Optimization of the Production Process of Bishop’s Crown Pepper (Capsicum baccatum var.) Hydroalcoholic Extract

This study developed and evaluated the efficiency of methods to produce hydroalcoholic extract of Bishop’s Crown pepper. Analyses were conducted to investigate the effects of independent variables (ethanol concentration of the extracting solution and resting time) over the dependent variables (percent yield, acidity, carotenoids, phenolic compounds, anthocyanin content and vitamin C). The interactions between the variables were studied through seven factorial experiments. Percent yield (Y%), carotenoids, phenolic compounds and vitamin C were the most satisfactory responses found in this research. The ethanol concentration of the extracting solution (ES) presented significant effect in the Y%, showing better results for extracts produced with a 55%-ethanol ES, while carotenoids, phenolic compounds and vitamin C displayed significant differences according to the ethanol concentration, being these substances found in higher concentrations in extracts produced with the use of an 85%-ethanol ES. Therefore, the higher the ES ethanol concentration, higher is the retention of the bioactive compounds.


Optimization of the Production Process of Bishop's Crown Pepper
(Capsicum baccatum var.) Hydroalcoholic Extract

Introduction
There is a large variety of species of peppers of the genus Capsicum, which belongs to the Solanaceae family. Their origin is connected to the tropical, and especially the humid, zones in America. Peppers have nutritional and pharmacological properties, and are used in alternative medicine (Kim et al., 2010;Lahbib et al., 2017;Meghvansi et al., 2010).
The Bishop's Crown pepper (Capsicum baccatum var.), also known as Cambuci pepper, is one of the most consumed condiments in South America, however there are few studies and data in the literature regarding its chemical composition and possible biological properties. Recent studies have shown that extracts produced from C. Baccatum have antioxidant properties (Allemand et al., 2016;Zimmer et al., 2012).
Nowadays, there is relevant interest in producing and studying extracts from natural sources that have bioactive properties and potential to be used in the food industry, for instance, in the replacing synthetic food colorings for natural ones, and also for their potential antioxidant agents. The final quality of the extracts is related to all the factors to which the material is exposed during the production process, making it necessary to study each factor and their impact on the final product individually.
The use of a planning matrix and/or factorial experiments is of great interest in processes of various natures, once experiments are conducted and tested for all possible combination, being analyzed the levels of each factor and variable, as well as identifying each level as an experiment itself. Due to that, factorial experiments aim to define in a quantitative manner the impact of each factor on the process, making it possible for researchers to reach results with a lower number of experiments, leading to fewer expenses, time investment, and resources (Neto et al., 2010).
During the extraction of bioactive compounds from vegetable products several factors impact the final-product characteristics, such as the shape and size of the material, the type and concentration of the extracting solution (ES) used, as well as the time during which the sample and the ES remain in contact. Therefore, the extracts obtained may contain different concentrations of bioactive compounds (Vongsak et al., 2013).
Facing the challenges exposed, this study developed and evaluated the efficiency of methods to produce hydroalcoholic extract of Bishop's Crown pepper, through the investigation of the effects of the ethanol concentration of extracting solutions (ES) and resting time, over percent yield, acidity and bioactive compounds (carotenoids, phenolic compounds, anthocyanin content and vitamin C).

Raw Material and Research Site
The Bishop's Crown peppers were acquired in the local commerce of the city of Pombal, Paraíba, Brazil, being then transported to the Food Chemistry and Biochemistry Laboratory of the Federal University of Campina Grande campus in the same city where samples were acquired.

Production of Bishop's Crown Pepper Hydroalcoholic Extract
From the peppers acquired, only the ones in perfect maturation stage and with no visible physical damage were selected to go under a cleaning process for removing any residues and potential microorganisms in the fruits. For the production of the extract, the peppers had seeds removed and were processed in a mixer (M-08) to obtain a homogeneous paste, which was added to the extracting solution (ethanol) in different concentrations (55%, 70%, and 85%) and resting times (8 h, 16 h, and 24 h), as detailed in the Table 1.

Calculating the Percent Yield of the Extracts
The percent yield (Y%) was calculated according to Rodrigues et al. (2011), considering the relation between the volume of the final extract (VFE) and the volume of the initial extract (VIE) before filtration and evaporation of the solvent. The following equation was used:

Chemical Characterizing of the Extracts
The extracts were characterized in triplicate for the following physicochemical coatings: total titratable acidity and vitamin C according to the methodology of the Adolfo Lutz Institute (Brazil, 2008). Carotenoids were determined according to Lichtenthaler (1987), the anthocyanins were determined according to the Francis method (1982) and the determination of total phenolic compounds was performed by the Folin-Ciocalteu method, described by Waterhouse (2006).

Statistical Analysis
The studies of the dependent variables were treated used using the graphs of parentheses and the contour surface methodology. ANOVA was applied to test the meaning of the interaction process (p < 0.05). To visualize the