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Coarse roots Rubus polysaccharide determination_98

By Stephen Hudson,2014-10-30 13:26
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Coarse roots Rubus polysaccharide determination_98

Coarse roots Rubus polysaccharide determination

     Author: Zhou Heng, Li Tianjiao, Hong Zhenfeng, Hu Juan

     Abstract Objective To establish the coarse roots Rubus method for the determination of polysaccharide. Methods phenol

    - sulfuric acid method color, the standard curve in the 490 nm light punishment Determination of crude polysaccharide content of the roots of Rubus. The results of the regression equation: Y = 0.8325X 0.022 3, r = 0.999 8, the measured coarse roots

    Rubus polysaccharide content 9.473%, the average recovery was 99.69%, RSD = 0.56% (n = 6). Conclusion The method is simple, precision and reproducibility are good, can be used for rough Rubus extraction and determination of polysaccharides.

     Key words rough Rubus; polysaccharide; phenol - sulfuric acid

    method; spectrophotometry

     Abstract: ObjectiveTo study the optimum extration process of polysaccharides from Rubus alceaefolius Poir. MethodsThe content of polysaccharids from Rubus alceaefolius Poir was

    determined by spectrophotometry at 490 nm. ResultsThe regression equation was Y = 0.832 5 × 0.022 3, r = 0.999 8.

    The percent of polysaccharide in Rubus alceaefolius Poir was 9.473%. The average recovery rate was 99.69% and RSD was 0.56% (n = 6). ConclusionThe method is proved to be simple, accurate and reliable, and it can be used to extract and determine the polysaccharides from Rubus alceaefolius Poir.

     Key words: Rubus alceaefolius Poir; Polysacharide; Determination

     Coarse leaves of dicotyledonous plants of Rubus Rosaceae,

    Rubus coarse roots, leaves can be used as medicine, with blood circulation Huayu, heat the efficacy of detoxification [1], can Kiyotoshi hot and humid, and so hot and humid due to intrinsic toxic silt removal. According to the Guangzhou

    military to prepare a "common herbs Manual" is written: "blood circulation Removes silt, heat to stop bleeding, cure acute and chronic hepatitis, hepatosplenomegaly, march hemoglobinuria, mastitis, trauma, bleeding, stomatitis." "Guangxi Chinese Herbal Medicine , "recorded:" heat, swelling, bleeding, loose silt. "Fujian Anxi in Fujian area are widely used in the treatment of various types of hepatitis, has a good basis for civil medication. In this paper, phenol -

    sulfuric acid Determination of coarse roots Rubus

    polysaccharide with satisfactory results.

     An instrument and Drug

     Coarse roots were collected from Rubus Fujian Anxi, Fujian College of Traditional Chinese Medicine through the Department of specialists was identified as dicotyledonous

    Rosaceae Rubus Rubus alceaefolius Poir; D anhydrous glucose

    (Lot No. :110833-200302, China Pharmaceutical and Biological Products The); 95% ethanol, concentrated sulfuric acid were of analytical grade, phenol (provisional preparation). UV-722

    spectrophotometer (Shanghai Analysis Instrument Factory); BP2110 Scale Electronic Analytical Balance (Sartorius Corporation); DHG 9240-type electric oven thermostat

    (Shanghai Experimental Equipment Co., Ltd. Jing Wang); RK52A rotary evaporator ( Shanghai Yali-Rong Biochemical Instrument

    Factory).

     2 Methods and Results

     2.1 Preparation of glucose standard solution Weigh Precision 105 ? dried to constant weight of glucose 50.52 mg, dissolved in distilled water and fixed volume to 100 ml shall

    be 0.505 2 mg / ml glucose standard solution.

     2.2 Preparation of phenol solution of phenol obtained 100 g, plus 0.1 g of aluminum and NaHCO3 0.05 g, distillation, collecting 182 ? fraction, saying that taking 7.5 g,

    dissolved in 150 ml water, brown bottle Fang Bingxiang back

    home.

     2.3 The precise amount of the standard curve drawing to take home 0.2,0.4,0.8,1.2,1.6,2.0 ml glucose standard solution of triangular flask with plug in, and each with distilled water to make into a 2.0ml, and then 5% phenol solution were

    added to 6.0 ml, shake , and then slowly added by drops of concentrated sulfuric acid 30.0 ml, to fully shake, heat, and then transferred to 50.0 ml flask, add distilled water to 50.0 ml constant volume at 30 ? water bath temperature 30 min,

    remove the cooled to room temperature. Another ditto with distilled water as a blank control operation, the absorbance was measured at 490 nm, rendering the standard curve to establish the regression equation: Y = 0.832 5X 0.022 3 r = 0.999 8, the linear range of 2.02 ~ 20.2 μg / ml.

     2.4 Rough Rubus Extraction and purification of polysaccharides [2] will be rough Rubus rooted in ESP 350-type

    grinder in the smash in powder, take rough Rubus powder 50 g placed in 1 000 ml flask, add water (solid liquor ratio 1:10)

    micro-boiling back 3 times, 1 h / times, leaching, merge extract, evaporation to 300 ml, by adding anhydrous ethanol makes the alcohol concentration of not less than 70%, precipitation of sedimentation, filtration may precipitate,

    was re-dissolved in water leaching liquid, with Sevag method to remove protein (repeat three times), plus alcohol concentration of ethanol to make up to 80%, and home refrigerator overnight put it aside to turn to the supernatant was precipitated, add ethanol After mixing leaching, may

    residues, residues with anhydrous ethanol, acetone and washed several times, dry deposition was pure polysaccharide. Reposted elsewhere in the paper for free download http://

     2.5 Determination of conversion factor

     Weigh accurately 65 ? drying to constant weight Rubus

    crude polysaccharide 20 mg, buy 100 ml flask, add water-

    soluble and is scheduled to scale capacity, shake, as the polysaccharide reserve liquid. Exact amount of liquid reserves polysaccharide obtained 0.10 ml, add water to 1.00 ml,

    measured according to the standard curve method of measuring the absorbance value, calculated according to the standard curve concentration. Conversion factor determined by the formula: f = W / CD formula: W is the weight polysaccharide

    (μg), C for the test solution is calculated according to the standard curve of a single sugar concentration (μg * ml-1), D

    for the polysaccharide dilution factor, measured at f = 2.595 (n = 6).

     2.6 stability test taking sample solution, standard

    solution and purification of polysaccharide solution of 2.0 ml, according to the standard curve prepared under way since the "plus-phenol reagent 6.0 ml", the absorption measured in accordance with law degrees, and, respectively, 0,5 , 10,15,20,25,30,35,40,50,60 min measurement, analysis stability. The results of sample solution absorbance, standard

    solution absorption, absorption of refined polysaccharide solution RSD (%) respectively, 0.98,1.02,0.86, we can see, the sample liquid at least 1 h within the measurement is stable and reliable.

     Experimental precision precision 2.7 Rubus said crude

    polysaccharides obtained 25 mg, according to the sample solution preparation and content of the sample under measurement method is to measure the absorbance, measured six consecutive times, average 0.634, RSD 0.37%, indicating precision Good.

     2.8 to reproduce the experiment to take rough Rubus polysaccharide six copies, according to the sample solution preparation method, respectively, were prepared by six mass concentration of 0.1 mg / ml solution, in accordance with the sample solution preparation and content of the sample under

    measurement method is determined by absorption degree of value, average 0.645, RSD 1.66%, reproducibility was good.

     2.9 Determination of content of the sample

     Precision Weigh rough Rubus powder 25 mg, with 80%

    ethanol soaked overnight, and then in the Soxhlet extractor using 80% ethanol, reflux 2 h, leaching, abandoned to the filtrate, residue continue to distilled water, reflux extraction 2 h, leaching, the filtrate washed repeatedly with the filter paper to the 250 ml flask, the constant volume, shake derived sample solution. When measured, precision drawing sample solution 0.20 ml, add water to 1.00 ml, according to the standard curve of measured absorbance values were measured in the same way, determined by the formula

    polysaccharide content (in crude polysaccharide dollars): polysaccharide content = CDf × 100 / W. Where: C samples, the

concentration of glucose solution (μg * ml-1), D for the

    sample solution dilution factor, f is conversion factor, w is the weight of sample (μg), the measured results of 9.473%.

    The results in Table 1. Table 1 Samples of polysaccharide (omitted)

     2.10 recovery experiment

     Precision drawing sample solution 0.2 ml (6 copies) each with precision by adding the same amount of standard solution,

    depending on the sample determine the absorbance were measured to calculate the average recovery rate of 99.69%, RSD 0.56%. The results in Table 2. Table 2 recovery experiment (omitted)

     3 Discussion

     In this study, with water extraction of crude extraction and alcohol precipitation Rubus polysaccharide, using Sevage law, excluding protein purified polysaccharide with phenol -

    sulfuric acid colorimetric determination of sugar content, equipment required for this method is generic, drug prices are

    low, Operation is also easy, through the stability, precision, reproducibility and recoveries experimental results can be seen that this method is suitable for rough determination of Rubus polysaccharide. UV spectrophotometer, through access to

    a large number of literature choose maximum wavelength of 490 nm was measured, from the experimental results of view, the purified crude Rubus 9.473% polysaccharide content of the testing results for the first time reported for the drug

    Quality Standards provide a scientific basis.

     References

     [1] YAO Zhen-sheng, Yang Liang Wu. Jiangxi Province Rubus medicinal plants and the use of the proposed [J]. Chinese herbal medicine, 1995,18 (11): 551.

     [2] Li Yikui. TCM pharmacology test methods [M].

    Shanghai: Shanghai Science and Technology Press, 1999:461. Reposted elsewhere in the paper for free download http://

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