Immunoaffinity column for detecting vomitoxin and application of immunoaffinity column

A technology of vomitoxin and immunoprophylaxis, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of poor experimental sensitivity and repeatability, serious injury to operators, cumbersome operation process, etc., and achieve short pretreatment time, simple storage, The effect of high sensitivity

Active Publication Date: 2015-04-29
BEIJING KWINBON BIOTECH
View PDF6 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Traditional mycotoxin purification methods mostly use organic solvent extraction and chemical adsorption. These methods have many shortcomings: low specificity, low purification efficiency, and poor purification effect; poor experimental sensitivity and repeatability; Long time and high labor intensity; the use of organic solvents is likely to cause environmental pollution and cause greater harm to operators

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Immunoaffinity column for detecting vomitoxin and application of immunoaffinity column
  • Immunoaffinity column for detecting vomitoxin and application of immunoaffinity column
  • Immunoaffinity column for detecting vomitoxin and application of immunoaffinity column

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Preparation of immunoaffinity column for detection of vomitoxin

[0036] The preparation method of this immunoaffinity column mainly comprises the following steps:

[0037] 1) Prepare the solid phase medium coupled with vomitoxin antibody, i.e. CNBr-Sepharose-4B-Ab

[0038] 2) Load the solid phase medium into the column, and assemble the immunoaffinity column according to the combination and connection method of the affinity column.

[0039] The following step-by-step detailed description:

[0040] 1. Preparation of vomitoxin hapten

[0041] Mix 0.10g vomitoxin in 2mL dimethyl sulfoxide (DMSO), slowly add dropwise 0.1mL 1,3-propanediamine and 0.1mL pyridine in 2mL DMSO mixture at 60°C, continue the reaction after the dropwise addition is complete After 12h, the solvent and unreacted propylenediamine were removed by rotary evaporation, and the propylenediamine monocondensate of vomitoxin was quantitatively obtained. The synthetic route is as follows: figur...

Embodiment 2

[0059] Example 2 Detection of vomitoxin residues in samples

[0060] 1. Sample pretreatment

[0061] Homogenize the sample with a homogenizer; weigh 2.0±0.05g of the sample into a 50ml polystyrene centrifuge tube, add 16ml of deionized water, vortex with a vortexer for 5min, or shake on a shaker for 20min, above 3000g, room temperature (20 -25℃-77 / 68℉) centrifuge for 5min; filter all supernatant with qualitative filter paper for later use;

[0062] 2. Operation steps of sample passing through the column

[0063] Connect the immunoaffinity column under a 10ml syringe, and connect the affinity column product with an adapter; accurately draw 8ml (equivalent to 1g sample) of the test solution into the syringe, and use the syringe to control the flow rate of the test solution for 2 seconds Each drop passes through the affinity column; rinse the affinity column once with 5ml of deionized water, use a syringe to control the flow rate to 0.5-1 second per drop, and drain the liquid i...

Embodiment 3

[0064] Embodiment 3 sample detection example

[0065] 1. Blank determination of negative samples: the general matrix category includes (cereals, finished feed).

[0066] Take the blank samples of the above-mentioned various matrices (corn, soybean meal, wheat bran, distiller's grains, batch materials, concentrated materials), purify them through the vomitoxin immunoaffinity column, and perform instrumental detection to obtain the following results (see Table 1), each Samples were measured in triplicate.

[0067] Table 1 Various blank sample detection results

[0068]

[0069] The results show that the blank value detected by various matrices is far less than the detection limit of 1 μg / kg required by the state, indicating that the affinity column can be applied to the detection of samples of various matrices, and different sample matrices will not cause blank interference to the detection.

[0070] 2. Determination of accuracy and precision: Add vomitoxin at a concentrati...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses an immunoaffinity column for detecting vomitoxin and application of the immunoaffinity column. The immunoaffinity column comprises a solid-phase extraction column hollow column (1), an upper sieve plate (2), an upper cover (3), a lower cover (4), a solid-phase medium (5) and a lower sieve plate (6), wherein the solid-phase medium is coupled with a vomitoxin antibody. The invention further provides a method for detecting vomitoxin residues in grains and feed by adopting the immunoaffinity column for detecting vomitoxin. The immunoaffinity column disclosed by the invention has the characteristics of simplicity in operation, high sensitivity, high detection speed, low cost and the like, and is suitable for screening a lot of samples and monitoring in the field.

Description

technical field [0001] The invention relates to an immunoaffinity column for detecting vomitoxin and its application, in particular to an immunoaffinity column for detecting vomitoxin, which is especially suitable for sample pretreatment in the detection of vomitoxin residues in grains and finished feed. Background technique [0002] Vomitoxin (vomitoxin), also known as deoxynivalenol (deoxynivalenol, DON), is one of the trichothecene olefins, which is usually produced by growing in cereals (such as wheat, corn, barley and hay grass) ) produced by mycofusarin. Toxic effects of vomitoxin include: vomiting, reluctance to eat, gastroenteritis, diarrhea, immunosuppression, and hematological disorders. [0003] DON exists widely in the world, mainly polluting cereal crops such as wheat, barley, and corn, and also contaminating food products. Humans and animals can produce a wide range of toxic effects after eating grains and cereals contaminated by this toxin. In recent years, ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/544G01N30/60
CPCG01N30/60G01N33/544
Inventor 冯才伟罗晓琴万宇平何方洋常平平冯才茂杨昌松徐念琴
Owner BEIJING KWINBON BIOTECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap