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Qualitative and quantitative analysis method for polyoses

A technique for quantitative analysis and polysaccharides, applied in the field of qualitative and quantitative analysis of polysaccharides, can solve the problems of high equipment requirements, poor practicability, and difficulty, and achieve the effects of improving sensitivity, separation effect and ultraviolet absorption characteristics

Active Publication Date: 2009-09-23
李绍平
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method will overcome the deficiencies of the prior art, such as cumbersome and time-consuming, high equipment requirements, great difficulty, and poor practicability.

Method used

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  • Qualitative and quantitative analysis method for polyoses
  • Qualitative and quantitative analysis method for polyoses
  • Qualitative and quantitative analysis method for polyoses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031]Extraction of polysaccharides from traditional Chinese medicine: Weigh 0.5g each of 9 kinds of Chinese herbal medicine powders of ginseng, Panax notoginseng, American ginseng, Cordyceps sinensis, Cordyceps militaris, Chizhi, Zizhi, Astragalus and Angelica, add 20 times the weight-to-volume ratio of water, and extract at 100°C high temperature water for 1 hours, the extract was centrifuged to obtain a supernatant, and 5 mL of the supernatant was added with absolute ethanol to a final concentration of 75%, and left to settle overnight at 4°C. After centrifugation, the precipitate was washed twice with 4 mL of 95% ethanol at 60°C. The alcohol was evaporated to dryness on a water bath, the residue was redissolved with 5 mL of hot water (60°C), and the supernatant was centrifuged to obtain the polysaccharide extract. Using the phenol-sulfuric acid colorimetric method as a quantitative reference, adjust the extract to a relative sugar content of 0.15 mg / mL (calculated as glucos...

Embodiment 2

[0039] Extraction of polysaccharides from traditional Chinese medicines: Weigh 0.5 g of the medicinal powders of Panax notoginseng and Astragalus membranaceus, and the rest of the operations are the same as the extraction steps of polysaccharides in Example 1.

[0040] Enzymatic hydrolysis of polysaccharide: same as embodiment 1.

[0041] HPSEC-ELSD analysis: same as Example 1.

[0042] PMP derivatization of polysaccharide hydrolyzate: after centrifugation of polysaccharide hydrolyzate of traditional Chinese medicine, take 600 μL of supernatant, mix it with an equal volume of ammonia water, then add 200 μL of 0.5mol / LPMP methanol solution, mix well and seal, and react at 70°C for 30 minutes. Take it out and let it cool to room temperature, add 2mL of water, shake well, evaporate to dryness at 50°C, and repeat the operation twice. The residue was dissolved in 1 mL of water, extracted with an equal volume of chloroform, and repeated three times, and the water layer was passed t...

Embodiment 3

[0046] Extraction of polysaccharides from traditional Chinese medicine: Weigh 0.5 g each of American ginseng, Cordyceps militaris and Cordyceps militaris medicinal material powders, and the rest of the operations are the same as the polysaccharide extraction steps in Example 1.

[0047] Enzymatic hydrolysis of polysaccharide: same as embodiment 1.

[0048] HPSEC-ELSD analysis: same as Example 1.

[0049] PMP derivatization of polysaccharide hydrolyzate: same as Example 2.

[0050] HPLC-DAD-MS analysis: same as embodiment 2.

[0051] Test results, refer to Table 2, Figure 5 to Figure 10 : American ginseng, Cordyceps sinensis and Cordyceps militaris have similar hydrolysis response characteristics to 13 glycosidases. The enzymatic hydrolysis products of the three, taking the pectinase enzymatic hydrolysis as an example, after derivatization with PMP, HPLC-DAD and HPLC-MS analysis can obtain characteristic spectra for identification.

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Abstract

The invention relates to a qualitative and quantitative analysis method for polyoses, which comprises the following steps: (1) the zymohydrolysis and the verification of the polyoses: measuring the content of a polyoses water solution by a colorimetric method; (2) comparing the change of a polyoses mapping before and after zymohydrolysis by HPSEC-ELSD analysis; (3) saccharide ingredient direct HPLC analysis of a polyoses zymohydrolysis solution or HPLC analysis after pre-column derivatization: identifying polyoses mapping and chromatogram peaks by taking a standard monosaccharose, a uronic acid, a disaccharide, and the like as contrasts and taking a chromatogram peak mass-to-charge ratio as reference, and using stable characteristic sections as a quantitative analysis index; and (4) realizing the qualitative and quantitative analysis of the polyoses by establishing polyoses zymohydrolysis responding characteristics and polyoses zymohydrolysis mapping which are based on structural information through the independent or combining application of the steps. The qualitative and quantitative analysis method can be used for the identification and quantitative measurement of the polyoses of traditional Chinese medicine, such as panax, panax pseudoginseng, gen-seng, aweto, cordyceps, ganoderma lucidum, fomes japonica, milk veteh, angelica, and the like and provides an effective method for controlling the quality of the polyoses and the products thereof.

Description

technical field [0001] The invention relates to a qualitative and quantitative analysis method for polysaccharides. Background technique [0002] Polysaccharides are a class of polycarbohydrates formed by condensing and dehydrating more than 10 monosaccharides in the form of glycosidic bonds. As an essential component of all living organisms, it is closely related to energy metabolism and various physiological functions to maintain life. In recent years, polysaccharides derived from plants, marine organisms and fungi have been widely used due to their remarkable biological activities such as anti-tumor [Lin Menggan, et al. Chinese Herbal Medicine, 2007, 38: 949-953.] .Shi Zhen Guo Yi Guo Yao, 2008, 19: 99-101.], hypoglycemic [Luo Zuyou, et al. Food Science, 2007, 28: 596-600.], antioxidant [Chen Xuan, et al. Chinese Journal of New Drugs, 2007, 16:1000-1004.] and anti-inflammatory and anti-viral [Ding Baojin, et al. Chinese Journal of Pharmaceutical Sciences, 2004, 39:561-5...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N30/89G01N30/02G01N30/72G01N21/78C12Q1/34
Inventor 李绍平管佳
Owner 李绍平
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