Test strip for detecting aristolochic acid A and aristolochic acid B in traditional Chinese medicine and preparation method of test strip

An aristolochic acid and test strip technology, which is applied in biological tests, measuring devices, material inspection products, etc., can solve the problems of being unsuitable for rapid screening by quality inspection departments and suitable for laboratory testing, and is easy to popularize on a large scale. Low cost of application and detection, good stability

Pending Publication Date: 2021-06-01
NANCHANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Aiming at the deficiencies of the prior art, the present invention provides a test strip for detecting aristolochic acid A and B in traditional Chinese medicine and a preparation method thereof, which solves the problem that the existing detection method of aristolochic acid A and B is only suitable for Laboratory testing requires professionals to operate, and is not suitable for on-site rapid screening of quality inspection departments and consumers

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  • Test strip for detecting aristolochic acid A and aristolochic acid B in traditional Chinese medicine and preparation method of test strip

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Experimental program
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Effect test

Embodiment 1

[0036] Preparation of test strips for detecting aristolochic acids A and B in traditional Chinese medicine:

[0037] (1) Preparation of AAs monoclonal antibody-colloidal gold marker

[0038] ①Preparation of colloidal gold: dilute 1% chloroauric acid to 0.01% with ultrapure water, put in an oil bath with magnetic stirring, heat to boil, add 10mL 1% trisodium citrate for every 500mL 0.01% chloroauric acid, and continue to boil , Stop heating until the liquid turns red, and replenish the evaporated water after cooling to room temperature to obtain a colloidal gold solution.

[0039] ② Preparation of AAs monoclonal antibody-colloidal gold marker

[0040] Under magnetic stirring, add K to the colloidal gold solution 2 CO 3 The pH of the solution was adjusted to 6.0, 1.75 μg of anti-AAs monoclonal antibody was added to the colloidal gold solution, and the stirring reaction was continued for 15 minutes. Add 10% BSA to a final concentration of 1.0%, and continue stirring for 15 mi...

Embodiment 2

[0048] Detection of total AAs in traditional Chinese medicine samples:

[0049] (1) Sample detection: Weigh the crushed Fangji in a conical flask, add 10-20 times of methanol as the extract, shake well for 3-10 minutes, filter with qualitative filter paper, and collect the filtrate. Take 1mL of the filtrate, dilute it 4-7 times with 0.05-0.1mol / L PBS solution containing 0.5%-5% Tween-20 fully dissolved after stirring, mix well and use it as the sample solution, take 100μL and drop it in the example 1 Test on the sample pad of the prepared test strip, and observe the test result after 10 minutes.

[0050] (2) Result analysis: when both the detection line and the quality control line are red, it means that the sample is negative, that is, the content of aristolochic acid A and B in the sample is less than 6.0ng / mL; when only the detection line is red, it means that the sample is positive, That is, the content of aristolochic acid A and B in the sample is greater than 6.0ng / mL; ...

Embodiment 3

[0052] Detection of various indicators of test strips:

[0053] (1) Validity testing of test strips

[0054] Aristolochic acids A and B were weighed into conical flasks, dissolved in methanol to prepare a 6.0ng / ml solution, and an appropriate amount of PBST buffer was added to fully dissolve 0.05-0.1mol / L containing 0.5%-5 % Tween-20 PBS solution, take 100 μL for detection, when the detection line and quality control line are red, it means that the sample is negative, that is, the content of aristolochic acid A and B in the sample is less than 6.0ng / mL; The red line indicates that the sample is positive, that is, the content of aristolochic acid A and B in the sample is greater than 6.0ng / mL; when only the detection line develops color or neither the quality control line nor the quality control line develops color, it means that the test strip is invalid.

[0055] (2) Sensitivity experiment

[0056] Adopt the method described in step (1), detect the AAs standard substance of...

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Abstract

The invention relates to the technical field of medicine detection, in particular to a test strip for detecting aristolochic acid A and aristolochic acid B in traditional Chinese medicine and a preparation method of the test strip. The test strip comprises a nitrocellulose membrane, a sample pad, a conjugate release pad, a water absorption pad and a bottom plate, wherein the nitrocellulose membrane comprises a detection antigen coated with aristolochic acid and bovine serum albumin coupling and a goat anti-mouse IgG antibody, the conjugate release pad is coated with an aristolochic acid monoclonal antibody-colloidal gold marker, and the detection sensitivity of the test strip reaches 6.0 ng / mL. The test strip has the characteristics of high specificity, high sensitivity, simplicity and convenience in operation, rapidness and easiness in detection of aristolochic acid in a traditional Chinese medicine sample, and can be widely popularized and applied.

Description

technical field [0001] The invention relates to the technical field of drug detection, in particular to a test strip for detecting aristolochic acids A and B in traditional Chinese medicines and a preparation method thereof. Background technique [0002] Aristolochic acids (English: Aristolochic acids, referred to as "AAs"), also known as aristolochic acids, phagocytic acids or aketolin, are a class of nitrophenanthrene carboxylic acids. Aristolochic acids A and B are mainly found in Aristolochiaceae plants such as Aristolochia and Asarum. Among the aristolochic acid compounds, the main toxic components are aristolochic acid Ⅰ and aristolochic acid Ⅱ (ie, aristolochic acid A and B), which are partially reduced to aristolochic acid under the catalysis of nitroreductase. The other part of papilolactam further interacts with DNA during the reduction process to form adducts, which are mutagenic and carcinogenic. At present, besides Guan Mutong, which has been withdrawn from th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/58G01N33/577G01N33/558G01N33/53G01N33/532G01N33/531
CPCG01N33/587G01N33/577G01N33/558G01N33/5308G01N33/532G01N33/531
Inventor 黄志兵韩全斌谢建华黄婷李利锋
Owner NANCHANG UNIV
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