Proposal of the Method for Quantitative Determination of Proteolytic Activity of a Pancreatin Reference Standard

Abstract

Relevance. Pancreatin is a polyenzyme drug with lipolytic, amylolytic and proteolytic activity, and is prescribed as a replacement therapy for exocrine pancreatic insufficiency. Modern requirements for quality control of drugs involve the use of highly specific and highly sensitive methods of analysis using reference standards (RS).For RS certification it is necessary to establish the values of the certifiable characteristic. For enzyme preparations the certifiable characteristic is catalytic activity. In this connection for certification of pancreatin RS it is necessary to establish the value of its catalytic ac-tivity.The analysis of the proteolytic activity of pancreatin consists in performing a proteolysis reaction and quantifying the reaction products. The most common methods used for this purpose are those in which the quantitative content of proteolysis products is determined by measuring the optical density of the solution using tyrosine as a standard. It should be noted that the value of proteolytic activity established in this way is relative, since the measured optical density actually corresponds to a mixture of tyrosine and tryptophan. The absolute method for determining proteolytic activity is the Kjeldahl method for analyzing fragmented peptide nitrogen. Aim. Propose the method for quantitative determination of the proteolytic activity of a pancreatin RS. Material and methods. The object of the study was a pharmaceutical substance pancreatin. Quantitative determination of proteolytic activity in pancreatin substance samples was performed according to the proposed method by determining protein nitrogen by the Kjeldahl method after the en-zymatic reaction. Results. As a result of testing the samples of pancreatin substance using the proposed method of quantitative analysis of proteolytic activity, we ob-tained reliable results, indicating the high sensitivity, accuracy and reliability of this method. Conclusions. The present study proposed the method for quantitative determination of the proteolytic activity of pancreatin substance and evaluated the acceptability of its use in the quality control of a pancreatin RS. Conflict of interest. The authors declare that they have no obvious and potential conflicts of interest related to the publication of this article.