Utilization of Industrial Rosa damascena Mill. By-products and Cocoa Pod Husks as Natural Preservatives in Muffins

Cocoa Pod Husks (CPH) and by-product from supercritical CO2 extracted Rosa damascena Mill. (RDCO2) were used as biopreservatives in muffins. Both by-products were rich source of polyphenols: 28.3 ± 0.6 mg/g Dry Weight (DW) and 17.9 ± 0.7 mg/g DW RDCO2 and CPH, respectively, and exhibited potent antioxidant capacity: 449.1 ± 8.5 µmol Trolox Equivalents (TE)/g DW (by ORAC method) and 58.9 ± 2.1 µmol Gallic Acid Equivalents (GAE)/g DW (by HORAC method) for the RDCO2, and 373.8 ± 9.0 µmol TE/g DW (by ORAC) and 36.8 ± 3.8 µmol GAE/g DW (by HORAC) for the CPH. RDCO2 extracts successfully inhibited development of several important pathogenic and saprophytic microorganisms causing microbial spoilage of food systems. The control muffins were good for consumption up to the 17th day, while the products supplemented with RDCO2 and CPH: until 20th day of storage at 22 ± 0.5 °C. The amount of dietary fibers in muffins supplemented with both by-products increased 3 times (8.57 ± 0.12 %) compared to control (2.91 ± 0.12 %) and the polyphenolic compounds increased 2.5 times (from 50.0 ± 0.3 for the control to 185.9 ± 0.6 mg/g DW). For the first time by-product of supercritical CO2 extraction of Rosa damascena Mill. was characterized and used as natural and cheap biopreservative.

Screening Cellulolytic Fungi Isolated From Malaysia Cocoa Pod Husk and Its Culture Conditions for Cellulases Production

The aim of this study was to screen few fungal isolates from local cocoa pod husks (CPH) which able to secrete cellulases. The isolates were plated on carboxymethyl cellulose (CMC) agar plates which then incubated for two days at 28ºC. Then, these plates were stained with congo red dye for 0.5-1 h followed by destaining with 1 M NaCl solution for 15-20 minutes to observe its cellulolytic activity. One isolates which exposed the largest cellulolytic zone on CMC agar plate was selected for further study. In this study, culture conditions with respect to pH, incubation time, amount of substrate (CPH) and temperature were screened using Design expert @version 8.0 by employing two-level factorial design. The selected fungus isolate was cultured in shake flask at 37°C with agitation of 200 rpm for 5 days in incubator shaker. During fermentation period, samples were collected every day for fungal-cellulases activity of filter paper activity (FPase) and carboxymethyl cellulase (CMCase) activity. Analysis of variance (ANOVA) of this study showed that the most significant parameters that affects the production of cellulases from the selected fungi isolates were the amount of substrate (CPH) used followed by the interaction of amount of substrate with pH (p< 0.05). It showed that the cellulases activity was high when the pH 9 with more amount of substrate used. However, it was observed that less significant changes of celllulases activity when different amount of substrate was used at same pH of 3. Based on the microscopic observation of isolate, it morphology was closed to Rhizopus sp.. In conclusion, it is suggested to optimize the selected culture parameters obtained in this study in order to maximize the activity of cellulases from the selected isolates.

A Pharmacokinetic Evaluation of a Pectin-Based Oral Multiparticulate Matrix Carrier of Carbamazepine

Background. Carbamazepine is a drug used in the treatment of neurological disorders such as epilepsy. However, due to its erratic absorption, oral bioavailability is often poor. There is, therefore, the need to develop alternative formulations for carbamazepine with better pharmacokinetic characteristics. Aim. The aim of this study was to formulate an oral modified-release multiparticulate matrix of carbamazepine from cocoa pod husk (CPH) pectin and evaluate the pharmacokinetic profile of this formulation using in vitro and in vivo models. Methods. CPH pectin was extracted from cocoa pod husks with hot aqueous and citric acid solutions. Oral multiparticulate carbamazepine matrices were formulated from CPH pectin cross-linked with calcium. The formulation was evaluated for carbamazepine content and release profile in vitro. For in vivo pharmacokinetic profile estimation, rats were put into 4 groups of 5 animals each to receive carbamazepine multiparticulate matrix formulations A and B, carbamazepine powder, and Tegretol CR®. Animals in each group received 200 mg/kg of each drug via the oral route. Maximum plasma concentration C max , area under the concentration-time curve (AUC), elimination rate constant K e , and terminal half-life t 1 / 2 of the formulations were estimated by noncompartmental analysis. Results. The pectin extraction from fresh cocoa pod husks using hot aqueous and citric acid solutions gave pectin yields of 9.63% and 11.54%, respectively. The drug content of carbamazepine in CPH pectin formulations A and B was 95% and 96%, respectively. There was controlled and sustained release of carbamazepine for both formulations A and B in vitro. AUC0⟶36 (176.20 ± 7.97 µg.h/mL), C max (8.45 ± 0.71 μg/mL), T max (12 ± 1.28 h), and t 1 / 2 (13.75 ± 3.28 h) of formulation A showed a moderately enhanced and comparable pharmacokinetic profile to Tegretol CR® (AUC0⟶36: 155 ± 7.15 µg.h/mL, C max : 8.24 ± 0.45 μg/mL, T max : 8.0 ± 2.23 h, and t 1 / 2 : 13.51 ± 2.87 h). Conclusion. Findings from the study suggest that formulations of CPH pectin had the potential to control and maintain therapeutic concentrations of carbamazepine in circulation over a period of time in the rat model.

Blood Pressure Regulating and Antioxidant Potentials of Theobroma Cacao Pod Husk Protein Hydrolysates

Background: Agrowastes like Theobroma cacao (Cocoa) pod husk can be used to prepare bioactive peptides with various bio-functionalities. Objectives: This study aimed to investigate antioxidant and angiotensin converting enzyme I (ACE) inhibitory peptides contained in Theobroma cacao (cocoa) pod husks – an agro-waste. Methods: Protein isolated from cocoa pod husk was enzymatically digested with alcalase, pepsin, and trypsin. ACE inhibition, kinetics of ACE inhibition, and antioxidant properties of the cocoa pod husks hydrolysates were evaluated in vitro. Results: Trypsin and alcalase hydrolysates displayed higher peptide yields (63.1% and 61.2%) than pepsin hydrolysate (61.2%). However, no significant difference (P>0.05) was observed in the degree of hydrolysis (DH) of the three proteases on cocoa pod husk protein. Methionine, lysine, and cysteine were the amino acid residues presented in cocoa pod husk hydrolysates. A concentration-dependent ACE inhibition by cocoa pod husk hydrolysates was observed. The highest ACE inhibitions of 84.4%, 81.5%, and 73.5% were obtained at 2.0 mg/mL of pepsin, trypsin, and alcalase hydrolysates, respectively, with the minimum IC50 value of 0.36 mg/mL obtained for trypsin hydrolysate. An uncompetitive and mixed-type inhibition was obtained from double reciprocal plots of alcalase and pepsin as well as trypsin cocoa pod husk protein hydrolysates. The Ki values of ACE inhibition for pepsin, trypsin, and alcalase hydrolysates were 3.05, 2.19, and 3.57 mg/mL, respectively. A concentration-dependent increase in the scavenging of 2,2-diphenyl-1-picrylhydrazyl and superoxide radicals as well as ferric reducing antioxidant power were recorded for the cocoa pod husk hydrolysates. Conclusion: Trypsin and alcalase cocoa pod husk protein hydrolysates could be an effective source of a natural ACE inhibitor and antioxidant.

Characteristics of Pectin Extracted from Cocoa Pod Husks

Cocoa plant husks is one of the source of pectin. Pectin in the food industry is used as thickener, gel agent and stabilizer. The aim of this study was to determine the characteristics of the pectin from cocoa pod husks with variation in temperature and extraction times. In this study the extraction of pectin from cocoa pod husk was carried out with hydrochloric acid solvent, with 40, 60 and 80 minutes extraction times at 65oC, 80oC, and 95oC. The results showed that the extraction temperature (65oC, 80oC, and 95oC) and extraction time (40, 60, and 80 minutes) had no significant effect on metoxyl content, galacturonat acid, equivalent weight, and pectin content, but the extraction time had a significant effect on the acetyl number and the degree of esterification. The highest yield of pectin was obtained at extraction temperature of 80oC for 60 minutes. The pectin content in cocoa pod husk is larger than the content in banana skin. The characteristics of cocoa pod husk pectin powder are brown, with moisture content 10.56-11.96%, ash content 6.82-8.97%, methoxyl content 3.51-4.86%, galacturonic acid content 41.38-88.40%, esterification degree 10.76-19.96%, acetyl number 14.55-20.90%, equivalent numbers 663.83-1549.22, pectin content 9.52-19.51 and yield 5.55- 7.70%. IR Spectrum showed that there was no difference in the functional groups between standard, commercial, and pectin extracted at different temperatures.

How nutrient rich are decaying cocoa pod husks? The kinetics of nutrient leaching

Aim Recycling of cocoa pod husks has potential to contribute to mineral nutrition of cocoa. Yet little is known of the nutrient content and nutrient release patterns from the husks. The potassium (K) rich husks are usually left in heaps in cocoa plantations in Africa. We aimed to understand and quantify release patterns of K and other nutrients from husks under varying rainfall regimes and assessed the effects of partial decomposition and inundation on nutrient leaching rates. Methods We incubated chunks of cocoa pod husks to assess decomposition rates and we measured nutrient leaching rates from two sets of husk chunks: one set was placed in tubes that were submitted to simulated scheduled rainfall events while the second set was continuously inundated in beakers. Results Decomposition of husks followed a second-order exponential curve (k: 0.09 day−1; ageing constant: 0.43). Nutrient losses recorded within 25 days were larger and more variable for K (33%) than for other macronutrients released in this order: Mg > Ca ≈ P > N (less than 15%). Potassium leaching was mainly driven by rainfall frequency (P < 0.05) and reinforced by intense rainfall, especially at lower frequency. Under water-saturated conditions, 11% of K was leached out within 48 h from fresh husks compared with 92% from partially decayed husks. Conclusion Some initial decomposition of cocoa pod husks is required to expose K to intense leaching. As decomposition progresses, abundant K losses are to be expected under frequent and/or intense rainfall events.

Enzymatic Extraction and Characterization of Pectin from Cocoa Pod Husks (Theobroma cacao L.) Using Celluclast® 1.5 L

Cocoa pod husks are a waste generated during the processing of cocoa beans. We aimed to explore the enzymatic extraction of pectin using cellulases. The extraction process was optimized using a central composite design (CCD) and analyzed by response surface methodology (RSM). The parameters optimized were feedstock concentration (%), enzyme dosage (µL/g), and time (h). Three dependent variables were studied: pectin yield (g/100 g dry husk) (R2 = 97.02), galacturonic acid content (g/100 g pectin) (R2 = 96.90), and galacturonic acid yield (g/100 g feedstock) (R2 = 95.35). The optimal parameters were 6.0% feedstock concentration, 40 µL g−1 of enzyme, and 18.54 h, conditions that produced experimentally a pectin yield of 10.20 g/100 g feedstock, 52.06 g galacturonic acid/100 g pectin, and a yield 5.31 g galacturonic acid/100 g feedstock. Using the chemical extraction method, a yield of 8.08 g pectin/100 g feedstock and a galacturonic acid content of 60.97 g/100 g pectin were obtained. Using assisted sonication, a pectin yield of 8.28 g/100 g feedstock and a galacturonic acid content of 42.77 g/100 g pectin were obtained. Enzymatically optimized pectin has rheological and physicochemical features typical of this biomaterial, which provides an interesting alternative for the valorization of cocoa husks.