Simple and effective mulberry leaf polysaccharide detection method
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A mulberry leaf polysaccharide and detection method technology, applied in the detection field, can solve the problems of poor stability, cumbersome steps, poor reproducibility, etc., and achieve the effects of easy operation, high polysaccharide content, and simple extraction process
Inactive Publication Date: 2016-07-27
SOUTHWEST UNIV
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Problems solved by technology
However, the separation and purification of mulberry leaf polysaccharides with higher purity is cumbersome and has poor reproducibility; while the results of the determination of total sugar by the anthrone-sulfuric acid method will also be affected by proteins with higher tryptophan content, and the stability is poor; The experimental conditions of the method are strict (for example, the requirements for phenol are very high), and the determination process is limited by various conditions, so the reproducibility is poor
Therefore, none of the existing detection methods can reflect the content of polysaccharides in mulberry leaves simply, accurately and efficiently.
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Embodiment 1
[0042] A method for detecting polysaccharides from mulberry leaves, comprising the following steps:
[0043] (1) Extraction of mulberry leaf reducing sugar and water-soluble sugar
[0044] a. Weigh 1.00g of mulberry leaf powder into a 250mL conical flask, add 60mL of distilled water (first use a small amount of water to make the mulberry leaf powder into a paste), stir, bathe in 50°C water for 20min, and record it as extract A;
[0045] b. Transfer the extract A (including residue) obtained in step a to a 100mL volumetric flask in turn, rinse the conical flask with distilled water for 2-3 times, transfer the rinse solution to the volumetric flask, and then add 3mL of zinc acetate solution with a concentration of 219g / L and 3mL of potassium ferrocyanide solution with a concentration of 106g / L, and dilute to 100mL with distilled water, and record it as extract B;
[0046] c. Use a single-layer nylon cloth to filter the extract B obtained in step b, and collect the filtrate into...
Embodiment 2
[0084] The preparation of related reagents, the extraction method of mulberry leaf reducing sugar, water-soluble sugar and total sugar are the same as in Example 1.
[0085] Glucose standard solution preparation: The glucose standard solution was prepared according to the method in Example 1, with a final concentration of 1.056 mg / mL.
[0086] Glucose standard curve preparation: take 11 10mL colorimetric tubes, add 0.00mL, 0.08mL, 0.16mL, 0.24mL, 0.32mL, 0.40mL, 0.48mL, 0.56mL, 0.64mL, 0.72mL, 0.80mL glucose solution ( Add 0.00mL glucose solution as blank control), add water to dilute to 0.80mL, then add 0.60mL DNS reagent, shake well, boil in water bath for 5min, immediately cool to room temperature with cold water after taking out, then add distilled water to each tube to the mark (10mL), Shake well, and measure the absorbance value of each tube at a wavelength of 540nm. Taking the glucose content (mg) as the abscissa and the absorbance value as the ordinate, draw a standar...
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Abstract
The invention discloses a simple and effective mulberry leaf polysaccharide detection method. The method specifically comprises steps as follows: mulberry leaf water-soluble carbohydrates and mulberry leaf total carbohydrates are extracted, hydrolyzed and converted into reducing sugar, the light absorption values of a mulberry leaf water-soluble carbohydrate extracting solution and a mulberry leaf total carbohydrate extracting solution in the position of 540 nm are measured with a 3,5-DNS acid (dinitrosalicylic acid) colorimetric method, the mass of the mulberry leaf water-soluble carbohydrate extracting solution and the mulberry leaf total carbohydrate extracting solution in a measurement liquid are calculated on the basis of glucose according to the glucose standard curve, the content of mulberry leaf water-soluble carbohydrates and the content of mulberry leaf total carbohydrates (on the basis of reducing sugar) are calculated, and the content of mulberry leaf polysaccharides is calculated according to the formula that the quantity of the mulberry leaf polysaccharides (%) is equal to the product obtained by multiplying the difference between the quantity of mulberry leaf total carbohydrates (%) and mulberry leaf water-soluble carbohydrates (%) by 0.9. By means of the method, the problem that the color developing condition is difficult to control due to the fact that sulfuric acid releases much heat when meeting water is solved, besides, the measurement process is not affected by monosaccharide components, the operation is simple and safe, the result is stable and reliable, and the method is suitable for detecting the content of the polysaccharides in massive mulberry leaf samples.
Description
technical field [0001] The invention belongs to the detection field, and in particular relates to a simple and effective detection method for mulberry leaf polysaccharide. Background technique [0002] Mulberry leaves are the leaves of Morus L. (Morusalba L.) plants in the Moraceae family. They contain protein, inorganic salts, vitamins, polysaccharides, polyphenols, flavonoids, alkaloids and other functional ingredients. The Ministry of Health List it as a medicinal and edible Chinese herbal medicine. [0003] Modern pharmacological studies have shown that mulberry leaf polysaccharide, as one of the active components of mulberry leaves, has a significant hypoglycemic effect. At present, the detection methods of polysaccharide content in mulberry leaves are mainly anthrone-sulfuric acid method and phenol-sulfuric acid method. The anthrone-sulfuric acid method can determine almost all carbohydrates, and the phenol-sulfuric acid method is mainly used for the determination of...
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