Continuous method to determine the trypsin inhibitor activity in soybean flour

Ezequiel R. Coscueta, Manuela E. Pintado, Guillermo A. Picó, Gastón Knobel, Carlos E. Boschetti, Luciana Pellegrini Malpiedi, Bibiana B. Nerli*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

23 Citations (Scopus)


The determination of trypsin inhibitor (TI) activity is of importance to evaluate the nutritional value of soybean flours. An analytical method, which involves a continuous spectrophotometric rate determination for trypsin activity against the substrate N-benzoyl-DL-arginine p-nitroanilide, is proposed as an alternative to the standard discontinuous assay. Stopping the reaction with acetic acid and a centrifugation/filtration step to decrease turbidity are not required, thus reducing costs and sample preparation time. The TI activity of different flour samples, determined by both assays, demonstrated to be statistically comparable, irrespective of the TI concentration level. The coefficients of variation of the novel method did not exceed 8% at any concentration level. The curves of progress reaction showed a non-linear behavior in samples without TI. A reduction of incubation time from 10 min to 2 min increased the method sensitivity and extended its linear range. A more economical, faster and simpler assay was developed.
Original languageEnglish
Pages (from-to)156-161
Number of pages6
JournalFood Chemistry
Publication statusPublished - 1 Jan 2017


  • Analytical method
  • Antinutritional factors
  • Soybean flour
  • Trypsin inhibitor activity


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