Pod Husk Extract Research Articles

Toxicity of methanolic extract from cocoa pod husk (Theobroma cacao L) using brine shrimp lethality test (BSLT)

Abstract Indonesia is one of the countries that produces a large amount of Theobroma cacao L. in the world. The extract of water, ethyl acetate, chloroform, and n-hexane of cocoa pod husk contains several chemical constituents that exhibit potential biological activities. This study aimed to determine the toxicity of cocoa (Theobroma cacao L.) pod husk extract. Cocoa pod husk (6 kg) was first macerated with methanol and then extracted using n-hexane to obtain n-hexane extract (TCH) weighing 68.81 g (40.91%). The methanol extract was further extracted with ethyl acetate to obtain 17.98 g (10.69%) ethyl acetate extract. The methanol extract thus obtained was 76.38 g (45.41%). The methanol extract was further extracted using acetone to give 11.91 g (15.59%) acetone extract (TCA) and 45.33 g (59.35%) methanol extract (TCM). Theobroma cacao Acetone extract (TCA) was further isolated by gravity chromatography column and 11 fractions (TCA 1-TCA 11) were obtained. TCA 3 was re-chromatographed using an eluent system of n-hexane: ethyl acetate (8.5:1.5). The toxicity of the TCA extract and its combined fractions (TCA 1-TCA 11) on Artemia salina Leach larvae was assessed. TCA fraction 3 was found to be toxic with an LC50 value of 136.3 ppm.

The Effect of Cocoa Pod Husk (Theobroma cacao L.) Extract on Increasing Gingival Fibroblast and Collagen Density in Wistar Rats Induced by Porphyromonas gingivalis

Periodontal disease is an inflammatory disease of the oral cavity which characterized by inflammation of the gingiva. Theobroma cacao L. is a herbal plant that has antibacterial and anti-inflammatory effects, which is starting to be widely researched as an alternative therapy for inflammation in the oral cavity. The aim of this study was to examined the effect of the ethanol extract gel of cocoa pod husk (Theobroma cacao L.) on increasing the number of fibroblasts and the collagen fiber density in the gingiva of rats that have been induced by Phorphyromonas gingivalis. The sample used 24 male Wistar rats divided into 3 groups. Each group was induced by porphyromonas gingivalis every 3 days for 14 days then anointed with placebo gel (negative control group), metronidazole gel (positive control group), and cocoa pod ethanol extract gel (CPH gel) for 7 and 14 days. Gingival tissue was taken after euthanasia, after which histological preparations were made, and fibroblasts were observed under a microscope and analyzed using quantitative pathology 0.3.2 software, while for collagen density analized using Adobe Photoshop CS6 software. The results showed there was a significant difference of the number of fibroblasts between each group (p<0.05). however there was no significant difference between positive control and cocoa pod husk extract at day 14 (p > 0.05).The treatment group on day 7 and 14 was higher than the other groups. It was conclude that the ethanol extract of cacao pod husk (Theobroma cacao L.) potential to increase the number of fibroblasts and collagen fibers density in gingival tissue of periodontitis rats.

Novel combination of nanostructured calcium hydroxide and natural materials: Formulation and characterization.

Calcium hydroxide is used as a reparative dentin inducer. The combination of cocoa pod husk and calcium hydroxide has also been studied as a reparative dentin inducer, whereas anchovy was identified as a good calcium source. The aim of the study was to investigate the optimal ratio of a nano-combination of calcium hydroxide, cocoa pod husk extract and anchovy powder as a reparative dentin inducer by analyzing their material, antioxidant and cytotoxic properties. The ratios of the 3 elements were established and their performance was compared in terms of viscosity, setting time, pH, and solubility rate. The optimal ratio was then examined for its antioxidant capacity using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. Additionally, its cytotoxicity against human dental pulp stem cells (hDPSCs) was evaluated through the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. A ratio of 1:1:1 of calcium hydroxide, cocoa pod husk extract and anchovy powder was identified as the optimal ratio. It exhibited the greatest flowability, the shortest final setting time, had a pH range of 8.5-10.5, and the second highest solubility rate in comparison with other ratios. Moreover, it demonstrated active radical scavenging activity and stimulated the proliferation of hDPSCs. Based on the material, antioxidant and cytotoxic properties, a ratio of 1:1:1 of nano-calcium hydroxide, cocoa pod husk extract and anchovy powder was determined to be optimal for use as a reparative dentin inducer.

Comparative analysis of total flavonoid, total phenolic content, and antioxidant activity of Theobroma cacao Pod Husk extract and its derived fractions

Secondary metabolites from plants are known to be promising sources of raw materials for drugs in terms of safety. This research focuses on cacao pod husk as a candidate for discovering natural antioxidant compounds. The stages in this study include the extraction of cacao pod husk using methanol as a solvent, followed by liquid-liquid fractionation to produce ethyl acetate and acetone fractions. Subsequently, total flavonoid and phenolic content, DPPH antioxidant activity tests, and GCMS metabolite analysis were conducted. The research results indicate that the ethyl acetate fraction has the highest total flavonoid and phenolic content compared to methanol extract and acetone fraction. The methanol extract has the highest antioxidant activity with an IC50 of 0.0736 mg/mL. The identified metabolites belong to the groups of polyphenols, flavonoids, steroids, and their derivatives, as well as some volatile compounds such as hexadecanoic acid methyl ester. Compounds with potential antioxidant properties include Eugenol, Myristicine, Piperine, and Theobromine.

Low fat and high protein meat analogue of cowpea (Vigna unguiculata) with stabilizer cocoa pod husk extract (Theobroma cacao L.)

Meat analogue is a plant-based protein product with nutritional and sensory qualities similar to conventional meat. Meat analogue is usually made from wheat flour. People with gluten intolerance can not consume wheat flour. Gluten-free meat analogue is appropriate for such people. In this research, gluten-free meat analogue was made from cowpea flour and cocoa pod husk extract, in three different formulations. Cocoa pod husk extract serves as stabilizer and sticking agent. The formulation was based on the ratio of the main ingredients. The ratios were 98.5:1.5 (A), 99.0:1.0 (B), and 99.5:0.5 (C). The control meat analogue was made from cowpeas flour and commercial pectin with the ratio of 98.5: 1.5. The parameter of quality was the physicochemical aspect including moisture content, ash content, fat content, protein content, carbohydrate content, fibre content, amino acid quality, and in vitro protein digestibility. In addition, the microbiological, texture, and sensory analysis were also conducted. Meat analogue B had 41.56% water content, 4.71% ash content, 3.88% fat content, 19.16% protein content, 8.75% soluble fibre content, 10.21% insoluble fibre content, protein digestibility 65.94%, total plate count of 4.40×102 CFU/g and yeast and mould count of 2.27×102 CFU/g. It is considered the best formulation among the meat analogues.

Analysis of antioxidant and antibacterial activity of cocoa pod husk extract (Theobroma cacao L.)

Background: Theobroma cacao bean manufacturing worldwide includes undesired byproducts such as cocoa pod husks. Cocoa pods contain a variety of beneficial chemicals, including polyphenols. Polyphenols have a vital function in the treatment of many illnesses and infections of the oral cavity. This is due to their vital qualities in the oral cavity, including anti-inflammatory, antibacterial, and antioxidant actions. Purpose: The goal of this study was to look at the antioxidant content and antibacterial activity of cocoa pod husk extract (Theobroma cacao L.) against Phorphyromonas gingivalis (P. gingivalis) and Streptococcus mutans (S. mutans). Methods: Cocoa pod husk extract was produced using an ultrasonic homogenizer and 70% ethanol. Thin layer chromatography and the Folin–Ciocalteu test were used to determine the phytochemical content and total phenolic content of the extract. The 2,2-diphenyl-1-picryl-hydrazyl-hydrate technique was used to measure antioxidant activity. Minimum inhibitory concentration (MIC) tests were used to measure the antibacterial activity of ethanolic extract at concentrations of 1, 4, 8, 16, 32, and 64 mg/ml using a deep-well broth microdilution technique. Results: The presence of alkaloids, flavonoids, tannins, saponins, and triterpenoids was discovered in the cocoa pod husk extract. Conclusion: The antioxidant activity of the extract was significant (IC50 = 62 ppm), and the MIC of P. gingivalis and S. mutans was 16 mg/ml and 8 mg/ml, respectively.

Effect of Cocoa Pod Husk Proanthocyanidins Extract on Cyclooxigenase-2 (COX-2) Expression in Gingival Crevicular Fluid of Periodontitis Rats

Inflammation of periodontal tissue due to Porphyromonas gingivalis which invades the gingival tissue due to the activity of virulence factors. This causes the host immune system changes and increases COX-2 production in periodontal tissue so resulting excessive tissue destruction. The aim of this study was to analyze the potency of proanthocyanidins extract of cacao pod husk to inhibit COX-2 expression in rat gingival sulcus fluid. 3 groups in this study, namely negative control, positive control, and treatment. In all groups, on the 0 day, gingival sulcus fluid was taken, then the rats were induced by Porphyromonas gingivalis once every 3 days for 2 weeks; then in the negative control group a placebo gel was applied and the treatment group was applied proanthocyanidins extract gel of cacao pod husk extract every day for 7, 14, and 28 days and serial gingival crevicular fluid collection was carried out on days 7, 14, and 28. The expression of COX-2 in the gingival crevicular fluid was observed using the ELISA method. The statistical test used was Anova. There were a significant difference in COX-2 expressions (p <0.05). There was a decrease in the expression of COX-2 in rat gingival sulcus fluid induced by Porphyromonas gingivalis and given 10% cacao pod husk extract. Bioactive compounds in cocoa pod husk extract potential to reduce COX-2 expression in periodontitis rats. Article Details

Aktivitas Antioksidan Ekstrak Kulit Buah Kakao (Theobroma cacao L.) Pada Perlakuan Suhu dan Lama Penyimpanan

This research was conducted to determine effect of temperature and storage time on antioxidant activity of cocoa pod husk extract (Theobroma cacao L.) and to obtain the best combination of storage temperature and storage time that can maintain the antioxidant activity of the extract. The experimental design used in this research was a factorial randomized block design, which are grouped into 2 groups based on the implementation time and consisted of two factors. The first factor was the storage temperature which consisted of 3 level that namely room temperature (27 ±2)?, cold temperature (5 ±2)? and freezing temperature (-10 ±2)?. The second factor was the storage time which consisted of 4 level namely 1, 2, 3, and 4 weeks. The data were analyzed by analysis of variance and continued with Tukey test. The results showed that the temperature and storage time had a very significant effect on total phenol, total flavonoid and antioxidant capacity of cocoa pod husk extract. Interactions between treatments had a very significant effect on total phenol, total flavonoid and antioxidant capacity of cocoa pod husk extract. The best treatment showed that freezing storage (-10 ±2)? for 1 week can maintain total phenol was 12.32 ± 0.08 mg GAE/g extract, total flavonoid was 4.07 ± 0.09 mg QE/g extract, antioxidant capacity was 6.50 ± 0.02 mg GAEAC/g extract.

Effects of Cocoa Pod Husk Extract (Theobroma Cacao L.) on Alveolar Bone in Experimental Periodontitis Rats

Periodontitis is a destructive inflammatory disease of the periodontal tissues with a high prevalence in the world. Phorphyromonas gingivalis as one of the main bacteria causing periodontitis produces virulence factors that trigger immune cells to produce proinflammatory cytokines that can reduce the number of osteoblasts, thereby triggering alveolar bone resorption. Osteoblast cell differentiation is mediated by a number of growth factors such as cytokines and bone morphogenic proteins (BMPs). SRP mechanical therapy is sometimes not optimal and Metronidazole as an effective antibiotic against periodontitis has been reported to have several adverse effects. Utilization of cocoa pod waste (Theobroma cacao L) with its polyphenol content has antibacterial, antioxidant and anti-inflammatory effects. This study aims to determine the effect of ethanol extract gel from cocoa pod husk extract (Theobroma cacao L.) on the number of osteoblasts and osteoclasts, as well as the expression of BMP-2 in the alveolar bone of periodontitis-model rats. This laboratory experimental study used 24 rats which were divided into negative control group (CMC-Na gel), positive control group (metronidazole gel), and treatment group (100 mg/mL cocoa pod husk extract gel). 24 Wistar rats were divided into 3 groups which were observed on days 7 and 14. The number of osteoblasts and osteoclasts of alveolar bone were observed by hematoxillin eosin staining and BMP-2 expression by immunohistochemistry staining. One-way ANOVA test showed that there was a significant difference (p < 0.05) between groups. The LSD test average of BMP-2 expression in the treatment group on the day 7 and day 14 was higher than the negative control group (p < 0.05). The conclusion of the study was that giving 100 mg/mL of cacao pod husk extract gel (Theobroma cacao L.) increased the number of osteoblasts and increased the expression of BMP-2 in the alveolar bone of periodontitis rats. HIGHLIGHTS Cocoa pod husk extract gel (Theobroma cacao) 100 mg/mL increased the ratio of osteoblasts to osteoclasts and increased BMP-2 expression in the alveolar bone of periodontitis rats The histological staining with IHC was analyzed using Quantitative Pathology 0.3.2 software The cocoa pod husk (Theobroma Cacao L.) extract gel effectively triggers and increased BMP-2 which stimulates bone formation in periodontitis rat models GRAPHICAL ABSTRACT

The Effect of Adding Cinnamon Bark Oil to the Tyrosinase Inhibitory Activity of Emulgel Containing Cocoa Pod Husk Extract

Our previous study showed that cocoa pod husk extract has tyrosinase inhibitory properties and the potential to prevent skin hyperpigmentation. Another natural source that is known to have tyrosinase inhibitory properties is cinnamon bark oil. This paper aims to analyze the tyrosinase inhibitory properties of cinnamon bark oil through molecular docking. It also aims to determine the effect of adding cinnamon bark oil to the tyrosinase inhibitory properties of emulgel containing cocoa pod husk extract. The constituents of cinnamon bark oil were determined using gas chromatography-mass spectrometry. The molecular docking was conducted using autodock. The emulgels were prepared by adding 2% of cocoa pod husk extract with and without the addition of cinnamon bark oil (1%). Tyrosinase inhibitory properties were analyzed using a colorimetric enzymatic assay and the dopachrome method. The GCMS result showed the cinnamon bark oil containing 53.37% cinnamaldehyde. The in-silico study showed cinnamaldehyde properties as a tyrosinase inhibitor, since it can bind on the active site of the enzyme with free binding energy at -4.88 kcal/mol. The addition of cinnamon bark oil (1%) to the emulgel preparation increased the tyrosinase inhibitory activity by 63.33% based on in vitro study.
 Keywords: emulgel, cocoa pod husk extract, cinnamon bar oil, tyrosinase inhibitor

The immunostimulatory effects of Theobroma cacao L. pod husk extract via injection and dietary administrations on Macrobrachium rosenbergii and its resistance against Lactococcus garvieae

The extract from Theobroma cacao L, pod husk served as the immunostimulant to enhance the immunity and resistance against Lactococcus garvieae of Macrobrachium rosenbergii. In this study, we employed the injection method and dietary administration method to determine the effect of cocoa pod husk (CPH) on M. rosenbergii. The non-specific immune parameters and disease resistance were evaluated after the prawn injected with 1 μg prawn−1 CPH extract (C1), 3 μg prawn−1 CPH extract (C3), and 5 μg prawn−1 CPH extract (C5) for 1, 3, and 7 days. The results showed a significant increase of total haemocyte count (THC), differential haemocyte count (DHC), phenoloxidase (PO) activity, respiratory bursts (RBs), and phagocytic activity and clearance efficiency to L. garvieae. The non-specific immune parameters, physiological parameters, and disease resistance and growth performance were evaluated after the prawn fed with 1 g kg−1 CPH extract diet (CD1), 3 g kg−1 CPH extract diet (CD3) and 5 g kg−1 CPH extract diet (CD5). The results showed a significant increase in all immune parameters and showed a significant decrease in physiological parameters. No significant difference was observed in growth performance of prawn fed with the CPH containing diet. Both injection and dietary method showed a significant increase in disease resistance against to L. garvieae. We therefore recommend that CPH extract can be used as a immunostimulant for prawn by dietary administration to regulate immune responses, and carbohydrate metabolism lead to enhance resistance against pathogen.

THE CYTOTOXIC AND PROLIFERATIVE ACTIVITY OF COCOA POD HUSK EXTRACT (THEOBROMA CACAO L.) ON PERIODONTAL LIGAMENT FIBROBLASTS

Background: Theobroma cacao was considered agricultural wastes. Cocoa pod husk contain various bioactive compounds as natural antioxidants,antibacterial and anti-inflammatory. A good biomaterial must be non-toxic and have no detrimental effect on the biological environment. The aim of research was to determine the cytotoxicity and proliferative activity of cocoa pod husk extract on primary cells of periodontal ligament fibroblasts. Method: This experimental laboratory used primary cell culture of periodontal ligament fibroblasts. The cytotoxicity and proliferative activity were analyzed by MTT assay [2-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide]. Extraction of cacao pods were obtained using the ultrasonic homogenizer method with ethanol 70%. The cytotoxicity test was observed for 24 hours and the doubling time proliferative test was observed for 24, 48, and 72 hours. Result: Cocoa pod husk extract showed there were no cytotoxic effects with viability of primary cells of periodontal ligament fibroblasts at the concentration of 1.56 pg/ml, 3.125 pg/ml, 6.25 pg/ml, 12.5 pg/ml, 25 pg/ml, 50 pg/ml, and 100 pg/ml. The proliferation test showed increasing of fibroblasts at 24 and 48 hours. Kruskal Wallis analysis obtained a significance value (p<0.05) Conclusion: Cytotoxicity and proliferation assays of Cocoa pod husk extract increased cell viability and cell proliferation of periodontal ligament fibroblast.

The Effect of Chitosan-Gelfoam Cacao Pod Husk on Wound Epithelial Thickness in the Post-Extraction Tooth with Anticoagulant Therapy

Tooth extraction is removing a tooth from its socket when it cannot be restored. Tooth extraction performed in patients on anticoagulant therapy can increase the risk of excessive bleeding. Therefore, a local hemostatic agent is needed to accelerate the hemostasis process and reduce the risk of tooth extraction complications in anticoagulant users. This study aims to determine the effect of chitosan-gelfoam extract of cacao pod husk to increase epithelial thickness in the post-extraction socket of male Wistar rats with anticoagulant therapy. This research used quantitative data collection techniques on 24 male Wistar rats who were given anticoagulant therapy. It was divided into eight groups: negative control group H+3, negative control group H+7, positive control with oral tranexamic acid therapy H+3, positive control with oral tranexamic acid therapy H+7, treatment 1 with 1 ml chitosan-gelfoam cacao pod husk extract therapy H+3, treatment 1 with 1 ml chitosan-gelfoam cacao pod husk H+7, treatment 2 with 10 ml chitosan-gelfoam cacao pod husk H+3, and treatment 2 with 10 ml chitosan-gelfoam cacao pod husk H+7. The test animals were decapitated on the 3rd and 7th day. The histology preparations, observations, and statistical tests were then carried out. The statistical tests showed that chitosan gelfoam extract of cacao pod husk with doses of 1.6% and 15% extract of cacao pod husk was able to replace the effectiveness of tranexamic acid. The 15% dose had better significance than the 1.6% dose. This research is expected to contribute to the farmers’ welfare and the cacao industry by converting cacao pod waste into helpful medicine.

Konsentrasi Ekstrak Etanol Kulit Buah Kakao (Theobroma cacao L.) dan Suhu Pemanasan terhadap Karakteristik Krim Body Scrub

Cocoa pod husk extract can be used as a source of antioxidants in cosmetic preparations, namely cream body scrub. Cream body scrub is a type of cosmetic cleanser that is used to clean dead skin cells and nourish the skin. This study aims to (1) know the effect of the concentration of ethanol extract of cocoa pod and heating temperature on the characteristics of cream body scrub and to (2) determine the concentration of ethanolic extract of cocoa pod and the best heating temperature to produce cream body scrub. This study used a factorial randomized block design with two factors. The first factor is the concentration of the extract which consists of 3 levels, namely 1%, 2%, and 3%. The second factor is the heating temperature which consists of 2 levels, namely 65±2°C and 75±2°C. The results showed that the concentration of the extract and the heating temperature and their interactions affected the antioxidant capacity, antioxidant activity, pH, viscosity, spreadability, color, aroma and overall acceptance. Extract concentration of 3% and heating temperature of 65±2°C is the best treatment to produce cream body scrub with antioxidant capacity characteristics of 85.02±0.22 mg GAEAC/g, antioxidant activity 98.48±0.82 ppm (strong antioxidant), pH 6.66±0.02, viscosity 34200±200 cp, spreadability 4.78±0.10 cm, sensory evaluation: color 4.35±0.48(light brown to light brown), aroma 6.25 ±0.71 (fairly typical of cocoa to typical of cocoa) and overall acceptance of 6.25±0.71(like to very like).
 Keywords: cocoa pod husk, concentration, temperature, characteristics, cream body scrub.

Pengaruh Konsentrasi Etanol dan Waktu Ekstraksi menggunakan Metode Microwave Assisted Extraction (MAE) terhadap Aktivitas Antioksidan Ekstrak Kulit Buah Kakao

This research was conducted to determine effect of ethanol concentration and extraction time on antioxidant activity of cocoa pod husk extract and to obtain ethanol concentration and extraction time using Microwave Assisted Extraction (MAE) that can produce extract with the highest antioxidant capacity. The experimental design used in this research was a factorial randomized block design, which consisted of two factors. The first factor was the concentration of ethanol which consisted of 3 level that namely ethanol 75%, 85% and 95%. The second factor was the time of extraction which consisted of 4 level namely 5, 10, 15, and 20 minutes. The data were analyzed by analysis of variance and continued with Tukey test. The results showed that the concentration and extraction time had a very significant effect on yield, total phenolic, total flavonoid and antioxidant capacity of cocoa pod husk extract. Interactions between treatments had a very significant effect on total phenolic and total flavonoid. Interaction on antioxidant capacity had a significant effect, but did not significantly effect to the yield of cocoa pod husk extract. The best treatment showed that extraction by ethanol 95% for 15 minutes using MAE which the characteristic yield was 11.85±0.84% total phenol was 149.36±0.92 mg GAE/g extract, total flavonoid was 91.03±2.31 mgQE/g extract, antioxidant capacity was 136.69±0.19 mg GAEAC/g extract.
 Keyword : Ethanol concentration, time extraction, antioxidants, cocoa pod husk.

Cacao Pod Husk Extract Phenolic Nanopowder-Impregnated Cellulose Acetate Matrix for Biofouling Control in Membranes.

The ultrafiltration membrane process is widely used for fruit juice clarification, yet the occurring of fouling promotes a decline in process efficiency. To reduce the fouling potential in the membrane application in food processing, the use of natural phenolic compounds extracted from cocoa pod husk is investigated. The cocoa pod husk extract (CPHE) was prepared in phenolic nanoparticles form and added into the polymer solution at varying concentrations of 0.5 wt%, 0.75 wt%, and 1.0 wt%, respectively. The composite membrane was made of a cellulose acetate polymer using DMF (dimethylformamide) and DMAc (dimethylacetamide) solvents. The highest permeability of 2.34 L m−2 h−1 bar−1 was achieved by 1.0 wt% CPHE/CA prepared with the DMAc solvent. CPHE was found to reduce the amount of Escherichia coli attached to the membranes by 90.5% and 70.8% for membranes prepared with DMF and DMAc, respectively. It is concluded that CPHE can be used to control biofouling in the membrane for food applications.

PENGARUH SUHU DAN WAKTU MASERASI TERHADAP AKTIVITAS ANTIOKSIDAN EKSTRAK KULIT BUAH KAKAO (Theobroma cacao L.)

Cocoa pod husk is a by-product of cocoa processing which is quite abundant and has not been used optimally. Cacao pod husk can be used more optimally by extracting, its content of polyphenol compounds which can be used as natural antioxidants. The aim of this study were to determine the effect of temperature and maceration time of cocoa pod husk extract as a source of antioxidants and to determine the best type of temperature and maceration time to produce cocoa pod husk extract as a source of antioxidants. This experiment was designed by using factorial randomized block design. The first factor was type of maceration temperature consisting of 30±2°C, 45±2°C and 60±2°C. The second factor was maceration time, which were done for 24, 36 and 48 h. The data were analyzed with analysis of variance and continued with the Tukey test. The results showed that the temperature and time of maceration had a very significant effect on yield, total phenolic and antioxidant capacity of cocoa pod husk extract. Interactions between treatments had a very significant effect on total phenolic and antioxidant capacity but did not significantly affect the yield of cocoa pod husk extract. The best treatment for producing cocoa pod husk extract as a source of antioxidants was using maceration temperature 60±2°C and maceration time for 36 h with yield characteristics 5,28±0,15%, total phenolic at 168.16±0,06 mg GAE/g and capacity antioxidant 130.94±0.84 mg GAEAC/g.
 Keywords: Cocoa pod husk, extraction, temperature, time, antioxidants.

In vitro studies of antioxidant, antidiabetic, and antibacterial activities of Theobroma cacao, Anonna muricata and Clitoria ternatea

The present study explored the total phenolic contents, antioxidant, antibacterial, and antidiabetic activities of four Indonesian plants, namely Theobroma cacao pod husk (yellow variant and purple variant of pod husk), Annona muricata leaves, and Clitoria ternatea flower. Antioxidant properties of the extracts and fractions were evaluated using a DPPH (2,2-diphenyl-1-picryl-hydrazyl) assay, and their antibacterial activity against Staphylococcus aureus and Escherichia coli by the microdilution method using resazurin as a colouring agent. The antidiabetic assay was performed by an α-glucosidase inhibitory activity using a glucosidase from Saccharomyces cerevisiae. The results revealed that the pod husk extract of T. cacao of both variants (yellow and purple) had good antioxidant activity and antidiabetic activity with IC50 as of 41.3 and 44.5 μg/mL for the DPPH assay, and 41.6 and 27.7 μg/mL for the α-glucosidase inhibitory assay, respectively. Meanwhile, for antibacterial test, T. cacao pod husk of both variants also possessed the most active extracts with MIC (minimum inhibitory concentration) as of 0.62 mg/mL against S. aureus. Moreover, the FTIR (Fourier Transform Infrared Spectroscopy) analysis suggested the presence of phenolic molecules containing in the extracts. This study revealed that T. cacao pod husk could be a potentially rich source of naturally occurring antibacterial, antioxidant, and antidiabetic medicine.

Synthesis of ZnO nanoparticles using Theobroma cacao L. pod husks, and their antibacterial activities against foodborne pathogens

Zinc oxide nanoparticles (ZnONPs) with antimicrobial properties have potential applications in the food industry. In the present work, the biosynthesis of ZnONPs was carried out using Theobroma cacao L. pod husk extract. UV-Vis spectroscopy confirmed NP formation by the presence of a peak in the range of 390 - 360 nm. Different parameters affecting the biosynthesis were assessed. A temperature of 28°C and pH of 7 were the best conditions for the formation of small-sized NPs. The antibacterial activity of ZnONPs was evaluated using the resazurin microtiter assay. The minimum inhibitory concentrations of ZnONPs for Escherichia coli and Staphylococcus aureus were 6.25 and 12.5 μg/mL, respectively. ZnONPs were more potent than chloramphenicol, suggesting that they are effective against chloramphenicol-resistant bacteria. Based on scanning electron microscopy and transmission electron microscopy, the ZnONPs were irregular in shape, with an average size of 81 nm. The results of the present work provide a simple, cost-effective, and eco-friendly method for the large-scale production of ZnONPs. Moreover, the study highlights the potential of ZnONPs as an antimicrobial agent that can be applied to food packaging systems, textiles, and medical devices.

Ekstrak Kulit Buah Kakao (Theobroma cacao L.) Limbah Perkebunan Berpotensi sebagai Akarisida Alami Terhadap Caplak Boophilus microplus

This study was aims to exploit the potential waste of cocoa pod (Theobroma cacao L.) as a natural acaricide in ticks (Boophilus microplus). The study was conducted with a two-variable completely randomized design (CRD) with two repetitions. The sample does not differentiate between sex, body weight, and length of about 0.8 - 1 cm. Each 5 ticks were treated as follows: treat 1 extract of 10% cocoa pod husk; treatment 2 20% cocoa pod skin extract; treatment 3 30% cocoa pod husk extract; treatment 4 negative control; treat 5 positive controls. In the first variable the tick was treated with methanol extract, while in the second variable the ethanol extract was treated with 10%, 20%, 30% respectively, negative control and positive control. In treatment I used 25 ticks plus treatment II 25 ticks. Each treatment was repeated to avoid data bias so that a total of 100 ticks were used. The results showed that the cocoa pod husk extract had the best killing power at a concentration of 30% with methanol solvent at 6.21 hours while ethanol solvent at 5.91 hours. The use of extraction materials has not been able to compensate for the use of synthetic materials that can kill in minutes.