Method for measuring content of plant polysaccharide
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A plant polysaccharide and measurement method technology, applied in the detection field, can solve the problems of negative impact of the environment and experimenters, poor specificity, excessive addition of PMP, etc., and achieve the effect of shortening the hydrolysis time
Inactive Publication Date: 2021-10-01
LANZHOU INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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Problems solved by technology
After several years of application, implementation and inspection, the following shortcomings have been found in the production industry for the classic sulfuric acid-phenol method standard detection method: First, the accuracy of the detection results is poor, which is mainly due to the poor specificity and poor reproducibility of the method 1. Serious interference; Second, the detection method is cumbersome to operate, cannot achieve high-throughput detection, and is dangerous, because this method is a color reaction, and the detection result is directly related to the color development time, so it is difficult to realize automated and large-scale detection; The third is that this method uses a large amount of highly corrosive and toxic reagents such as concentrated sulfuric acid and phenol, which has a relatively large negative impact on the environment and experimenters.
[0004] The PMP derivatization method is to first hydrolyze the polysaccharide with sulfuric acid or trifluoroacetic acid, then derivatize it by PMP, and then use high-performance liquid chromatography to measure the content of the polysaccharide, but this method has the following disadvantages: one needs to be derivatized, and the operation is cumbersome; Excessive addition of PMP is not easy to remove. Generally, it is removed by multiple extraction methods. However, due to the large error caused by the operating habits of different experimenters, the results of the determination of plant polysaccharide content are inaccurate.
However, the traditional polysaccharide hydrolysis takes a long time, which is not conducive to the popularization of ion chromatography to determine the content of plant polysaccharides.
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Embodiment 1
[0037] 1. Extraction of Lycium barbarum polysaccharide
[0038] Take the dried medlar, remove impurities such as fruit stalks and leaves, crush, mix well and set aside.
[0039] Accurately weigh (0.5 ± 0.05) g of sample powder, add 100 mL of ether, heat to reflux for 1 hour, let stand, let cool, carefully discard the ether solution, and place the residue on a water bath to evaporate the ether. Add 100mL of 80% ethanol, heat to reflux for 1 hour, filter while hot, wash the filter residue and filter with 30mL of hot 80% ethanol, put the filter residue and filter paper in a flask, add 150mL of water, and heat to reflux for 2 hours. Filter while hot, wash the filter with a small amount of hot water, combine the filtrate and lotion, and concentrate the solvent by rotary evaporation, then transfer to a 25mL volumetric flask, wash the rotary evaporator with a small amount of hot water, combine the concentrated solution and lotion, let cool, Dilute to the mark with water and shake we...
Embodiment 2
[0080] Embodiment 2: Determination of Lycium barbarum polysaccharide content in Lycium barbarum by the established method
[0081] Take 20 μL of the sample solution after microwave-assisted digestion, inject it into the ion chromatograph, analyze according to the above ion chromatographic conditions, and record the peak area. Qualitative according to the retention time of the reference substance, and quantitative by the external standard method. The content of each monosaccharide in the measured 3 batches of wolfberry actual samples is shown in Table 7:
[0082] Table 7 The mass concentration table of each monosaccharide component in the actual sample of Lycium barbarum (unit: μg / mL)
[0083]
[0084] According to the test results, the content of Lycium barbarum polysaccharide in three batches of Lycium barbarum samples was 2.24%, 2.28% and 2.56%, respectively.
[0085] 7. Standard recovery experiment
[0086] At the same time, the recovery rate of standard addition was ...
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Abstract
The invention provides a method for measuring the content of plant polysaccharide, which comprises the following steps: degreasing a sample, removing small molecular compounds, performing reflux extraction with water, and performing microwave-assisted digestion to obtain plant polysaccharide hydrolysate. Monosaccharide in the hydrolysate is subjected to gradient elution through an ion exchange chromatographic column and a sodium hydroxide / sodium acetate solution and detected through a pulse ampere detector, the composition and content of monosaccharide generated by polysaccharide hydrolysis are measured, and the content of plant polysaccharide in the sample is calculated with the sum content of all monosaccharide as the standard. The method for measuring the content of the plant polysaccharide can effectively shorten the hydrolysis time of the plant polysaccharide, and then the content is measured by using the ion chromatography, so that a better choice is provided for the detection of the plant polysaccharide, and meanwhile, technical support and basic data are provided for controlling the quality of medicinal plants in plants and further researching the biological activity of the medicinal plants.
Description
technical field [0001] The invention belongs to the technical field of detection, and in particular relates to a method for measuring plant polysaccharide content. Background technique [0002] Plant polysaccharides are compounds composed of many identical or different monosaccharides with α- or β-glycosidic bonds. They are commonly found in plants in nature and are very important biological macromolecules in plants. They have various biological activities, such as regulating Immunity, regulating blood sugar and blood lipids, protecting the liver, anti-oxidation, regulating intestinal microorganisms, etc. Therefore, accurate determination of plant polysaccharide content and monosaccharide composition is of great significance for the study of its chemical structure and properties. [0003] At present, the determination methods of plant polysaccharide content mainly include sulfuric acid-phenol method, PMP derivatization method, ion chromatography and so on. The classic sulf...
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