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3,5 - dinitrosalicylic acid colorimetry Astragalus polysaccharide

2ml, 5 copies each, were placed in 10ml graduated test tube with stopper, according to The results mean total sugar absorption 0.524, RSD% = 1.06% (n = 5); average absorption of monosaccharides 0.231, RSD% = 1.37% (n = 5). Shows good precision of this Law. 2.3.5 repeatability take the same batch of sample preparation from crude 5 copies of the test solution,[link widoczny dla zalogowanych], to determine the content. Results polysaccharide samples averaged 51.1%, RSD% = 1.92%. 2.3.6 Test samples of sample recovered from crude accurately weighed (batch: 070408) 5 copies, each of about 17.5mg,[link widoczny dla zalogowanych], adding reference substance precise amount, determined according to determination results showed that the recovery of this Law well, see Table 2. Table 2,[link widoczny dla zalogowanych], recoveries of polysaccharide sample test results 2.3.7 Determination precise amount of total sugar solution and a single sugar solution 2ml, were placed in 10ml graduated test tube with stopper, according to 4 batches of samples for determination of polysaccharide, the results in Table 3. Table 3 from crude polysaccharide containing ■ 3 3.1 Determination discuss the choice of method of determination of polysaccharides, as well as a phenol sulfuric acid method, anthrone sulfuric acid method, the polysaccharide content of the two methods, you can not get rid of monosaccharide composition of the interference of high levels of determination than the actual results, and both methods use sulfuric acid, sulfuric acid intense heat water, which is difficult to control the color conditions, affect the determination results, leading to instability in the linear study, high concentrations of has serious corrosive sulfuric acid, inconvenience operation. Compared with the previous two methods, 3,5 dinitrosalicylic acid method or two more stable reagents, simple, rapid, high sensitivity, in addition to the interference of a single sugar component, the results are accurate, can be widely used. 3.2 3.2.1 Colour reaction time investigated the conditions of precision drawing reference solution 0.8ml, 5 copies in parallel, placed in 10ml graduated stoppered test tube, add water to make into a 2ml, precision or two nitrate added 3,5 salicylic acid test solution 2.0ml, mixing, heated water bath at 1000C 1,[link widoczny dla zalogowanych],3,5,7,9 min, remove immediately with ice-water bath to cool to room temperature, add water, 3.0ml. Shake well, with the corresponding reagent as blank absorbance were determined at 500nm. 1min reaction absorbance is very small, the reaction 3min later, the absorbance did not change the basic, comprehensive consideration, select the reaction time of 5min. 3.2.2 Precision temperature control solution absorb 0.8ml, 5 copies in parallel, placed in lOml graduated stoppered test tube, add water to make into a 2ml,[link widoczny dla zalogowanych], precision or two nitro acid test solution add 3,5 2. Oml, mixing, respectively, 60,70,80,9 O ℃ and heated in boiling water bath 5min, remove immediately with ice-water bath to cool to room temperature, add water 3. Oml, shake, made with the corresponding reagent blank absorbance were determined at 500nm. 60,70,80 ~ C reaction temperature, the absorbance of the lower, 90 ~ C and the boiling water bath, but the difference is much higher absorbance, considering, choose the boiling water bath for the reaction temperature.
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