1. Overview:
Ochratoxin A (OTA) is a toxic metabolite produced by some species of Aspergillus and Penicillium,it is a mycotoxin with high kidney and liver toxicities, and has teratogenic, mutagenic, and carcinogenic effects.
2. Principle:
The basis of the measurement is the antigen-antibody reaction. Antibodies are connected to the column and the Ochratoxin A in the sample is extracted, filtered, and diluted, and then passed slowly through the Ochratoxin A 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. Ochratoxin A is then eluted with methanol and injected into an analytical instrument for detection.
3. Components of the kit:
Each kit contains Ochratoxin A immunoaffinity columns of various specifications and 1 instruction manual.
4. Necessary items not provided in the box:
- Equipment
- HPLC
- Centrifuge capable of at least 3,000-4,000RPM
- Nitrogen gas evaporatorapparatus
- Nitrogen gas tank and pressureregulator
- Air-pressure controllerbracket
- Airpump
- 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. Whatman 934-AH)
- Columnholder and syringe connector plug (for use with immunoaffinity columns)5.1 Reagents
- Methanol (CH3OH): Analytical grade forextraction/Chromatography grade for elution
- Disodium hydrogen phosphate dodecahydrate (Na2HPO4∙ 12H2O):Analytical Grade
- Acetic acid (CH3COOH): ChromatographyGrade
- Potassium dihydrogen phosphate (KH2PO4): AnalyticalGrade
- Sodium chloride (NaCl): AnalyticalGrade
- TWEEN-20 (C58H114O26): AnalyticalGrade
- Hydrochloric acid (HCl): Analytical Grade
- Sodium hydroxide (NaOH): AnalyticalGrade
- Potassium chloride (KCl): AnalyticalGrade
- Sodium Bicarbonate (NaHCO3):Analytical Grade
- Distilled or deionizedwater
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
- Column capacity: 100 ng, 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
- 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: Ochratoxin A 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
- Follow appropriate instrument precautions if using
