Application of proteomic analysis for identification of the pharmaceutical substance pancreatin in developing a reference standard


V.I. Gegechkori, А.А. Shatilina, O.Yu. Shchepochkina, O.V. Panchenko, V.N. Kuzina
Sechenov First Moscow State Medical University (Sechenov University), Trubetskaya str., 8/2, Mosсow, 119991, Russian Federation

Introduction. Pancreatin is the drug of choice for replacement therapy of exocrine pancreatic insufficiency. Pancreatin is a polyenzymatic complex of the porcine pancreas. Taking into account modern international requirements regarding the standardization of drugs for ensuring appropriate quality control are used physical, physico-chemical and biological methods of analysis with high specificity and high sensitivity and involving the use of reference standards (RS). One of the main parts of tests in the procedure of certification of RS, including biological RS, is the verification of structure by various complementary methods, such as mass spectrometry, high-performance liquid chromatography, immunoassay, electrophoresis, etc. Our results proved that proteomic analysis in combination with chromatography-mass spectrometry allows to reliably identify pancreatin and to produce RS based on the studied substance. Objective: identification of pancreatin substance using proteomic analysis combined with chromatography-mass spectrometry in developing a RS. Material and methods. The object of the study was a pharmaceutical substance pancreatin. The structure of the sample was determined by chromato-mass spectrometry after electrophoretic fractionation of proteins in polyacrylamide gel with subsequent enzymatic hydrolysis of individual proteins in the gel. The obtained spectra were analyzed using the database of the National Center for Biotechnology Information (NCBIprot). Results. Amino acid sequences of peptide fragments corresponding to porcine pancreatic enzymes were identified by proteomic analysis of the pancreatin substance sample. The findings confirm that the pharmaceutical substance pancreatin is the sum of porcine pancreatic enzymes. Conclusion. The present study showed the possibility of applying the proteomic analysis for identification of drugs of peptide and protein nature on the example of pancreatin substance in developing a RS.

proteomic analysis
reference standards
chromatomass spectrometry

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