ENCAPSULATION OF PARTIALLY PURIFIED BROMELAIN FROM PINEAPPLE CORES IN ALGINATE-PECTIN BEADS AS A TARGETED ANTIPLATELET AGENT

Objective: Oral administration of bromelain can decrease its bioactivity once it makes contact with stomach acid. Therefore, bromelain was loaded into alginate (Alg), pectin (Pec), and alginate-pectin (AP) beads to control its release into the intestines and avoid degradation. Methods: Crude bromelain was purified by ammonium sulfate precipitation and the dialysis process. In vitro releases and kinetics studies of bromelain-loaded alginate-pectin beads were carried out using an acid and phosphate buffer medium. The beads were characterized using physical analysis, Fourier-Transform Infrared Spectroscopy (FTIR) analysis, and Differential Scanning Calorimetry (DSC) analysis. Results: The dialysis fraction of bromelain has a specific activity of 67.93 U/mg, 3.05 times that of crude bromelain. That fraction was encapsulated in Alg, Pec, and AP beads with a range of encapsulation efficiency around 82.70–91.39%. Bromelain-loaded pectin and AP19 beads were chosen to study in an in vitro release based on their swelling properties and encapsulation efficiency. Bromelain-loaded AP19 beads have lower release than bromelain-loaded pectin beads in both dissolution mediums. The cumulative releases of AP19 are 9.99 and 87.81% in 0.1 N HCl and phosphate buffer medium, respectively. Bromelain-loaded P and AP beads both follow the zero-order kinetics model and the dissolution mechanism of the beads is non-Fickian with a combination of diffusion and erosion. The in vitro antiplatelet activity of dissolution aliquots (20.51 and 18.48%) is lower than its dialysis fraction (56.04%). Conclusion: This in vitro research data shows promising potency for AP as a carrier for oral administration of bromelain as an antiplatelet agent

THE EVALUATION OF ACTIVITY AND STABILITY OF ISOLATED BROMELAIN FROM PINEAPPLE CORES (ANANAS COMOSUS [L.] MERR) AND IN VITRO PENETRATION TEST OF NANOEMULSION TOPICAL BASE

Objective: Oral administration of bromelain as an anti-inflammation therapy still faces several challenges, such as the risk of contact with the gastric fluid and its low absorption rate. Therefore, bromelain isolated from the pineapple core will be formulated as a topical base in a nanoemulsion to increase its stability, activity, and ability to penetrate the skin. Methods: Bromelain was isolated from pineapple core using ammonium sulfate precipitation and dialysis, and then the isolated fraction was loaded into topical nanoemulsion for subsequent evaluation of its characteristics, stability, enzymatic activity, and for in vitro study of penetration using Franz diffusion cell. Results: The highest specific activity of bromelain fractions found in ammonium sulfate concentration is around 20–50%. After being dialyzed, the bromelain fraction showed an increase in specific activity 2.78-fold as compared to crude extract. The characteristics of bromelain nanoemulsion showed a globule size of 21.37±1.8 nm with a polydispersity index (pdI) 0.323±0.049, oil in water (o/w) type, and the type of rheology was plastic flow. The nanoemulsion stability base was observed, and there was no phase separation after centrifugation. Bromelain in nanoemulsion base showed a proteolytic activity of 5.00 U/ml with a protein content of 154.28 mg/l. In vitro penetration studies using Franz diffusion cell for 8 h showed isolated bromelain in nanoemulsion base has a total cumulative number value of 1386.94 μg/cm2 with penetrated velocity/flux (J) of 45.93 μg/cm2 h. Conclusion: The results showed promise for bromelain loaded into nanoemulsion as a vehicle for topically administered therapeutic enzymes

DISSOLUTION KINETICS OF PARTIALLY PURIFIED BROMELAIN FROM PINEAPPLE CORES (ANANAS COMOSUS [L.] MERR.) ENCAPSULATED IN GLUTARALDEHYDE-CROSSLINKED ALGINATE-GUAR GUM

Objective: Bromelain is susceptible to low pH and thus must be encapsulated in glutaraldehyde-crosslinked alginate-guar gum (Alg-GG) hydrogels to avoid bromelain activity degradation in the stomach. Methods: Isolated crude bromelain was purified through ammonium sulfate precipitation, sodium benzoate precipitation, and dialysis. Bromelain-loaded Alg-GG was dissolved in artificial gastric fluid and intestinal environment. Results: The bromelain fractions showed a higher specific activity (U/mg) than the crude enzyme (51.32), as follows ammonium sulfate fraction (267.70±4.67), sodium benzoate fraction (115.63±3.35), and dialysis fraction (332.22). The dialysis fraction was encapsulated in Alg-GG hydrogel containing 0.75% (v/v) glutaraldehyde through the post-loading method. The swelling ratios of the hydrogel are 188.43% at pH 1.2 and 563.83% at pH 7.4. The highest encapsulation efficiency is 72.2%. The maximum bromelain concentration released during dissolution is higher in artificial intestinal environment (1.97 mg/L) than in artificial gastric fluid (0.18 mg/L), and the maximum proteolytic activities are 1.3 and 0.15 U/mL, respectively. Data were incorporated into the zero-order, first-order, Higuchi, and Korsmeyer–Peppas models to determine the kinetics and mechanism of bromelain dissolution. All bromelain concentrations (70, 140, and 210 ppm) follow the Korsmeyer–Peppas model. The dissolution mechanism is a combination of diffusion and erosion. Both the dialysis fraction (56.59%) and the dissolution product (47.45%) showed a good in vitro antiplatelet activity. Conclusion: The present data show the promise of Alg-GG encapsulation as a vehicle for orally administered therapeutic enzymes