Difference between revisions of "Team:Paris Bettencourt/Protocols/Titration protocols for B2 with HPLC"

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Here, I find  2 different protocols to determine the quantity of B2 using HPLC.</b><br/>
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Here, I find  2 different protocols to determine the quantity of B2 using HPLC.<br/><br/>
  
 
I°) In a 250 ml conical flask, put 5g of food sample and 65 ml of 0.1M HCl.<br/>
 
I°) In a 250 ml conical flask, put 5g of food sample and 65 ml of 0.1M HCl.<br/>
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Add 50ml Beta amylase, 500ml Takadiastase in water (small quantity). Put it at 37°C for 18h.<br/>
 
Add 50ml Beta amylase, 500ml Takadiastase in water (small quantity). Put it at 37°C for 18h.<br/>
 
Dilutate with 125ml water.<br/>
 
Dilutate with 125ml water.<br/>
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Fitred with cellulose paper. And do a 2nd filtration with 0.2µm filter paper.</b><br/>
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Fitred with cellulose paper. And do a 2nd filtration with 0.2µm filter paper.<br/><br/>
  
 
II°) Take Xg of food sample and add 15ml of 0.1M HCl.<br/>
 
II°) Take Xg of food sample and add 15ml of 0.1M HCl.<br/>
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Filtred with Albert No 135.<br/>
 
Filtred with Albert No 135.<br/>
 
Made up to 25ml with milli Q water.<br/>
 
Made up to 25ml with milli Q water.<br/>
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Filtered with 0.2µm nylon filter.</b><br/>
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Filtered with 0.2µm nylon filter.<br/><br/>

Revision as of 18:45, 14 August 2015

Here, I find 2 different protocols to determine the quantity of B2 using HPLC.

I°) In a 250 ml conical flask, put 5g of food sample and 65 ml of 0.1M HCl.
Put it in 100°C for 30 minutes.
Cooling, then adjuste Ph to 4.5 with 2.5M sodium acetate.
Add 50ml Beta amylase, 500ml Takadiastase in water (small quantity). Put it at 37°C for 18h.
Dilutate with 125ml water.
Fitred with cellulose paper. And do a 2nd filtration with 0.2µm filter paper.

II°) Take Xg of food sample and add 15ml of 0.1M HCl.
Incubate at 100°C for 30 minutes.
Cooling, then add 2.5M sodium acetate (to have a Ph4.5) and add 10mg of Takadiastase. Incubate it at 50°C during 3h.
Filtred with Albert No 135.
Made up to 25ml with milli Q water.
Filtered with 0.2µm nylon filter.