Aflatoxin Total (B1, B2, G1, G2) Immunoaffinity Columns

• Express: DHL FedEx
• Samples: Rice Peanut Feed
• Recovery Rate: ≥80%
• Transport Package: Ice Bag with Foam Box
• Specification: 50T
• Trademark: kwinbon
• Model NO.: KH01101Z
• Origin: China
• HS Code: 38229000
• Production Capacity: 5000000000 PCS

Product Description

Aflatoxin total (B1, B2, G1, G2) Immunoaffinity columns

1. Usage

The AFT columns are used by combining with the HPLC, LC-MS, ELISA test kit. It can be quantitative test the AFB1, AFB2, AFG1, AFG2. It is suitable for the cereals, food, Chinese medicine, etc and improves the purity of the samples. The samples after depuration can be tested by different assay way.

1. Test Principle

The aflatoxin immunoaffinity column is based on antigen- antibody-reaction technology. The monoclonal antibody against AFT is linked with the coagulating media in column. AFT in sample is extracted, filtered and diluted. Make the sample extraction solution goes through the AFT column. AFB1, AFB2, AFG1, AFG2 is combined with the antibody separately in column, the washing solution get rid of the impurity not combined. Finally, using the methyl alcohol to elute AFB1, AFB2, AFG1, AFG2. And then test.

1. Equipment and Reagent Not Provided

----Analytical balance
----Pulverizer
----Centrifuge
----Vortex mixer (≥200r/min)
----Measuring cylinder
----50ml glass funnel
----Micropipette : 100-1000ul
----Fast qualitative filter paper
----Glass microfiber filter paper (Whatman 934-AH, diameter: 11cm; aperture: 1.5um)
----20ml glass syringe
Reagent
----Acetonitrile (chromatographic grade)
----Methanol (chromatographic grade)
----Sodium chloride
----Deionized water (or distilled water)

1. Reagents Preparation

Solution 1:  PBS solution
Dissolve the total PBS powder (provided) with the 900ml water. Make the PH value till 7.4 using the HCl or NaOH. Finally make the volume till 1L with the water.
Solution 2:  0.1% tween 20-PBS solution
Dissolve the 0.1% tween 20-PBS powder (provided) with the 900ml water. Make the PH value till 7.4 using the HCl or NaOH. Finally make the volume till 1L with the water.
Solution 3:  1% tween 20-PBS solution
Dissolve the 1% tween 20-PBS powder (provided) with the 900ml water. Make the PH value till 7.4 using the HCl or NaOH. Finally make the volume till 1L with the water.
Solution 4:  5M NaOH solution
Weigh 20g NaOH and dissolve with 100ml water.

1. Sample Preparations

5.1 Cereals, peanut and its products
----Smash the sample.
----Take 25.0±0.01g sample and add 5g NaCl.
----Mix with the 125ml methanol-water solution (70+30, v/v).Shake for 15min.
----Filter with the fast qualitative filter paper, or centrifuge at 4000r/min, 5min.
----Take 15ml filter liquor or supernatant and dilute with 30ml water. Filter with the glass microfiber filter paper till clear.
----Take 15ml filtrate (equal to 1.0g sample) go through the column.

1. 1. Sauce

----Take 50.0±0.01g sample.
----Mix with the methanol-water solution (80+20, v/v) till 100ml. Shake for 15min.
----Filter with the fast qualitative filter paper, or centrifuge at 4000r/min, 5min.
----Take 10ml filter liquor or supernatant and dilute with 40ml water. Filter with the glass microfiber filter paper till clear.
----Take 10ml filtrate (equal to 1.0g sample) go through the column.

 

1. 1. Vinegar

----Take 5.0±0.01g sample and add 1g NaCl.
---- Mix with the PBS solution (solution 1) till 25ml. Shake for 15min.
----Filter with the fast qualitative filter paper, or centrifuge at 4000r/min, 5min.
---- Take 10ml filter liquor or supernatant and dilute with 10ml PBS solution (solution 1). Make the PH value till 7.0 using the 5M NaOH solution. Filter with the glass microfiber filter paper till clear.
----Take 10ml blending filtrate (equal to 1.0g sample) go through the column.

1. 1. Nut products

----Take 10.0±0.01g sample.
----Mix with 40ml acetonitrile -water solution (84+16, v/v). Shake for 15min.
----Filter with the fast qualitative filter paper, or centrifuge at 4000r/min, 5min.
----Take 4ml filter liquor or supernatant and dilute with 46ml 1% tween 20-PBS solution(solution 3). Filter with the glass microfiber filter paper till clear.
----Take 20ml blending filtrate (equal to 0.4g sample) go through the column

1. 1. Chinese medicine, Spice and Tea

----Take 10.0±0.01g sample.
----Mix with 40ml acetonitrile -water solution (84+16, v/v). Shake for 15min.
----Filter with the fast qualitative filter paper, or centrifuge at 4000r/min, 5min.
----Take 6ml filter liquor or supernatant and dilute with 54ml 1% tween 20-PBS solution (solution 3). Filter with the glass microfiber filter paper till clear.
----Take 40ml blending filtrate (equal to 1.0g sample) go through the column

1. Assay process

Notice: avoid the column dry throughout the purification process.
----Take out the upper plug of the 3ml column. Cut down the overhang under the thread of the plug. Inset back into the column. Another side is used after fixed with the 20ml work drum; Take out the upper plug of the 1ml column and shear it diagonally and insert back into the column (It is normal to see small bubbles occasionally in the column. It doesn't influence the usage.)
----Take the above blending filtrate and inject in to the syringe. Adjust the pressure of the syringe so that the solution can pass through the column at a rate of 2s/droplet.
----Using the 20ml water to wash the column (Notice: for the darker sample: sauce, vinegar, Chinese medicine, etc. First, use 10ml 0.1% tween 20-PBS solution to wash. And then use 10ml 10% methanol-water solution wash. Finally use 10ml water to wash). The flow speed is 1s/droplet. Discard effluent till 2-3ml air through the column. Confirm there is no residue liquid in column.
----Add 2.0ml methanol to elute. The flow speed is natural gravity till 2-3ml air through the column. Confirm no residue liquid in column.
----Collect the total eluent and for assay.

1. IAC-HPLC TEST

----Go through the organic phase filter membrane (aperture: 0.22um) and test with HPLC.
----Or dry under the 50ºC with nitrogen . Add mobile phase and mix completely. Go through the organic phase filter membrane (aperture: 0.22um) and test with HPLC according to the GB 5009.22-2016.

1. Attentions

----Aflatoxin is harm to human. Please wear the glove to operate. The glass ware touching the sample should be soaked overnight with 5% sodium hypochlorite.
----Don't use the column out of date.
----Please keep the column in 2-8ºC, don't freeze.
----Before use, the column should be returned to room temperature (25ºC) at least half an hour.
---- The sample amount and liquid volume can be adjusted proportionally according to the physical truth. A minimum of 10g samples is recommended.
----Aflatoxin is sensitive to light. Please keep the sample and extraction solution in dark, cool (2-8ºC)place.

1. Storage

Temperature: 2-8ºC

Shelf life:12months.