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Antibody, method and reagent kit for detecting and determining chlorogenic acid

A chlorogenic acid and antibody technology, applied in chemical instruments and methods, specific peptides, measuring devices, etc., can solve the problems of lack of chlorogenic acid antibodies, time-consuming and labor-intensive, hindering research, etc.

Inactive Publication Date: 2009-07-22
BEIJING UNIV OF CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, biological samples containing chlorogenic acid (including plasma, urine, saliva, etc.) contain a large number of proteins and endogenous substances that affect the determination of the content; at the same time, chlorogenic acid may be in a combined state and must be processed to eliminate endogenous impurities and The interference of metabolites makes the combined chlorogenic acid free before it can be measured; at the same time, it needs to be concentrated to meet the requirements of instrument detection sensitivity, the processing procedure is complicated, time-consuming and laborious; at the same time, after the chlorogenic acid enters the body, the distribution in the tissue is Extremely small amount, even after enrichment, it is still extremely difficult to determine the content
In addition, for the distribution of chlorogenic acid in target organs and tissues, as well as the study of cellular and subcellular localization, methods such as immunohistochemistry and western-blot are required, but the lack of antibodies to chlorogenic acid hinders this research

Method used

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  • Antibody, method and reagent kit for detecting and determining chlorogenic acid
  • Antibody, method and reagent kit for detecting and determining chlorogenic acid
  • Antibody, method and reagent kit for detecting and determining chlorogenic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] hapten compound 1

[0035] Take 100g of honeysuckle medicinal material, use 95% ethanol or 50% ethanol aqueous solution, the solid-to-liquid ratio is 1:10, adjust the pH of the solution to 3, and extract 3 times at about 50°C. Combine the extracts, concentrate until there is no alcohol smell, refrigerate overnight at 4°C, filter with suction to obtain the supernatant, add the supernatant to polyamide resin and put it in a glass column (400×10mm) until the effluent becomes cloudy, and then Gradient elution with water, 10%, 20%, 30%, 40% and 50% ethanol aqueous solution (adjust pH to about 4.0 with hydrochloric acid), HPLC detection combined eluate with chlorogenic acid content greater than 60%, concentrated and placed at 0 ℃ refrigerator crystallization, suction filtration. The above crystals were dissolved in an appropriate amount of water and added to the top of the Sephadex LH-20 column. The chromatographic conditions were 50% acetone aqueous solution (pH=4) as the ...

Embodiment 2

[0037] According to sulfoxide chloride (SOCl 2 ) and chlorogenic acid into a three-necked flask, heated to reflux for 1 h under magnetic stirring, and distilled off excess chlorogenic acid to obtain the product chlorinated chlorogenic acid. According to the feeding ratio of γ-aminobutyric acid: CA=1.5:1, put γ-aminobutyric acid into the three-necked flask, add an appropriate amount of NaOH solution under ice bath, stir and dissolve, dissolve in dioxane, and add it to the constant pressure dropping funnel. Add this solution dropwise five times, and then add an appropriate amount of NaOH solution to keep the pH value of the reaction solution at 8-10. After the addition is complete, continue to react at 0-4°C for 5h. Rise to room temperature, adjust the pH value of the reaction solution to about 4.0 with concentrated hydrochloric acid, extract with ethyl acetate, combine the ethyl acetate extracts, wash with water, dry over anhydrous sodium sulfate, concentrate to obtain a solid,...

Embodiment 3

[0039] According to the method of Example 2, hexaalkylated chlorogenic acid was synthesized with 6-aminocaproic acid.

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Abstract

The invention provides a hapten comprising chlorogenic acid or derivatives thereof, an immunogen of the hapten which can be combined with the carrier material endowed with antigenicity, a conjugate (peridium source) of the hapten which is combined with tag reagent, and an antibody which can resist the immunogen; and the antibody can be combined with the determining position of at least one structural antigen in complete chlorogenic acid. The invention also provides a method and a kit used for testing or measuring the content of the chlorogenic acid in samples, and the usage of the conjugate and the antibody in the tested or measured chlorogenic acid. The invention has specificity to the chlorogenic acid, and can be used for testing the presence of the chlorogenic acid and measuring the content of the chlorogenic acid.

Description

technical field [0001] The invention relates to an antibody of a small molecular hapten and its preparation method and application, in particular to an antibody of chlorogenic acid and its derivatives and its preparation method and application. Background technique [0002] The present invention relates to methods and kits for detection and quantification of chlorogenic acid, and haptens, immunogens, conjugates and antibodies used therein. [0003] Where "detection" is the presence or absence of a specified analytical substance. [0004] Where "determination" refers to the quantitative analysis of a substance. [0005] Chlorogenic acid (Chlorogenic acid, hereinafter referred to as CA), molecular formula: C16H18O9, molecular weight: 354.30, is a depsipolic acid generated from caffeic acid (Caffeic acid) and quinic acid (Quinic acid, 1-hydroxyhexahydrogallic acid). Synonymous coffee tannic acid, systematic name 1,3,4,5-tetrahydroxycyclohexanecarboxylic acid-(3,4-dihydroxycin...

Claims

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Application Information

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IPC IPC(8): C07K14/76C07K14/765C07K14/435C07K16/44C07H17/07G01N33/577
Inventor 王庆国赵琰屈会化任会明蒋兴凯杨爱玲李翼飞
Owner BEIJING UNIV OF CHINESE MEDICINE
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