1. Principle

This test kit is based on the competitive enzyme immunoassay for the detection of Fumonisin B1 in the sample. The coupling antigen is pre-coated on the micro-well stripes. The Fumonisin B1 in the sample and the coupling antigens pre-coated on the micro-well stripes compete for anti-Fumonisin B1 antibodies. After the addition of the enzyme conjugate, the TMB substrate is added for coloration. The optical density (OD) value of the testing sample has a negative correlation with the Fumonisin B1 concentration in the sample. This value is compared to the standard curve and the Fumonisin B1 concentration is subsequently obtained.

2. Technical specifications

Sensitivity : 10 ppb

Detection limit:

Grain, corn, feed.......................................................................... 0.5 ppm

Linear range:

10ng/mL~200ng/mL

Recovery rate:

Grain, corn, feed........................................................................... 90±15%

Precision:
Intra-assay CV<10%;
Inter-assay CV<15%

3. Components

1）   Micro-well strips:96wells(12 strips with 8 removable wells each)

2）   5× standard solution (0.5 mL each): 0 ppb,10 ppb, 50 ppb, 100 ppb,200 ppb

3）   Anti-FB1 monoclonal antibody solution(6ml)............... ready for use directly

4）   Enzyme conjugate (12 mL)......................................... ready for use directly

5）   Substrate solution (12 mL)......................................... ready for use directly

6）   Stop solution (6 mL).................................................. ready for use directly

7）   10× concentrated washing buffer (30 mL)............................... dilute for use

4. Materials required but not provided

1)        Equipments: microplate reader, oscillator,grinder, balance( a reciprocal sensibility of 0.01 g), printer, centrifuge, measuring pipets,.

2)        Material :Graduated cylinder, funnel, flask, rapid qualitative filter paper

3)        Micropipettors: single-channel 20~200 µL, 100~1000 µL; and multi-channel 250 µL;

3)  Reagents:14%Methanol, Sodium chloride (AR),deionized water

5. Sample treatment

Instructions (The following points must be dealt with before the pre-treatment )

1)  Only the disposable tips can be used for the experiments and the tips must be changed when used for different reagents;

2)       Before the experiment, each experimental equipment must be clean and should be re-cleaned if necessary, in order to avoid the contamination that interferes with the experimental results.

Make 30ml 70% Methanol solution:such as

21 mL Methanol + 9 mL deionized water (at volume= 7:3)

Sample treatment:

1)Take 5g smashed sample into centrifuge, add 1.25g Sodium chloride (AR) and 25 mL 70% Methanol solution, shake severely for 3 min.

2) Filter at rapid qualitative filter paper.

3) Take 1mL filtrate, add 9 mL1% NaCl solution for diluting.

4) Take 50 µL for analysis.

Fold of dilution of sample :50