Scigiene T-2 Toxin Immunoaffinity Column Instructions

Scigiene T-2 Toxin Immunoaffinity Column

Application

This product is used for quantitative detection of T2 by using Scigiene Lateral Flow Test Kits or other Mycotoxin tests.
It is suitable for purification of T2 in samples such as cereal, feed and food, in order to reduce matrix interference and thus improve analysis accuracy. The purified extraction solution can be detected by Scigiene Lateral Flow Test Kits or other Mycotoxin tests.

Product Performance Column

1. 1. Capacity：≥1500 ng/vial Recovery

2. Rate：85%~110%
3. Column Gel: Agarose Gel
4. Advantages Large loading capacity, Monoclonal antibody site-specific conjugation, Easy to elute,
5. High recovery rate.

Measuring Principle

Determination of the T-2 toxin (T2) immunoaffinity column is based on the antigen-antibody reaction. T2 monoclonal antibody is coupled to agarose gel material. After extracting, filtering, and diluting T2 in the sample, the sample extraction solution slowly passes through the Immunoaffinity Column. T2 in sample extraction solution combines with a specific monoclonal antibody, meanwhile, impurities are eluted from immunoaffinity column along with washing solution. Finally, T2 is eluted by using methanol.

Product Composition

• * 20 vial/box

  • 1* manual

Sample Preparation and Purification

1. 1. Weigh 20g pulverized sample, then add 100mL 80% methanol-water (V/V), and then vortex mix them in homogenizer for 2 minutes or vibrate for 30 minutes and extract later.

2. Centrifuge at 5000r/min for 5 minutes or filter through glass fiber filter paper. Later, take 10mL filtrate, and then add 20mL water to dilute and mix well.

3. Filter by rapid qualitative filter paper, then collect the filtrate.

4. Take 15mL filtrate for passing through the immunoaffinity column.
   Note:

5. For sample extraction solution (pH<6 or pH>8), it is necessary to adjust the pH value to neutral (pH 7) and use glass microfiber filter paper/quantitative filter paper.

6. For samples that are difficult to filter because of turbidity, centrifuge samples for separation.

Immunoaffinity Column Purification

Note: Do not let column liquid drain dry before step 4.

1. 1. Take out Immunoaffinity Column microcolumn, then take off the top plug and cut it. And then plug it back into the immunoaffinity column.

2. Connect the column with a glass syringe onto the pump flow operation frame, then take off the lower plug. Later, wash it once with 15mL water or phosphate buffer solution (PBS) at pH7.4 at a flow rate of 1-1.5mL/min.

3. Add sample extraction solution at a flow rate of 1-1.5mL/min.

4. Wash it twice with 15mL water or phosphate buffer solution (PBS) at pH 7.4 at a flow rate of 1-1.5mL/min, until 2~3 ml of air passes through the column to ensure that there is no residual liquid in the column.

5. Elute with 1mL methanol at a flow rate of 1mL/min, and use the sample bottle/glass tube to collect the eluent.

6. The collected eluent can be used for detection.

7. Note

8. 1.   1. T2 is harmful to humans, so please wear gloves while operating. All glassware exposed to standard/sample should be soaked overnight with a 5% sodium hypochlorite solution.

   2. Do not use the expired immunoaffinity column.
   3. The Immunoaffinity column should be stored at 2~8℃. Do not freeze.
   4. Please put the immunoaffinity column at room temperature (25℃) for half an hour at least before use.
   5. If the content of T2 in the sample is higher than column capacity, please decrease sample loading volume accordingly.
   6. Make sure that the pH value of the sample extraction solution that passes the column is between 6 and 8. You can adjust it with hydrochloric acid solution or sodium hydroxide solution.
   7. The amount of sample weighed and volume of extraction solution can be adjusted in proportion according to the actual situation. It is recommended to take a 10g sample at a minimum.
      Reference: National standard method: “GB 5009.118-2016 National Food Safety Standard Determination of T-2 Toxin in Food”

HPLC Instrument Measurement Condition

1. 1.       1. Chromatographic Column: C18, 5µm，4.6 mm×250 mm

   2. Mobile Phase: Acetonitrile: Water is 75:25 (V/V)
   3. Flow Rate: 1.0mL/min
   4. Detection Wavelength: Excitation Wavelength 381nm, Emission Wavelength 470nm Sample Loading Quantity: 10µL
   5. Column Temperature: 25℃

1295 Morningside Avenue, Unit 16-18 Scarborough, ON M1B 4Z4 Canada
Phone: 416-261-4865 Fax: 416-261-7879
www.scigiene.com 

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