Test-system for estimation of the crosslinking effect of preservative solutions

Purpose of the Study - to develop the technique for evaluation of the crosslinking effect of storage solutions based on analyzing changes in protein electrophoretic spectra and recording the forming intermodular crosslinks. Material and Methods. The crosslinking effect was evaluated with freshly prepared Melnikov-Razvedenkov storage solution containing glycerin and potassium acetate, and the similar storage solution in which the biological material containing human bone, cartilage and muscle tissues was preserved for a long time. Electrophoresis of bovine serum albumin (treated with storage solutions) was made according to Lammle technique in polyacrylamide gel plates with 5-20% acrylamide concentration. Protein lysate of rat's heart muscle was used as marker proteins for the purpose of determining relative molecular weight. Results. The technique developed by the authors on the basis of analyzing changes in protein electrophoretic spectrum allows to reveal the substances in storage solutions which have a crosslinking effect on protein molecules. The change in the relative molecular weight of bovine serum albumin makes it possible to record the presence of formaldehyde and glycerin aldehydes in solutions at the level of the hundredths of a percent. Incubation of bovine serum albumin in the storage solution being in contact with biological material leads to appearing high-molecular-weight fractions evidencing of the formation of inter- and intramolecular crosslinks.

storage solution, bovine serum albumin, formaldehyde, glycerin aldehyde, electrophoresis in polyacrylamide gel