Time-resolved fluorescence immunochromatographic test strip for quantitative detection of capsaicinoids and preparing method and application thereof

A technology of time-resolved fluorescence and immunochromatography test paper, which is applied to the analysis of materials, measuring devices, instruments, etc., can solve the problems of high degree of purification, expensive instruments, cumbersome processing, etc., and achieve the effect of simple operation and high accuracy

Active Publication Date: 2016-06-08
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The existing capsaicinoids analysis and detection methods are mainly precision instrument analysis methods, mainly including high performance liquid chromatography and chromatography-mass spectrometry. These methods have high sensitivity and good accuracy, but they are expensive and require capsaicinoids. The degree of sampl

Method used

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  • Time-resolved fluorescence immunochromatographic test strip for quantitative detection of capsaicinoids and preparing method and application thereof
  • Time-resolved fluorescence immunochromatographic test strip for quantitative detection of capsaicinoids and preparing method and application thereof
  • Time-resolved fluorescence immunochromatographic test strip for quantitative detection of capsaicinoids and preparing method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1 Obtaining of anti-capsaicinoid monoclonal antibody

[0036] The anti-capsaicinoid monoclonal antibody is a monoclonal antibody secreted by the hybridoma cell line YQQD8 with the preservation number CCTCCNO.C201534. The specific preparation method:

[0037] The hybridoma cell line YQQD8 was injected into BALB / c mice treated with Freund's incomplete adjuvant in advance, the ascites of the mice was collected, and the antibody was purified by octanoic acid-ammonium sulfate method. Rat ascites, 4°C, centrifuged at 12000r / min for 30min, sucked the supernatant, mixed the obtained ascites supernatant with 3 times the volume of acetate buffer, slowly added n-octanoic acid under stirring, the volume of n-octanoic acid required per milliliter of ascites was 33μL, mix at room temperature for 30min, stand at 4°C for more than 2h; centrifuge at 12000r / min for 30min at 4°C, discard the precipitate, filter the obtained supernatant with double-layer filter paper, add 1 / 10 o...

Embodiment 2

[0064] Example 2: Time-resolved fluorescent immunochromatographic test strip for quantitative detection of capsaicinoids and its preparation

[0065] A time-resolved fluorescent immunochromatographic test strip for the quantitative detection of capsaicinoids, which includes a fluorescent test strip, an anti-capsaicinoids monoclonal antibody containing europium, a reaction bottle, a sample diluent, and a pipette for the sample diluent. The fluorescent test strip described above includes cardboard, and one side of the cardboard is pasted with water-absorbing pads, detection pads and sample pads sequentially from top to bottom. Adjacent pads are overlapped and connected at the joints, and the overlapping length is 1mm, wherein: the water-absorbing pad is 38mm long , 3.8mm wide; the detection pad is 25mm long and 3.8mm wide; the sample pad is 15mm long and 3.8mm wide. The detection pad is based on a nitrocellulose membrane, and a horizontal quality control line and a detection lin...

Embodiment 3

[0095] Example 3: Time-resolved fluorescent immunochromatographic test strip for quantitative detection of capsaicinoids and its application

[0096]A time-resolved fluorescent immunochromatographic test strip for quantitative detection of capsaicinoids, which includes a fluorescent test strip, a sample reaction bottle containing europium-labeled anti-capsaicinoids monoclonal antibody, a sample diluent and a pipette for the sample diluent, The fluorescent test strip includes cardboard, and one side of the cardboard is pasted with water-absorbing pads, detection pads and sample pads in sequence, and adjacent pads are overlapped and connected at the joints, and the overlapping length is 1mm, wherein: the length of the water-absorbing pads 41mm, width 4mm; detection pad length 25mm, width 4mm; sample pad length 13mm, width 4mm. The detection pad is based on a nitrocellulose membrane, and a horizontal quality control line and a detection line are arranged on the nitrocellulose mem...

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Abstract

The invention relates to a time-resolved fluorescence immunochromatographic test strip for quantitative detection of capsaicinoids and a preparing method and application thereof. The time-resolved fluorescence immunochromatographic test strip comprises a fluorescence test strip, europium-labeled anti-capsaicine, dihydrocapsaicin, olepresin capsaicin universal monoclonal antibody reagent and a reaction flask, wherein the fluorescence test strip comprises a paperboard, a water absorption mat, a detection mat and a sample mat adhere to the viscous surface of the paperboard from top to bottom in sequence, the adjacent mats are connected in an overlapping mode, a nitrocellulose membrane serves as the base mat of the detection mat, a horizontal quality control line and a detection line are arranged on the nitrocellulose membrane from top to bottom, the quality control line is coated with a rabbit antimouse polyclonal antibody, the detection line is coated with a capsaicinoids envelope antigen, and anti-capsaicine, dihydrocapsaicin, olepresin capsaicin universal monoclonal antibody are generated through secretion of a hybridoma cell strain with the preservation number of CCTCC NO.C201534. The test strip can be used for quantitative detection of the content of capsaicinoids, operation is easy and quick, and accuracy is high.

Description

technical field [0001] The invention relates to a time-resolved fluorescent immunochromatographic test strip for quantitatively detecting capsaicinoids, a preparation method and an application thereof. Background technique [0002] Capsaicin compounds are the main chemical substances that make peppers pungent, and their components mainly include capsaicin, dihydrocapsaicin, homodihydrocapsaicin, nordihydrocapsaicin, homocapsaicin and other compounds. Wherein capsaicin and dihydrocapsaicin account for about 90% of the total amount of capsaicinoids, and provide about 90% of the hot stimulation. Synthetic capsaicin has a similar structure and function to natural capsaicin, and is more advantageous than natural capsaicin in terms of price and spiciness. It is also widely used in medicine, insecticides, antifouling paints, military weapons and other fields. In addition to giving food a spicy taste and being used as a condiment, capsaicinoids also have anti-inflammatory and analg...

Claims

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Application Information

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IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 李培武杨青青马飞张奇张良晓张文李慧丁晓霞张兆威
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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