Antibacterial Activity of the Cream Preparation from Theobroma cacao L. Pod Aqueous Extract

Abstract

Background and Objectives. While Theobroma cacao L has long been utilized in the food, cosmetic, and pharmaceutical industries, it was also found to possess antibacterial activity. The beans comprise 10% of the fruit, while the remaining 90%, consisting of pods, is considered waste. It was reported that the pods possess antibacterial activity, and if utilized for this purpose, T. cacao pods will no longer be considered as waste. The aim of this study was to evaluate the antibacterial activity of the cream formulated from the aqueous extract of T. cacao L pods.
 Methods. The milled T. cacao pods were extracted using distilled water at 4°C for 24 hours. The crude extract was subjected to liquid-liquid partitioning using hexane, ethyl acetate, and n-butanol. Phytochemical screening was performed to identify the constituents present in the extract and its fractions. The extract and its fractions were tested against Staphylococcus aureus, Staphylococcus epidermidis, and Pseudomonas aeruginosa. Determination of IC50 using 3,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) Reduction Assay was used to evaluate the antibacterial activity. The extract with the highest yield and the highest antibacterial activity were formulated into a cream. T. cacao cream was evaluated with quality control tests for creams and emulsions. Acute skin irritation test was performed on the T. cacao cream to assess skin irritability upon application on adult male albino rabbits.
 Results. T. cacao crude extract and its fractions possessed antibacterial activity. Among the fractions tested, n-butanol fraction had the highest activity against S. aureus, S. epidermidis, and P. aeruginosa. There was a significant difference between the fractions tested on the three bacterial strains (p<0.05). Although n-butanol fraction had the highest activity, the actual yield obtained after extraction was 0.95%. Since T. cacao aqueous extract also exhibited good antibacterial activity, it was chosen for the formulation study. There was no significant difference between the IC50 of the T. cacao crude extract and the IC50 of T. cacao cream, hence formulating it into a cream did not affect the antibacterial activity of the extract.
 Conclusion. T. cacao pod extract, as well as its fractions, possessed antibacterial activity against three bacterial strains. The T. cacao cream produced was a water-in-oil, non-irritant cream with antibacterial activity, and with acceptable physical attributes.