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Protein Digestibility Method

The document describes a method for assessing in vitro protein digestibility by incubating a sample with pepsin and pancreatin over multiple steps. First, the sample is incubated with pepsin in hydrochloric acid for 3 hours. It is then neutralized and incubated with pancreatin in phosphate buffer for 24 additional hours. The digested protein is then precipitated with trichloroacetic acid, centrifuged, and dried before determining nitrogen content to calculate digested versus undigested protein.

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30 views

Protein Digestibility Method

The document describes a method for assessing in vitro protein digestibility by incubating a sample with pepsin and pancreatin over multiple steps. First, the sample is incubated with pepsin in hydrochloric acid for 3 hours. It is then neutralized and incubated with pancreatin in phosphate buffer for 24 additional hours. The digested protein is then precipitated with trichloroacetic acid, centrifuged, and dried before determining nitrogen content to calculate digested versus undigested protein.

Uploaded by

jainrahul2910
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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In-Vitro Protein Digestibility

Protein digestibility (in vitro) was assessed by employing pepsin and pancreatin
following method of Akeson & Stahmann, (1964).
Initially 100 mg of dried and defatted sample was incubated with 1.5 mg pepsin in 15
ml of 0.1 N HCl at 37 C for 3 hrs.
After neutralization with 7.5 ml of 0.2 N sodium hydroxide and addition of 4 mg
pancreatin in 7.5 ml of phosphate buffer with pH 8.0.
The digestion mixture was incubated for an additional 24 hours at 37o C.
Then 5 gm of trichloroacetic acid (TCA) was added to the protein digest.
The protein was precipitated with 10% TCA at the end of experiment.
Finally digested sample was centrifuged at 5000 rpm for 10 min.
Then supernatant was discarded and residue was oven dried at 40-50oC.
The nitrogen contents of the sample and the undigested residue were determined by the
micro-Kjeldahl method (AOAC, 1995).
The digested protein of the sample was calculated by subtracting residual protein
from total protein of the sample.

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