1. 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, and 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.
2. Components of the kit:
Each kit contains 2-in-1 Aflatoxin (B1,B2,G1,G2)/Ochratoxin A immunoaffinity columns and 1 instruction manual.
3. Necessary items not provided in the box:
- Equipment
- LC-MS(LC-MS/MS)
- Centrifuge capable of at least 3,000-4,000 x g
- Nitrogen gas evaporator apparatus
- Nitrogen gas tank and pressure regulator
- Air-pressure controller bracket
- Air pump
- Balance with 0.01g readability
- High-speed homogenizer (maximum speed> 10,000 r/ min) or shaker
- Grinder
- Sieving screen:2-mm
- Graduated cylinder: 100 mL/10 mL
- Funnel: 50 mL
- Syringe: 10 mL/20 mL
- Pipette: 1 mL and pipette tips
- Homogenization flask (or 250-mL conical flask with pestle)
- Vials and tubes
- Rapid qualitative filter paper
- Microfiber filter paper (e.g., Whatman 934-AH)
- Column holder and syringe connector plug (for use with immunoaffinity columns)
3. Reagents
- Methanol (CH3OH): Chromatographic Grade
- Methanol (CH3OH): Analytical Grade
- Acetic acid (CH3COOH): Chromatography Grade
- Sodium chloride (NaCl): Analytical Grade
- TWEEN-20 (C58H114O26): Analytical Grade
- Sodium hydroxide (NaOH): Analytical Grade
- Hydrochloric acid (HCl): Analytical Grade
- Distilled/deionized water
4. Note:
- Allow the immunoaffinity column to return to room temperature (22 to 25°C) before use.
- The affinity column should be stored at 2 to 8°C, do not freeze.
- Dot use any expired immunoaffinity column.
- The sample volume can be increased or decreased appropriately as required, and the volume of the extraction solution should be adjusted accordingly.
- Column capacity:
| Toxin name | Column Capacity: ng |
| OTA | 100 |
| AFT B1 | 300 |
| AFT B2 | |
| AFT G1 | |
| AFT G2 |
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 effects.
- WARNING: Aflatoxin and Ochratoxin A are toxic and carcinogenic; protective equipment such as gloves and masks should always be used during handling.
- 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 run.
