A kind of tobacco leaf multi-in-one heavy metal rapid quantitative detection card and detection method

A quantitative detection and heavy metal technology, applied in the field of heavy metal detection, can solve the problems of inaccurate detection results, low detection efficiency, and low detection sensitivity of the detection system, and achieve the effects of improving reaction sensitivity, reducing antagonism, and reducing antibody dosage.

Active Publication Date: 2022-08-02
成都安普诺生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] However, since colloidal gold can only be effectively combined with antibodies in a weakly alkaline environment, once the pH of the detection object is not weakly alkaline, it cannot be accurately detected, and the detection of heavy metals needs to be extracted with strong acid, although by increasing the dilution factor It can be barely detected, but the detection results are not accurate
[0010] At present, the heavy metal colloidal gold products on the market are mainly based on a single qualitative item, which has the advantages of rapidity, good specificity, and high sensitivity. However, traditional colloidal gold detection cards are generally used for single item (single heavy metal) detection, using two-line method mode , that is, the quality control line C and the detection line T. The gold-labeled antibody is a single-item antibody, so the test card can only detect one item, and cannot detect multiple items and quantitative analysis. It is difficult to adapt to the current detection of heavy metals. The detection of a single heavy metal species is developing towards the simultaneous detection of multiple heavy metal components; and the existing heavy metal quantitative detection system does not take masking measures, which cannot effectively interfere with the chelation process of natural chelating agents in complex samples of agricultural products such as tobacco. Masking, while the detection system has low detection sensitivity, cannot detect multiple heavy metals at the same time, and the detection efficiency is low

Method used

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  • A kind of tobacco leaf multi-in-one heavy metal rapid quantitative detection card and detection method
  • A kind of tobacco leaf multi-in-one heavy metal rapid quantitative detection card and detection method
  • A kind of tobacco leaf multi-in-one heavy metal rapid quantitative detection card and detection method

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Experimental program
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Embodiment 1

[0059] The embodiment of the present invention provides a preparation method of modified colloidal gold, comprising the following preparation steps:

[0060] 1.1 Take 100 mL of 0.01% chloroauric acid aqueous solution and heat it to boiling, accurately add 1.8 ml of 1% trisodium citrate aqueous solution under stirring, the golden yellow chloroauric acid aqueous solution turns red within 2min, continue to boil for 15min, that is, 40nm colloidal gold solution;

[0061] 1.2 Take 100mL of colloidal gold solution with a diameter of 40nm, add 400-800μL of 0.1M K 2 CO 3 Adjust the pH value of the colloidal gold solution at 7.5, add HS-PEG20000-COOH solution with a concentration of 1% at a concentration of 0.01-1% of the mass of the colloidal gold solution, stir at 200 r / min for 1 h, and centrifuge at 8000 r / min for 5 minutes to remove the supernatant. The supernatant and the lower centrifuge are modified colloidal gold.

[0062] In the preparation method provided in this example, t...

Embodiment 2

[0066] The embodiment of the present invention provides a preparation method of a gold-labeled antibody, comprising the following preparation steps:

[0067] 2.1 Activation

[0068] 1) Take 100 mL of the modified colloidal gold solution obtained in Example 1 into a triangular flask, slowly add 40 mL of MES buffer solution at a constant speed, mix with a vortex mixer for 5 min, and then centrifuge at 10000 r / min for 5 min to remove the supernatant. Observe the reconstitution situation; add 100mL MES, resuspend by ultrasonic for 1-3min, ultrasonic power 50%, observe the reconstitution situation;

[0069] 2) Add 5 mg of EDC [1-(3-dimethylaminopropyl)-3-ethylcarbodiimide] and 5 mg of NHS (N-hydroxysuccinimide) to the centrifuge tube respectively, and then add 5 mg of NHS (N-hydroxysuccinimide). Add 1mL MES buffer solution and mix well to obtain EDC solution and NHS solution, which are prepared and used now, preferably away from light;

[0070] 3) Take 0.2 mL of the NHS solution ob...

Embodiment 3

[0078] The embodiment of the present invention provides an all-in-one heavy metal rapid quantitative detection card, and the structure of the detection card refers to figure 1 and figure 2 , the detection card includes a detection test paper placed in the card case, the detection test paper includes a bottom plate 1, the bottom plate 1 is a plastic bottom plate, and the bottom plate 1 is sequentially pasted from one end to the other end with a sample pad 2, containing a variety of heavy metals to be detected corresponding to One end of the sample pad 2 is crimped on one end of the binding pad 3, and the other end of the One end of the chromatographic membrane and one end of the absorbent paper 6 close to the chromatographic membrane 4 are crimped on one end of the chromatographic membrane 4, and the absorbent paper 6 is a absorbent filter paper; the chromatographic membrane 4 is coated with There are multiple detection lines T (T1, T2, T3, etc.) and quality control lines C. ...

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Abstract

The invention belongs to the field of heavy metal detection, and relates to a tobacco leaf multi-in-one heavy metal rapid quantitative detection card and a detection method. The gold-labeled antibody formed by the combination of modified colloidal gold and an antibody is stable and reduces the time when the gold-labeled antibody reacts with the antigen. The antagonism between the antibodies can effectively improve the reaction sensitivity and reduce the amount of the antibody, and effectively overcome the immune reaction during detection; the multi-in-one heavy metal rapid quantitative detection card using the gold-labeled antibody of the present invention can simultaneously detect a variety of heavy metals, and realize the realization of a variety of heavy metals. The rapid detection and quantification greatly simplifies the operation process and improves the detection efficiency.

Description

technical field [0001] The invention belongs to the field of heavy metal detection, and relates to a tobacco leaf multi-in-one heavy metal rapid quantitative detection card and a detection method. Background technique [0002] The original meaning of heavy metals refers to metals with a specific gravity greater than 5. In terms of environmental pollution, heavy metals mainly refer to heavy elements with significant biological toxicity such as mercury (Hg), cadmium (Cd), lead (Pb), chromium (Cr) and metalloid arsenic (As). Heavy metals cannot be biodegraded, on the contrary, they can be enriched thousands of times under the bio-magnification of the food chain, and finally enter the human body. Heavy metals can interact strongly with proteins and enzymes in the human body, making them inactive and accumulating in some organs of the human body, causing chronic poisoning. The main toxic effects are anemia, neurological disorders, kidney damage, severe softening of bones, diges...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/58G01N33/577G01N33/558G01N33/543G01N33/53G01N33/52
CPCG01N33/587G01N33/577G01N33/558G01N33/54346G01N33/5306G01N33/5308G01N33/52
Inventor 吴晓迪米军罗超刘畅刘松洁黄磊潘虹
Owner 成都安普诺生物科技有限公司
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