Formulating functional confectionery: biochemical characterization and antioxidant properties of Terminalia arjuna-infused milk chocolate

Introduction

Medicinal plants are the conventional source of medicine for healthcare. According to World Health Organization (WHO) 2002 report, about 80% of the global population relies on traditional medicine for some healthcare purposes and nearly 60% Indian rural population use herbal remedies as their main treatment. The interest in medicinal plants attributed to their effectiveness, no or fewer side effects, cultural acceptance, increasing healthcare costs, etc. Medicinal plants possess compounds that have interacting factors, including therapeutic potential, bioactive compounds, pharmaceutical and as a nutraceutical supplement. Several studies reported the medicinal plants potential and investigation for development of value added or functional products. Terminalia arjuna (Roxb.) Wt. & Arn. is one such tree, belonging to the family Combretaceae. It is a deciduous tree native to the Australia, Asia and Africa (Sivalokanathan et al., 2006) [1] . Whereas in India, it is present along the river banks across Maharashtra, Madhya Pradesh, West Bengal, Uttar Pradesh and Odisha. Terminalia arjuna has been reported to possess biological activities, antiviral, anticancer, antibacterial and antimalarial (Cock, 2015) [2] , wound-healing properties (Amalraj & Gopi, 2017). The different parts of the Terminalia arjuna tree (viz. fruits, leaves, stem bark) exhibit distinct therapeutic profiles. phytochemical density in Terminalia arjuna supports for its development as a plant-derived functional ingredient. The study by Mitra et al. (2005) [3] reported that stem bark of Terminalia arjuna is rich in bioactive compounds, including triterpenoids, glycosides, flavonoids, tannins, and various minerals. Despite its immense therapeutic potential, the direct consumption of Terminalia arjuna bark is limited because of its bitter taste, coarse texture and poor consumer acceptability. Chocolate is the widely consumed confectionery product across globe. It is a complex matrix of cocoa solids, sugar and milk components dispersed in continuous fat phase of cocoa butter and milk fat. Depending on its composition they are generals classified as dark, milk and white chocolate. Chocolate possess the favorable sensory properties as well as market acceptance, it offers a suitable platform for incorporating bioactive compounds, enabling the formation of nutritionally enhanced, value-added products (Singh et al., 2024) [4] . The incorporation of herbal plant material like Terminalia arjuna is therefore and emerging approach for value-added functional product. In this research, the milk compound chocolate was used as a base ingredient and the Terminalia arjuna bark powder was incorporated by substituting it's part.

Materials and Methodology

Materials

The research study was conducted using raw materials such as Terminalia arjuna bark was collected from campus of VNMKV, Parbhani. The required milk chocolate was procured from the local market of Parbhani. Food grade ethanol was made available in Department of Food Chemistry and Nutrition, College of Food Technology, VNMKV, Parbhani.

Methodology

A) Cutting, cleaning, size reduction, and sieve analysis 1. Cutting: The stem bark of Terminalia arjuna was detached from the matured tree with sterile knife. The collected sample was authenticated by appropriate authority before experimental processing.

Cleaning:

The collected bark was washed thoroughly to remove the adhering dust, soil and extraneous matter. Then was oven dried at controlled temperature.

Pulverization:

The pulverization of the dried bark was carried out using the laboratory-scale grinder.

Sieving:

The grounded Terminalia arjuna bark powder was passed through a specific mesh size to gain homogeneous, and uniform particle size.

B) Pre-treatment (alcohol treatment)

The 150 ml of 70% Ethanol (food grade) solution was prepared and placed in extraction vessel and the Terminalia arjuna bark powder (30 g) was added. Cold maceration was carried out at room temperature (25 ± 2 °C) for 24 hr. with intermittent shaking. The mixture was filtered and the resulting filtrate was concentrated and dried in hot air oven at 40-45 °C. The dried extract was ground into a homogeneous powder as mentioned by (Parmar et al., 2017) [5] .

C) Preparation of functional chocolate

Sensory evaluation of functional milk chocolate

The sensory characteristics was studied by the 10 trained panelists using the 9-point hedonic scale for prepared samples (T 0 , T 1 , T 2 , T 3 ).

The sensory attributes evaluated included color, appearance, taste, flavor and overall acceptability.

Evaluation of chemical composition of functional chocolates

The moisture content by hot air oven method (AOAC, 2000) [6] , fat using soxhlet apparatus (AACC, 2000) [7] , protein estimated by Micro-kjeldhal apparatus as described in AACC (2000) [7] , ash, crude fiber AOAC (2000) [6] and carbohydrate by difference method was evaluated.

Evaluation of functional properties of functional chocolates

The Total Phenolic Content (TPC) was determined using Folin-Ciocalteu colorimetric method as described by Sharma et al. (2023). Total Flavonoid Content (TFC) aluminium chloride method outlined by Chang et al. (2002) [8] . Tannin was determined using tritrimetric protocol given by Sharma et al. (2023). Saponin determined gravimetrically using the protocol given by Obadoni and Ochuko (2001). For alkaloid and antioxidant activity, gravimetric method described by Harborne (1998) [9] and DPPH scavenging activity in percent using method by Brand Williams et al., 1995 [10] .

Statistical analysis

The results were obtained in triplicates (n = 3) and expressed as a mean ± standard deviation.

Result and Discussion

Organoleptic evaluation of functional milk chocolate

The organoleptic evaluation of functional chocolate sample were evaluated for control and other prepared samples. The all samples were evaluated for color and appearance, taste, flavor, texture and overall acceptability. The sample T 2 out scored all the other samples, securing the highest mean scores for color and appearance (8.8), taste (8.3), flavor (8.1), texture (8.3) and overall acceptability (8.4). The control sample (T 0 ) and selected sample (T 2 ) were further evaluated for nutritional and functional analysis. The proximate composition of prepared functional chocolate was carried out to determine the changes occurred by addition of Terminalia arjuna bark powder in chocolate. The control sample (T 0 ) and selected sample (T 2 ) were evaluated. The chemical composition date revealed that, the moisture content rises from 0.55% in T 0 to 0.62% in T 2 sample, as the Terminalia arjuna bark powder is hygroscopic in nature and in traps the atmospheric air during mixing and processing stages. The fat content was found to be at 31.43% in T 0 and 30.51% in T 2 . The reason is that as the bark powder is incorporated in chocolate, a part of it chocolate was replaced. The arjuna powder as very little amount of fat, by substituting the fat rich matrix of chocolate it lowers overall fat percentage in finished chocolate. The similar trend was observed in case of protein at 7.60% in T 0 and 7.40% in T 2 and for fiber T 0 at 1.70% and 1.60% for T 2 sample. The ash content increased due to the fact that the Terminalia arjuna bark powder is rich in minerals like calcium and magnesium, that directly increases the ash content of the functional chocolate. The carbohydrate content increased from 57.47% in T 0 to 58.01% in T 2 , due to the reason that bark of Terminalia arjuna has high carbohydrate fraction. The functional properties of the standardized functional chocolate are tabulated in Table 4. The Total Phenolic Content of T 0 sample was found to be 2.90 mg GAE/g and T 2 sample was found to be 5.28 mg GAE/g. While the Total Flavonoid Content was 6.37 mg QE/mg for T 0 sample and 11.24 mg QE/mg for T 2 sample. The increase in the values of TPC and TFC indicated the evidence of polyphenols in Terminalia arjuna bark powder was migrated successfully into the functional chocolate. The tannin content was found at 0.47% in T 0 to 1.31% in T 2 . While the addition of Terminalia arjuna bark powder introduced measurable trace level of saponins 0.030% in the final chocolates an active cardiotonic compound that was completely absent in control samples. This targeted addition of tannins and saponins successfully drove up the overall anti-oxidant activity, which climbed to 24.60% in T 2 from 23.15% in T 0 . The antioxidant activity of compound chocolate was less compared to cocoa due to the processing steps, while the cocoa itself is rich in antioxidants. In contrast, the alkaloid content was 0.050% in T 0 and 0.052% in T 2 , as the alkaloids content in Terminalia arjuna bark powder is negligible and the cocoa solids drive the baseline of the chocolate. These balanced functional updates line up well with established herbal food delivery frameworks published by Ghotkar et al.

Proximate of prepared functional chocolate

Determination of functional properties of standardized chocolates

Mineral composition of standardized chocolates

Conclusion

The research is going and showed immense potential of the medicinal plants for functional food applications. The Terminalia arjuna is rich source of essential nutrients as well as bioactive compounds, incorporating it in food make it an excellent ingredient for enriching food with it's beneficial nutrients. The functional chocolate was prepared by giving alcohol treatment to Terminalia arjuna bark powder. The chocolate with 2% addition of Terminalia arjuna bark powder showed enhancement in nutrients, mineral and functional profiles of it. The confectionery products are willingly consumed by children, teenagers and adult. Therefore, the standardized functional chocolate offers a highly viable strategy to enrich them with antioxidant, cardioprotective and nutrient-dense properties.

References

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