Immunoaffinity Column for Aflatoxins (B1, B2, G1, G2) and Zearalenone IAC 2-in-1

1. Purpose:

The immunoaffinity column can selectively adsorb aflatoxin (B₁, B₂, G₁, G₂) and Zearalenone（ZEN） from the sample solution, thereby having a highly targeted purification effect on the sample. The sample solution purified by passing through the affinity column can be directly used for HPLC analysis or LC-MS/MS analysis after concentrating with nitrogen gas and resuspended.

2. Principle:

The basis of the measurement is the antigen-antibody reaction. Antibodies are connected to the column and the toxin in the sample is extracted, filtered, diluted, and then passed slowly through the 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. The toxins are then eluted with eluent and injected into an analytical instrument for detection.

3. Components of the kit:

Each kit contains 2-in-1 Aflatoxins (B1, B2, G1, G2)/Zearalenone immunoaffinity columns and 1 instruction manual.

4. Necessary items not provided in the box:

4.1 Equipment

• LC-MS(LC-MS/MS)
• Centrifuge capable of at least4000RPM
• 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:2-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)

4.2 Reagents

• Methanol (CH₃OH): ChromatographicGrade
• Acetonitrile (CH₃CN): AnalyticalGrade
• Acetic acid (CH₃COOH): ChromatographyGrade
• Sodium chloride (NaCl): AnalyticalGrade
• TWEEN-20 (C₅₈H₁₁₄O₂₆): AnalyticalGrade
• Sodium hydroxide (NaOH): Analytical Grade
• Hydrochloric acid (HCl): AnalyticalGrade
• Distilled/deionized water

 5. 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:

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
• 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 and Zearalenone are toxic and carcinogenic; protective equipment such as gloves and masks should always be used during
• Vessels and tools used to handle toxin solutions should be completely immersed in a sodium hypochlorite solution (5% v/v) overnight.
• Ensure the LC-MS/MS is clean, and the tubing is primed appropriately for each