=0.999 7), and the average recovery was 98.49%(RSD=0.85%, n=6);the absorbance of D-anhydrous glucose had a good linear correlation with the concentration within the range of 2.160-12.960 μg · ml-1 (r=0.999 7), and the average recovery was 99.46%(RSD=0.73%, n=6).The contents of total saponins from 3 batches of formula granule were slightly higher than those from traditional decoction,and that of total polysaccharides in formula granule was slightly lower than that in traditional decoction,and there was no significant difference between the two groups (P>0.05)Conclusion: The difference of material basis between formula granule and traditional decoction of Sijunzi decoction is small, and formula granule is more feasible for clinical application.
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Objective: To establish rapid quantitative analysis models of multiple indicators for quality control in the concentrating process of Lvjiao Buxue Granule (LBG) by means of near infrared spectroscopy(NIRS). Methods: Thesolid content (SC), totalpolysaccharides concentration (TPC), and ferulic acid concentration (FAC) were determined by weighing method, phenol-sulfuric acid method, and HPLC method, respectively. Calibration models of them were established with partial least squares regression (PLSR)method. The established models were applied to predicting the unknown samples for testing the performance of the models. Results: The correlation coefficients (r) of SC, TPC, and FAC were all above 0.9450.The root mean square error in calibration (RMSEC) and root mean square error in prediction (RMSEP) were very close, and the relative standard errors of predictions (RSEP) were all less than 10%, indicating that the models predicted and performed well. Conclusion: NIRS technique combined with chemo metrics can provide a novel efficient and environmental approach for the fast simultaneous determination of key quality indicatorsfriendly, and havethe potential application value in the concentration process of LBG.
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Objective To study and establish the extraction technology of hypoglycemic components of total flavonoids and polysaccharides from Gynura divaricata (L .) DC ..Methods Based on the singer factor test ,the influences of extracting times ,diameter of drug powder ,soaking time ,solid-liquid ratio ,ethanol concentration ,and extracting time on extraction ratio of total flavonoids were studied .The best extraction technology of total flavonoids from Gynura divaricata (L .) DC .was es-tablished through L9 (3)4 orthogonal tests .On the base of previous studies combine with L9 (3)4 orthogonal tests ,the best ex-traction technology of total polysaccharides from Gynura divaricata (L .) DC .was established .Results The optimized extrac-tion technology of total flavonoids was confirmed as follows :drug powder of less than 10 mesh diameter were soaking 0 .5 hour with 80% ethanol ,and solid-liquid ratio 1∶20 ,and extracting twice ,each time one hour .The optimized extraction technology of total polysaccharides was confirmed as follows :add 20 times water to the residue of ethanol ,extraction temperature 90℃ , and extracting twice ,each time one hour .The extraction rate of both total flavonoids and polysaccharides were more than 80% .Conclusion The extraction technology of hypoglycemic components of total flavonoids and polysaccharides from Gynura divaricata (L .) DC .established in this study is stable ,reproducible ,and lays a foundation for the further studies on hypogly-cemic components in Gynura divaricata (L .) DC .
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Objective:To screen fine germplasm resources and suitable cultivated place for Paris polypyh lla var.yunnanensis by determining the content of total saponins , total flavonoids and total polysaccharides collected from Yunnan and Guizhou .Methods:An Ultraviolet-visible spectrophotometry method was applied to determine the total contents of saponin , flavonoid and polysaccharide in 14 germplasm resources of Paris polyhp ylla var.yunnanensis.Results:The contents of saponins, flavonoids and polysaccharides in Paris polyphylla var.yunnanensis collected from the different habitats were obviously different .The herb with high quality was selected from Xiaguan and Dali with the highest total content of saponins and the content of the three components was 55.760 0 and 61.632 0 mg· g-1 , respectively , which was significantly higher than that from the other habitats .Conclusion: It is proved that the quality of Pa ris polyphyll a var.yunnanensis produced in Dali of Yunnan is better than that from the other regions , and the high yield culture techniques should be studied further .
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OBJECTIVE:To investigate the correlation of total saponins and total polysaccharides content in the roots of Pari-dis rhizoma genus plants with Arbuscular mycorrhizal fungi(AMF)infection rate and rhizosphere soil nutrients. METHODS:Tak-ing 16 pieces of Paridis rhizoma herbs and rhizosphere soil in Three Gorges Reservoir Area as the research object,mycorrhizal in-fection rate and infection intensity,the contents of total saponins and total polysaccharides in roots and stem,and rhizosphere soil nutrient contents were all determined. SPSS18.0 software was adopted for data multiple comparison and correlation analysis. RE-SULTS:Paridis rhizoma genus plants roots in Three Gorges Reservoir Area could form a good symbiotic relationship with AMF. In-fection rate ranged from 85.19% to 99.80%,and infection intensity ranged from 16.55% to 72.27%. Paridis rhizoma from differ-ent origins and varieties had significant difference in the contents of total saponins and total polysaccharides (P<0.05). Soil pH was moderate,the average content of total nitrogen was in insufficient level,the average concentration of the remaining nutrients belonged to middle levels or above. Correlation analysis showed that the mycorrhizal infection rate and infection intensity of Paridis rhizoma were not significantly correlated with its quality. The total saponins were significantly negatively correlated with organic matter and available nitrogen. The total saponins and total polysaccharides were positively correlated with available phosphorus,po-tassium and pH value,but they were not significantly correlated with the other soil nutrients. CONCLUSIONS:Paridis rhizoma is selective to different soil nutrients. Rich soil available phosphorus and potassium supply are in favor of the synthesis and accumula-tion of total saponins and total polysaccharides in Paridis rhizoma roots.
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OBJECTIVE:To compare the differences in the contents of water-soluble total protein,total phenolic acid and total polysaccharides among the water decoctions of crude Cibotium barometz and processed products and to illuminate the effect of pro-cessing on 3 kinds of components of C. barometz. METHODS:UV-visible spectrophotometry was adopted to determine the con-tents of water-soluble total protein,total phenolic acid and total polysaccharides in the water decoction of crude C. barometz and 4 processed products,namely sand-scorch C. barometz,yellow wine C. barometz,salt C. barometz and steamed C. barometz,at wavelengths of 590,760 and 489 nm respectively. RESULTS:The contents of water-soluble total protein in 5 samples were 4.03%,3.32%,3.13%,3.33% and 3.49%,those of total phenolic acid therein were 0.25%,1.34%,1.38%,2.34% and 1.41%,and those of total polysaccharides therein were 28.56%,36.06%,45.21%,49.60% and 49.01%,respectively. CONCLU-SIONS:All above processing methods have an effect to some degree on the contents of the 3 kinds of components of C. barometz, where the contents of water-soluble total protein are lower after processing,while those of total phenolic acid and total polysaccha-rides are higher thereafter.
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This article was aimed to study enhancing efficacy and reducing toxicity on anti-tumor from total flavonoids and total polysaccharides in Taxus mairei compatibility with taxol. MTT assay was performed to analyze the cell proliferation and apoptosis of 4T1, A549, MCF-7 tumor cells after treatment of total flavonoids and total polysaccharides in T. mairei combined with taxol at different concentrations. Isobolograms was used to investigate the behavior of combination therapy. Therapy experiments were conducted on ICR mice bearing S180 sarcoma. The tumor weight was measured to calculate inhibition rate according to uniform design. Blood cells indicators were detected by blood cell analyzer. The results showed that total polysaccharides and total flavonoids can enhance the inhibition effect on 4T1, A549 cells by taxol. Total polysaccharides can enhance the inhibition effect of taxol to MCF-7 in vitro, and S180 sarcoma-bearing mice in vivo. It reduced the toxic effects of anti-tumor effect of taxol in vivo. It was concluded that taxol can achieve the effect of enhancing efficacy and reducing toxicity through combining with total flavonoids and total polysaccharides in T. mairei.
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OBJECTIVE:To optimize the technology of separation and purification of active constituents from Morus alba L. METHODS:With the adsorption rate and desorption rate as the indexes,static and dynamic macroporous resin adsorption-desorp-tion methods were adopted to optimize the type of macroporous resin,the loading amount of fluid,the volume fraction and amount of eluent in the separation and purification technology of total flavonids. With the yield and content of polysaccharide as the index-es,water extraction and alcohol precipitation method was used to optimize the volume fraction of alcohol for polysaccharide precipi-tation in the separation and purification technology of total polysaccharides. With the absorbance of total alkaloids as the index,ion exchange method was employed to optimize the loading amount of fluid and the amount of eluent in the separation and purification technology of total alkaloids. Then the verification was conducted. RESULTS:The optimal technology of total flavonids from Morus alba L. was with D101 macroporous resin as the adsorbent,loading fluid mass concentration of 0.1 g(material medicinal)/ml,loading fluid amount 9 times as the column volume(BV),70% alcohol of 5 BV for elution. The optimal volume fraction of alcohol for polysaccharide precipitation in the technology of total polysaccharides was 80%. The optimal technology of total alka-loids was with loading fluid amount of 3 BV,0.5 mol/L ammonia water of 8 BV for elution. After separation and purification,the purities of total flavonids,total polysaccharides and total alkaloids from Morus alba L. were 35.68%,47.14% and 55.79%,and the RSDs were 1.28%,1.61% and 1.14%(n=3). CONCLUSIONS:The optimal technology is stable and suitable for the separa-tion and purification of active constituents from M. alba L.
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Objective:To optimize the ethanol precipitation technique for Fufang Shenqi soft capsules. Methods: An orthogonal design was used to optimize the technique with the relative density of the concentrated solution, ethanol concentration, standing time, temperature of ethanol precipitation as the influencing factors and the yield of dry extract and the content of total polysaccharides as the indices. Results:The best ethanol precipitation technique was as follows:the relative density of the concentrated solution was 1. 10, 95% ethanol was used to obtain 60% ethanol concentration, and the standing time was 48 h under the temperature of 10-30℃. Con-clusion:The optimized ethanol precipitation technique for Fufang Shenqi soft capsules is simple and practicable, and suitable for prac-tical production.
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Paris fargesii is a famous Chinese traditional medicinal plant used as antipyretic, antidotal, antiphlogistic and analgesic. This research aimed at the determination of the total polysaccharides contents in P. fargesii, which were collected from Baoxing country of Sichuan Province in order to get the optimized extraction technology. Ultrasonic method was adopted in the extraction of total polysaccharides from P. fargesii. Through single factor experiment and orthogonal test, the effects of ultrasonic extraction conditions on the total polysaccharides extraction were measured, including solid-liquid ratio, ultrasonic temperature, ultrasonic power and ultrasonic time. Then colorimetric method was applied for the assaying. The optimized extraction conditions of total polysaccharides from P. fargesii were as follows: ultrasonic power: 600w, solid-liquid ratio: 1:20, ultrasonic extraction time: 1.5h and ultrasonic temperature: 90℃. Under the optimum parameters, the total polysaccharides extraction efficiency was 0.0715%.
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OBJECTIVE:To establish phenol-sulfuric acid method for the determination of the content of total polysaccharides in Weile granules.METHODS:The samples were deproteinized by Sevag method,colored by phenol-sulfuric acid method(water bath at 60 ℃ for 30 min),and the content of total polysaccharides in Weile granules was determined with Glucose as control.RESULTS:The linear range of Glucose was 20.47～102.36 ?g?mL-1(r=0.999 4),with an average recovery of 99.09%(RSD=0.92%,n=6).The content of total polysaccharides in Weile granules was 41.63%.CONCLUSION:This method was simple,accurate and reproducible,and suitable for quality control of Weile granules.