- Overview:
Aflatoxins are toxic metabolites of a class of fungi (such as Aspergillus flavus and Aspergillus parasiticus). They are highly carcinogenic and are found mainly in grains, peanuts, nuts, cottonseeds, animal feed, vegetable oils, as well as animal tissues and blood. Among them, Aflatoxin B1 (AFT B1) ranks first in terms of toxicity, carcinogenicity and frequency of contamination. Aflatoxin M1 is a hydroxylated metabolite of Aflatoxin B1 and a strong carcinogen. Cow milk and its products are prone to Aflatoxin M1 contamination.
Specification: 3mL,25T;
Samples: Grain and oil, nuts, feed, herbs, tea, condiments, dairy products, etc.
- 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. Aflatoxins are then eluted with methanol and injected into an analytical instrument for detection.
- Storage conditions and period of validity
Storage Conditions: 2 to 8°C
- Components of the kit:
Each kit contains Aflatoxin immunoaffinity columns of various specifications and 1 instruction manual.
- Necessary items not provided in the box:
Equipment
HPLC/LC-MS/MS
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 RPM) or shaker
High-speed freezing centrifuge (can be set to 10°C, 7, 500 RPM)
Centrifuge tube:50-mL
Constant temperature incubator or water bath (can be set to 35–37℃)
Grinder
Sieving screen:1-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)
- Reagents
Methanol (CH3OH): Analytical grade for extraction/chromatography grade for elution
Acetonitrile (CH3CN): Analytical Grade
Sodium chloride (NaCl): Analytical Grade
TWEEN-20 (C58H114O26): Analytical Grade
Hydrochloric acid (HCl): Analytical Grade
Sodium hydroxide (NaOH): Analytical Grade
Potassium chloride (KCl): Analytical Grade
Potassium dihydrogen phosphate (KH2PO4): Analytical Grade
Disodium hydrogen phosphate dodecahydrate (Na2HPO4·12H2O): Analytical Grade
Distilled or deionized water
- 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 |
AFT B1 AFT B2 AFT G1 AFT G2 | 300 |
AFT M1 | 100 |
AFT M2 | 100 |
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 is 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.
Follow appropriate instrument precautions if using HPLC.