Pribolab - IAC-011-1-25PriboFast®黄曲霉素总量免疫亲和柱-1ml-14272

Product  Usage

The  product  is  used to absorb the aflatoxins(B1   B2  G1  G2) in  the   sample  extracting  solution   selectively.    It    plays  a purified   role. The sample  solution  will   be  increased  after using the  immunoaffinity  column. And  it  can  be  detected by different  methods.

Product overview

Aflatoxin  is the secondary  metabolites of aspergillus

flavus  and  parasitic aspergillus fungus.  It  is divided  into

class  1 carcinogen  by the world  health organization's

cancer  research institute,  but it is also a  kind of highly

toxic  highly toxic substances.  It  is widely found  in various foods such  as corn,  peanuts,  rice and wheat and its

by- products. Aflatoxin  B1  is still  highly toxic and

carcinogenic after animal  metabolism, which  is widely

found  in  milk,  blood, tissues  and  other foods. As one  of the three  main causes  of  liver cancer, aflatoxin  is a  major cause of gastric cancer  and  bowel  cancer, which   poses  a great threat to  human  health. There  are  strict  limits  on the amount of aflatoxin in food and feed.

Reaction  principle

This  product  is  based on antigen antibody  reaction.

Aflatoxin  monoclonal antibody  immobilized on the

column  in the gel. The aflatoxin  in the  samples  must  be handled through extraction, filtration  and  dilution.  and then  put the sample extracting  in the aflatoxin

immunoaffinity  column and  make  it flow through the

column slowly. Aflatoxin can combine with the antibody specificity  in the column. Wash the  immunoaffinity

column,  remove other substances that can’t  be

combined.  Elute aflatoxin    with  methanol  and      inject it     to      analytical      instrument     for     testing.

Product  packaging  composition:

Each    box  contains    aflatoxin  total   immunoaffinity columns and a set of corresponding  instructions

IAC  performance:

1 ） Performance  parameters

2） Cross  reaction  rate

Warning and  Precautions

1. Aflatoxin  is  harmful to  human  body, you should wear the glove to  operate.

2. It  is  best to  use the 5%  sodium  hypochlorite solution to soak the  used glass containers and aflatoxin solution one night.

3.   Do  not  use the immunoaffinity column which  is beyond the expiration date.

4.The  immunoaffinity column should  be stored at 2-8  ℃ when  it  is  not  in use, do  not frozen.

5.Before  using, the  immune affinity column should  be restored to  room temperature  (22-25  ℃).

6. Sampling  quantity: according to the actual situation,  it can  be  added and decreased appropriately,  but  not too   little,  and the volume  shall  be changed  according to the proportion

7.   PH: the  pH of the  upper column  liquid should  be

between  6 and 8 .  If  it deviates from this  range, the  pH should  be adjusted with  hydrochloric acid or sodium hydroxide.

Materials  needed  but  not  provided:

Recommended solvents and  buffers:

①  Extraction:  methanol-water  (7:3)

Remove  700 mL  methanol and  300 mL distilled water ②  Extraction: acetonitrile - water  (84:16)

Remove 840 mL acetonitrile and  160 mL distilled water ③  Dilution:

PBS-buffer(10mM):

8 g  NaCl;  1.2 g  Na2HPO4; 0.2 g  KH2PO4; 0.2 g  KCl  (all  p.A. quality),  dissolve  in 990  mL of distilled or  deionized wate r and adjust  pH to 7.4  using  NaOH (1  M) or  HCL (1  M), fill

up to  1000  mL with distilled or deionized water 1% Trition X-100  (or twin-20)  PBS:

10mL Trition X-100  (or twin-20),  diluted with  PBS to  1000 mL.

④  Rinse:  distilled water or deionized water

⑤  Eluent: methanol Sample   processing:

The  maximum error     of  mycotoxin analysis  is the

collection and sampling  process of the sample, so  it  is

important to  ensure the  representative sample collection and sampling.

1  、 Ordinary solid sample

1）Take 5g samples (accurate to 0.01 g)  in 50ml centrifuge

tubes;

2）   Add  20 .0mL acetonitrile - water (84:16) or

methanol-water  (7:3) solution, swirl and  mix well, and

place in the  ultrasonic/vortex oscillator or oscillating  bed for 20min  (or  high speed  homogenous 3min);

3） Centrifuge  10min  under 6000r/min  (or , filter with glass fiber filter  paper to  clarify  after  homogenizing)

4） Get 4  ml filtrate ,add     46  mL  1 % Trition X -  100  (or twain - 20)  PBS  (  It can  be  halved to join when  using

methanol  aqueous  extraction ),  Blending  (such as

turbidity,  please filter to  clarify with glass fiber filter paper)

2 、 Plants oils

1）Take  5g samples (accurate to 0.01 g)  in 50ml centrifuge tubes;

2）Add  20 .0mL acetonitrile -water (84:16)  or

methanol-water  (7:3) solution, swirl and  mix well, and

place in the  ultrasonic/vortex oscillator or oscillating  bed for 20min  (or  high speed  homogenous 3min);

3） Centrifuge  10min  under 6000r/min. Remove the clear liquid for the  next step.

4） Get 4  ml filtrate ,add     46  mL  1 % Trition X -  100  (or twain - 20)  PBS  (It can  be  halved to join when  using

methanol aqueous extraction),  Blending (such as

turbidity,  please filter to  clarify with glass fiber filter paper)

3 、 Soy sauce,  vinegar

1)Take  5g samples  (accurate to 0.01 g)  in 50ml centrifuge tubes;

2 ） Determine with acetonitrile or  methanol to 2 5 ml    (accurate to 0. 1  mL)swirl and  mix well, and  place in the ultrasonic/vortex oscillator or oscillating  bed for 20min (or  high speed  homogenous 3min);

3） Centrifuge  10min  under 6000r/min (or filter with glass fiber filter  paper to clarify after  homogenizing)

4） Get 4  ml filtrate ,add     46  mL  1 % Trition X -  100  (or twain - 20)  PBS  (  It can  be  halved to join when  using

methanol aqueous extraction),  Blending (such as

turbidity,  please filter to  clarify with glass fiber filter paper)

Note:Isotope  diluent  phasechromatography-tandem

mass spectrometry with  reference to GB  5009 . 22-2016 . Operating  procedures： with  reference to GB

5009.22-2016.

1.Connect:  remove the  back  cover of the  immunoaffinity

column  .Cut away with scissors,cover again.

Then  make  it connected to the  30mL  syringe of thePump flow  rack

2. Add Sample:  remove the  above  extraction  liquid to the syringe and open the  lower end of the end cap;

3.Gathering: connect the air  pressure  pump with the

syringe, adjust the  pressure to  make the sample

extracting solution through the  immunoaffinity column slowly.  Recommend  using  immunoaffinity column  by gravity ;

4. Washing:  use 20 mL of distilled water,  clean  it twice with a slight  pressure, discard all effluent, and  make  2-3 ml air through the cylinder;

5.Washing out: to  ensure adequate wash out,  it  is

recommended to take two  steps  of  methanol with 2.0mL

of  methanol, and the gravity will  be the  best.  First, drain the  immune affinity column, add  1.0mL of methanol to

the column  by the gravity. When the solvent  passes

through the column,add  1.0  mL  methanol and  repeat the above steps to  elute, and finally  purge eluent with  a

slight  pressure ,  merge the  eluent, condensed the eluent, dissolve it again with  1  mL  mobile  phase, mix 30s to

dissolve  residue,0 . 2 2 um  micropore filter, prepared for detecting.

Operation flow diagram

Product      Information:

Note: The  Immunoaffinity Column  has  been  passed the internal  quality  control   procedures  and  is  released  for sale.It  is only  used for  laboratory.