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Method for determining activity of alpha-amylase inhibitor by colorimetric method via resin de-coloring

A technology of amylase inhibitor and colorimetry, which is applied in the direction of color/spectral characteristic measurement, etc., can solve the problems of vacancy and inaccurate determination of active ingredients, and achieve the effect of reducing loss, reducing color interference, and good effect

Inactive Publication Date: 2014-03-12
GUANGXI MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When the color of the sample liquid is light, the background color subtraction method is directly used to eliminate interference; when the color of the sample liquid is dark, activated carbon is generally used to decolorize the Chinese herbal medicine extract to eliminate interference. Deeper extracts need more activated carbon to decolorize. Activated carbon decolorization will cause serious adsorption of active ingredients and result in inaccurate determination results. How to take an effective method to remove pigments in Chinese herbal medicine extracts and reduce color contrast color analysis interference, to ensure the accuracy of the measurement results to the greatest extent, this is a problem that needs to be solved
Chinese patent application 201110414922.3 uses highly polar polystyrene copolymer macroporous resin ADS-7 combined with neutral alumina to decolorize the total saponins of Panax notoginseng, and patent application number 200810153792.0 uses anion exchange resin as the decolorization medium to decolorize the water-soluble extract of Salvia miltiorrhiza For decolorization, these are only decolorization of the extract, and there is no report on the use of colorimetric analysis
Patent Application No. 200410014122.2 The use of D101 macroporous resin is only used to improve the extraction rate of effective parts of Astragalus membranaceus, and the research on the use of D101 macroporous resin for colorimetric analysis of Chinese medicine extracts after decolorization has been vacant so far

Method used

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  • Method for determining activity of alpha-amylase inhibitor by colorimetric method via resin de-coloring
  • Method for determining activity of alpha-amylase inhibitor by colorimetric method via resin de-coloring

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Resin decolorization is used for the method of colorimetric assay α-amylase inhibitor activity:

[0035] Weigh 10.00g of black tea powder, add 100mL distilled water, extract in 90°C water bath for 30min, filter, add 80mL distilled water to medicinal residues, extract in 90°C water bath for 30min, filter, add 60mL distilled water to extract in 90°C waterbath for 30min, filter, three times The filtrates were combined, and the volume was adjusted to 250mL (concentration is equivalent to 0.04g crude drug per 1ml). Take 30mL of the filtrate and centrifuge (4000r / min) for 10min, and take 20mL of the supernatant as the sample solution.

[0036]Take D101 macroporous resin after conventional treatment. Take a certain amount of D101 macroporous resin and use filter paper to absorb the surface moisture, then weigh 8.00g of the macroporous resin to remove the surface moisture, and put it into a 2cm*15cm chromatography column with a column volume of about 13mL. Take the centrifuge...

Embodiment 2

[0042] According to the operation method of Example 1, Morinda officinalis, Radix Astragali, and Rhodiola were decolorized with D101 macroporous resin, and the fractions after decolorization were collected. Comparison of the color depth of the elution fractions of different gradients, determination of A 520 The results are shown in Table 4.

[0043] Table 4. Comparison of deliquified colors of Morinda officinalis, Astragalus, and Rhodiola and their inhibitory effects on a-amylase

[0044]

[0045] The results showed that the medicinal material became lighter in color after being eluted by D101 macroporous resin, and the inhibitory effect on α-amylase was concentrated in the 70% and 95% fractions with lighter color.

Embodiment 3

[0047] Resin decolorization is used for the method of colorimetric assay α-amylase inhibitor activity:

[0048] Accurately weigh 10.00g of Prunella vulgaris powder, add 100mL of deionized water, extract in a 93°C water bath for 30min, filter, add 80mL of deionized water to the residue, extract in a 93°C water bath for 30min, filter, add 60mL of deionized water to the residue in a 93°C water bath Extract for 30 minutes, filter, combine the three filtrates, and dilute to 250mL (concentration: 0.04g / mL). Take 30mL of the filtrate and centrifuge (4000r / min) for 15min, and take 20mL of the supernatant as the sample solution. Accurately weigh 8.00 g of D101 macroporous resin (the same batch of pre-treated D101 macroporous resin as in Example 1) after removing the moisture on the surface of the resin, and put it into a chromatographic column with a specification of 2cm*15cm. The column volume is about 13mL. Take the centrifuged supernatant and load the sample, control the flow rate...

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Abstract

The invention discloses a method for determining the activity of an alpha-amylase inhibitor by a colorimetric method via resin de-coloring. According to the method, the interferences on the colorimetric analysis and detection of the activity of alpha-amylase by pigments of a traditional Chinese medicine extracting solution can be effectively reduced; a method which has the advantages of simplicity, practicability, environmental protection and good analysis repeatability is provided for screening the alpha-amylase inhibitor in vitro. The method comprises the following steps: crushing plants or medicinal materials; extracting by water or alcohol; combining filtering solutions and centrifuging a certain quantity of filtering solution; after centrifuging, taking a liquid supernatant; enabling the liquid supernatant taking as sampling liquid to pass through a D101 macroporous resin column; carrying out gradient eluting by using ethanol-water solutions with different concentrations; and respectively collecting flow parts with different gradients to obtain the flow parts which have different color degrees and have different activities on the alpha-amylase inhibitor. According to the method, the D101 macroporous resin column is used for de-coloring a medicinal material extracting solution so as to relieve the interferences on the colorimetric analysis by a color of the inhibitor in the process of determining the activity of the alpha-amylase inhibitor.

Description

technical field [0001] The invention relates to a method for measuring the activity of alpha-amylase inhibitors, in particular to a method for measuring the activity of alpha-amylase inhibitors by colorimetric method for resin decolorization. After the extract of Chinese herbal medicine was decolorized by D101 macroporous resin, the inhibitory effect of the decolorized fraction on a-amylase activity was determined by colorimetry, which was used as a reference for the method of screening a-amylase inhibitors of Chinese herbal medicine in vitro. Background technique [0002] α-amylase inhibitors can effectively inhibit the activity of saliva and pancreatic α-amylase, hinder the decomposition and digestion of carbohydrates in food, and reduce the production and intake of glucose. Clinically, α-amylase inhibitors can effectively prevent and treat diabetes and hyperlipidemia, and can also be used in the development of health care drugs for weight loss. Therefore, screening and s...

Claims

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Application Information

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IPC IPC(8): G01N21/25
Inventor 龙盛京许云青姚丹媚
Owner GUANGXI MEDICAL UNIVERSITY
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