Immunoaffinity Column for Deoxynivalenols (DON/15-ADON)

1. Purpose:

The immunoaffinity column can selectively adsorb Deoxynivalenol (Vomitoxin, DON) and 15-acetyldeoxynivalenol (15-ADON), and has a highly targeted purification effect. Samples that have passed through the column for purification may be used in HPLC, LC-MS/MS and other devices for analysis after concentrating with nitrogen gas.

Affinity columns can be used in combination with HPLC or LC-MS/MS to achieve rapid testing, and to increase signal-to-noise ratio and improve the accuracy of the detection method.

2. Overview:

DON belongs to the single-endomycin family of molds and is produced by certain Fusarium fungi. 15-ADON is derivative of DON. These toxins are mostly distributed in grain seeds such as wheat, barley, and corn. Due to their high cytotoxicity and immunosuppressive properties, they pose a health threat to humans and animals.

3. Principle:

The basis of the measurement is the antigen-antibody reaction. Antibodies are connected to the column and the aflatoxin in the sample is extracted, filtered, and diluted, and then passed slowly through the aflatoxin immunoaffinity column. The toxins bind to the antibodies in the column and the immunoaffinity column is then washed to remove other unrelated substances that have not been bound. Deoxynivalenol is then eluted with methanol and injected into an analytical instrument for detection.

4. Components of the kit:

Each kit contains Deoxynivalenol immunoaffinity columns of various specifications and 1 instruction manual.

5. Necessary items not provided in the box:

Equipment

HPLC/LC-MS/MS
Nitrogen gas evaporatorapparatus
Nitrogen gas tank and pressureregulator
Air-pressure controller bracket
Air pump
Balance with 0.01greadability
High-speed homogenizer (maximum speed> 10,000 r/ min) or shaker
Grinder
Sievingscreen:1-mm
Graduated cylinder: 100 mL/10mL
Funnel: 50mL
Syringe: 10 mL/20mL
Pipette: 1 mL and pipettetips
Homogenization flask (or 250-mL conical flask withpestle)
Vials andtubes
Rapid qualitative filterpaper
Microfiber filter paper (e.g., Whatman934-AH)
Column holder and syringe connector plug (for use with immunoaffinitycolumns)

5.2 Reagents

Methanol (CH₃OH): Analytical grade forextraction/chromatography grade for elution
Acetonitrile (CH₃CN): AnalyticalGrade
Sodium chloride (NaCl): AnalyticalGrade
Potassium dihydrogen phosphate (KH₂PO₄): AnalyticalGrade
Disodium hydrogen phosphate dodecahydrate(Na₂HPO₄12H₂O): Analytical Grade
Polyethylene glycol (8000Da, HO(CH₂CH₂O)_nH): AnalyticalGrade, to improve filtration speed, if the supernatant is obtained by centrifugation , this reagent can be
TWEEN-20 (C₅₈H₁₁₄O₂₆): AnalyticalGrade
Hydrochloric acid (HCl): AnalyticalGrade
Sodium hydroxide (NaOH): AnalyticalGrade
Potassium chloride (KCl): AnalyticalGrade
Distilled or deionizedwater

6. Note:

Allow the immunoaffinity column to return to room temperature (22 to 25°C) before
The affinity column should be stored at 2 to 8°C, do not
Dot use any expired immunoaffinity
The sample volume can be increased or decreased appropriately as required, and the volume of the extraction solution should be adjusted
Columncapacity

Toxin name	Column Capacity:(ng)
DON	2000
15-ADON	2000

When the content of the toxin in the sample divided by the dilution factor is higher than the column capacity, it is necessary to reduce the volume of the sample solution appropriately, and retest.

The pH of the loading solution of the affinity column should be within the range of 6 to 8. If it deviates from this range, the pH should be adjusted with dilute hydrochloric acid or dilute sodium hydroxide.
Maintaining consistency (such as polarity, pH, and concentration) between the test solvent loaded into any analytical instrument and the mobile phase can help eliminate any adverse solvent
WARNING: Aflatoxin is toxic and carcinogenic; protective equipment such as gloves and masks should always be used during handling.
Vesselsand tools used to handle toxin solutions should be

completely immersed in a sodium hypochlorite solution (5% v/v) overnight.