Influence of Tableting on Enzymatic Activity of Papain along with Determination of Its Percolation Threshold with Microcrystalline Cellulose

Abstract

The binary mixture tablets of papain and microcrystalline cellulose (MCC), dicalcium phosphate dihydrate (DCP), carrageenan, tragacanth, and agar were prepared by direct compression. Carrageenan, tragacanth, and agar provided maximum protection to enzyme activity compared to MCC and DCP. However, stability studies indicated highest loss of enzyme activity with carrageenan, tragacanth, and agar. Therefore, compression behaviour of different binary mixtures of papain with MCC at different compaction pressures, that is, 40–280 MPa, was studied according to Heckel equation. The compressibility studies of binary mixtures indicated brittle behavior of papain. The application of percolation theory on the relationship between critical density as a function of enzyme activity and mixture composition revealed the presence of percolation threshold for binary mixture. Papain-MCC mixture composition showed significant percolation threshold at 18.48% (w/w) papain loading. Microcrystalline cellulose provided higher protection during stability study. However, higher concentrations of microcrystalline cellulose, probably as dominant particles, do not protect the enzyme with their plastic deformation. Below the percolation threshold, that is, 18.48% (w/w) papain amount in mixture with plastic excipient, activity loss increases strongly because of higher shearing forces during compaction due to system dominance of plastic particles. This mixture range should therefore be avoided to get robust formulation of papain.

Effect of Compaction Pressure on the Enzymatic Activity of Pancreatin in Directly Compressible Formulations

“…Regarding calcium phosphate, two types employed in the study, i.e., DCPA (DI-CAFOS ® A150) and TCP (TRI-CAFOS ® 500), were characterized by high porosity and large specific surface area (23 m 2 /g and 85 m 2 /g, respectively) [ 28 ]. These properties were supposed to facilitate obtaining tablets with adequate mechanical strength, while protecting them from excessive densification, which was reported to be one of the factors responsible for enzyme inactivation [ 14 , 15 ].…”

“…Therefore, the tablets prepared in this study were analyzed for hardness, friability, or disintegration time. Tablet density was also an important consideration, as it can affect enzymatic activity [ 15 ].…”

“…Graf et al showed that the use of micro-crystalline cellulose exerted a stabilizing effect on enzymes in pancreatin tablets [ 13 ]. However, Kuny and Leuenberger as well as Sharma et al noticed that at higher concentrations, MCC can induce high shear forces and cause the enzyme particles to be crushed [ 14 , 15 ]. In their study, Sharma et al linked the loss of model enzyme activity to a decrease in porosity and an associated increase in tablet density under applied compression forces.…”

“…In their study, Sharma et al linked the loss of model enzyme activity to a decrease in porosity and an associated increase in tablet density under applied compression forces. This was explained by the collapse of voids (pores) in compressed powders leading to mechanical damage of protein molecules [ 15 ]. It should be noted that enzyme powders are not identical and differ in terms of their mechanical properties and deformation characteristics.…”

Hydrolysis of Soybean Milk Protein by Papain: Antioxidant, Anti-Angiotensin, Antigenic and Digestibility Perspectives

“…A UV-Vis spectrophotometer (Thermo Scientific TM , Waltham, MA, USA) was used for that purpose. One USP unit of papain activity represents the equivalent release of 1 mg of tyrosine from a casein substrate solution [ 79 ].…”