Physicochemical Properties and Antioxidant Activity of Powdered Beverage Based on Andaliman (Zanthoxylum acanthopodium) and Kuweni (Mangifera odorata)

Abstract

Integrating andaliman (Zanthoxylum acanthopodium) and kuweni (Mangifera odorata) into a functional powdered beverage provides a novel method for maximizing the bioactive potential of these indigenous ingredients. This study aimed to evaluate the physicochemical properties, antioxidant activity, and safety of powdered beverages formulated using varying proportions of andaliman and kuweni: F1 (25% andaliman:75% kuweni), F2 (50% andaliman:50% kuweni), and F3 (75% andaliman:25% kuweni). The results showed that the solubility (95.04% in F1 to 97.74% in F3) and pH (3.74 in F1 to 4.21 in F3) increased with higher andaliman proportions. The total flavonoid content ranged from 21.18 mg/g (F1) to 25.06 mg/g (F3), whereas the antioxidant activity (expressed as IC₅₀) ranged from 114.14 ppm (F1) to 88.20 ppm (F3), demonstrating a strong antioxidant potential. All formulations met the Indonesian National Standard for traditional powdered beverages, ensuring microbial safety and the absence of heavy metals. Our findings highlight the potential of andaliman and kuweni as key ingredients in the innovation of functional beverages, contributing to health-promoting products and the valorization of North Sumatra’s food heritage.

INTRODUCTION

Functional foods, which provide health benefits beyond basic nutrition, have attracted increasing interest because of their potential to prevent chronic illnesses and promote overall health and well-being. This growing demand has driven innovation in food products using functional ingredients, particularly those derived from natural and indigenous sources (Cong et al., 2020; Cuamatzin-García et al., 2022). Among these ingredients, using bioactive-rich plant components in developing beverages has shown promising potential in meeting modern consumer preferences for health-promoting and culturally significant products (Orrù et al., 2018).

Andaliman (Zanthoxylum acanthopodium) and kuweni (Mangifera odorata), which are two indigenous ingredients from North Sumatra, Indonesia, exhibit notable functional properties. Andaliman is widely known for its distinctive citrus-pepper flavor along with high phenolic and flavonoid contents, which contribute to its potent antioxidant activity (Adrian et al., 2023). It is traditionally used in culinary and medicinal contexts, reflecting the local reliance on natural resources for health and flavor (Santoso et al., 2023). Kuweni, a variety of mango, contains high amounts of vitamins A and C, polyphenols, and carotenoids, which contribute to its antioxidant capacity and the sweet and sour taste profile (Lasano et al., 2021; Lestari et al., 2021).

Integrating these two ingredients into a functional powdered beverage offers a novel approach to functional food development. Moreover, the distinct flavor of andaliman complements the sweetness and acidity of kuweni, thereby creating a refreshing and complex beverage with sensory and nutritional appeal (Natasutedja et al., 2020). Previous studies have shown that fruit blends containing mango can improve the nutritional value and consumer acceptance, making kuweni an ideal candidate for the formulation of beverages (Liguori et al., 2020; Arum and Ani, 2021).

However, despite their individual benefits, few studies have explored the synergistic potential of andaliman and kuweni, particularly in terms of antioxidant activity, solubility, pH stability, and overall functionality in the form of a beverage. Moreover, there are challenges with regard to the formulation process, including balancing the flavor profiles and ensuring that microbial and heavy metal safety standards are met (Zhou and Liu, 2024). Addressing these challenges presents an opportunity to develop an innovative functional powdered beverage that valorizes local ingredients while meeting the consumer demand for healthy and natural products.

Therefore, the present study aimed to evaluate the physicochemical properties and antioxidant activity of a powdered beverage developed using andaliman and kuweni, including assessments of heavy metal and microbial contaminants to ensure its safety and functionality. This study presents a novel approach by formulating a powdered beverage that leverages the unique bioactive profiles of andaliman and kuweni.

MATERIALS AND METHODS

Materials

Andaliman, kuweni, sugar, and egg white were purchased from the Medan Metropolitan Trade Centre Wholesale Market, Medan, Indonesia. Maltodextrin (Zhucheng Dongxiao Biotechnology Co., Ltd.) was obtained from Makmur Sentosa Food Chemical, Tangerang, Indonesia. All chemicals used in this study were of analytical grade (Merck). The microbiological media included plate count agar (PCA), lauryl sulfate tryptose (LST) broth, and Brilliant Green Lactose Bile Broth (BGLB 2%) (Oxoid Ltd.). All solutions were prepared using distilled water, and all glassware and equipment were sterilized prior to use.

Sample preparation

Andaliman powder was prepared by cleaning 50 g of berries. Afterward, they were extracted twice using 100 mL of 96% ethanol. The extract was initially filtered through a 100-mesh sieve and then with Whatman No. 41 filter paper using a vacuum pump (Joanlab Equipment Co.) to ensure clarity. Prior to drying, 15% maltodextrin was dissolved in water equivalent to 50% of the extract’s volume and thoroughly mixed with the extract. Subsequently, the mixture was dehydrated using a dehydrator (CV Wiratech Jaya Mandiri) at 50°C for 12 h. The dried flakes were then ground into powder, obtaining a yield of 56.23%. Meanwhile, kuweni powders were prepared by blending 100 g of fresh fruit flesh. The fresh fruit was peeled, deseeded, and blended into a smooth puree, which was then strained through a 100 mesh sieve. The filtrate was mixed with 10% egg white and 15% maltodextrin. Thereafter, the mixture was processed into a foam and dried under the same conditions as the andaliman powder. The dried product was ground into powder, obtaining a yield of 26.9%. These powders were combined into formulations with varying andaliman-to-kuweni-powder ratios. The andaliman-to-kuweni powder ratios were 25:75 (F1), 50:50 (F2), and 75:25 (F3). The total weight of each formula was 12 g, including 8 g of powdered sugar to standardize the formulations. Each formulation was analyzed in duplicate for all tests, resulting in six experimental units for physicochemical, antioxidant activity, microbial, and heavy metal analyses. This duplication ensured the accuracy and reproducibility of data.

Analysis of physicochemical properties

Solubility measurement: The solubility measurement was performed in accordance with the method of Adhayanti and Ahmad (2020) with slight modifications. A 5 g sample of the powdered beverage from each formulation (F1, F2, and F3) was weighed as the initial weight. Subsequently, the sample was dissolved in 200 mL of water at 95°C and stirred 15 times before being filtered through a preweighed filter paper. The filter paper was oven-dried at 105°C for 3 h, cooled in a desiccator, and weighed until a constant weight was achieved. The solubility (%) was calculated using the following equation:

$$\text{Solubility (\%)=}\frac{{\text{W}}_{\text{initial}}\text{-}\left({\text{W}}_{\text{residue+filter}}{\text{-W}}_{\text{filter}}\right)}{{\text{W}}_{\text{initial}}}\times 100$$ (Eq. 1)

where W_initial is the initial weight of the powdered sample (g), W_{residue+filter} is the weight of the dry residue along with filter paper (g), W_filter is the weight of the empty filter paper (g).

pH measurement: The pH analysis was performed in accordance with the method of Trimedona et al. (2022). A pH meter was turned on and allowed to stabilize until it displayed accurate readings. Moreover, it was calibrated using a buffer solution and distilled water. The pH measurement was performed by immersing the pH meter electrode in the solution until a stable pH reading was obtained. The sample was mostly soluble. To ensure accurate pH measurement, the solution was allowed to settle, and a clear supernatant was used for measurement to prevent interference from sediments.

Bulk density measurement: The bulk density was measured in accordance with the method of Romulo and Aurellia (2024). The sample was carefully poured into a measuring cylinder and compacted until its volume reached 100 mL. Then, the entire sample was removed from the cylinder and weighed. The bulk density of the material (%) was expressed in grams per milliliter×100.

Hygroscopicity measurement: The hygroscopicity was measured in accordance with the method of Li et al. (2021). Approximately 1 g of the sample was weighed and placed in a Petri dish at room temperature. The Petri dish was stored with a saturated sodium chloride solution to maintain a relative humidity of 75% for one week. The hygroscopicity (%) was calculated using the following formula:

$$\text{Hygroscopicity (\%)=}\frac{\text{WI\%+MC\%}}{\text{100+WI\%}}\times 100$$ (Eq. 2)

where MC% is the initial moisture content of the sample and WI% is the water intake percentage. The WI% value was calculated using the formula:

$$\text{WI\%=}\frac{\text{Weight of sample after equilibrium}-\text{weight of sample}}{\text{Weight of sample}}\times 100$$ (Eq. 3)

Moisture content measurement: The moisture content was measured in accordance with the Indonesian National Standard (SNI) for traditional powdered beverages (BSN, 1996). About 1 to 2 g of the sample was placed in a preweighed, lidded weighing bottle. Then, the sample was dried in an oven at 105°C for 3 h to remove moisture. After drying, the sample was cooled in a desiccator to prevent moisture absorption from the surrounding air. Once cooled, the sample was weighed again. This process was repeated until a constant weight was obtained, ensuring that all moisture has been removed. Subsequently, the moisture content was calculated based on the weight loss after drying:

$$\text{Moisture content \%=}\frac{{\text{W}}_{\text{1}}{\text{-W}}_{\text{2}}}{{\text{W}}_{\text{1}}}\text{×100}$$ (Eq. 4)

where W₁ is the initial weight of the sample before drying, and W₂ is the weight after drying.

Ash content measurement: The ash content was measured using the dry ashing method as described in the SNI (BSN, 1996). About 2 to 3 g of the sample was placed in a porcelain crucible with a known weight. Then, the crucible was placed over a burner flame before being ashed in an electric furnace at a maximum temperature of 550°C until complete ashing was achieved. Occasionally, the furnace door was slightly opened to allow oxygen to enter and facilitate combustion. Once the ashing process was completed, the crucible was cooled in a desiccator and weighed until a constant weight was obtained.

$$\text{Ash content (\%)=}\frac{{\text{W}}_{\text{ash}}}{{\text{W}}_{\text{initial}}}\times 100$$ (Eq. 5)

where W_ash is the weight of the residue after ashing (i.e., ash weight), and W_initial is the weight of the sample before ashing.

Total sugar measurement: The total sugar was measured in accordance with the SNI standard method (BSN, 1996). About 5 g of the sample was accurately weighed and placed into a 500-mL Erlenmeyer flask. Then, 200 mL of 3% HCl solution was added, and the mixture was boiled for 3 h using a reflux condenser. After cooling, the solution was neutralized with 30% NaOH solution, and a small amount of 3% CH₃COOH was added to slightly acidify the solution. Then, the solution was transferred into a 500-mL volumetric flask and diluted to the mark with distilled water before being filtered. Next, 10 mL of the filtrate was pipetted into a 500-mL Erlenmeyer flask, to which 25 mL of Luff-Schoorl reagent and a few boiling chips, along with 15 mL of distilled water, were added. The mixture was heated with a constant flame until boiling was achieved within 3 min (using a stopwatch). Boiling was maintained precisely for 10 min (measured from the onset of boiling). Thereafter, the solution was rapidly cooled in an ice bath. Once cooled, 15 mL of 20% KI solution and 25 mL of 25% H₂SO₄ solution were added slowly. Subsequently, the solution was titrated immediately using 0.1 N Na₂S₂O₃, with 0.5% starch solution as an indicator. A blank titration was also performed for reference. The total sugar was calculated using the formula:

$$\text{Total sugar (\%)=}\frac{{\text{W}}_{\text{1}}\text{×fp}}{\text{W}}\times 100$$ (Eq. 6)

where W₁ is the glucose equivalent equivalent (mg) obtained from the Luff-Schoorl table based on the volume (mL) of sodium thiosulfate (Na₂S₂O₃) used during titration, W is the weight of the sample (mg), and fp is the dilution factor.

Measurement of total flavonoid content

The total flavonoid content was measured in accordance with the method of Silalahi et al. (2019). About 0.5 mL of the sample extract was mixed with 0.3 mL of 5 g/100 mL NaNO₂ solution and incubated at room temperature for 5 min. Thereafter, 0.3 mL of 10 g/100 mL AlCl₃ solution was added. After 1 min, the mixture was neutralized with 2 mL of 1 M NaOH. The absorbance of the solution was measured at 510 nm using an ultraviolet-visible (UV-Vis) spectrophotometer (Shimadzu). Quercetin was used as the standard, and different concentrations (0－100 µg/mL) were prepared for calibration, achieving an R² value of 0.9993. The results were expressed as mg quercetin equivalents per 100 g of the sample, reported as the mean of duplicate measurements.

Measurement of antioxidant activity

The antioxidant activity was measured in accordance with the method of Lestari et al. (2021) with modifications. A 2,2-diphenyl-1-picrylhydrazyl (DPPH) stock solution was prepared by dissolving 10 mg of DPPH powder in 100 mL of hot water and stirring until homogeneous. For the blank solution, 10 mL of the DPPH stock solution was pipetted into a 25-mL volumetric flask, diluted with hot water to the mark, and homogenized to achieve a final concentration of 40 ppm. The absorbance of the blank solution was measured using a UV-Vis spectrophotometer (Shimadzu) at wavelengths ranging from 400 nm to 600 nm, with ethanol p.a. serving as the blank. About 0.1 g of each sample was weighed using an analytical balance and dissolved in distilled water in a volumetric flask. To prepare the stock solution, hot distilled water (100 mL) was added to the flask. Next, 25-, 50-, 75-, 100-, and 125-µL aliquots from the stock solution were pipetted to prepare concentrations of 5, 10, 15, 20, and 25 ppm, respectively. Each aliquot was mixed with 2 mL of 40-ppm DPPH solution and diluted with distilled water to a total volume of 5 mL. Then, the samples were vortexed, incubated at 37°C for 30 min, homogenized, and left for 30 min before measuring the absorbance at the maximum wavelength of 517 nm using a UV-Vis spectrophotometer (Shimadzu).

The antioxidant activity was expressed as the percentage of DPPH radical inhibition, which was calculated using the following formula:

$$\text{\% Inhibition=}\frac{\text{Blank absorbance}-\text{Sample absorbance}}{\text{Blank absorbance}}\times 100$$ (Eq. 7)

The IC₅₀ value, which represents the sample extract concentration needed to reduce DPPH radicals by 50%, was determined from the linear regression equation between the percent inhibition and the sample concentrations. The IC₅₀ value was calculated using the following formula: Y=ax+b, where a is the concentration constant, b is the constant, x is the concentration, and Y is the percentage of attenuation. A lower IC₅₀ value indicates higher antioxidant activity.

Heavy metal and microbial contaminant analysis

Contaminant analysis included testing for heavy metals [e.g., lead (Pb), copper (Cu), zinc (Zn), tin (Sn), and arsenic (As)] and microbial contamination [total plate count (TPC) and coliform]. All analytical methods were performed in accordance with the methods described in the SNI for traditional powdered beverages (BSN, 1996).

Pb, Cu, Zn, and Sn content analysis: About 1 to 5 g of the sample was placed into a 250-mL Erlenmeyer flask and then added with 25 mL of HCl solution. Subsequently, the mixture was heated to boiling and maintained at this condition for 5 min. After heating, the solution was cooled and quantitatively transferred into a 50-mL volumetric flask. The solution was diluted to the mark with distilled water, mixed thoroughly, and filtered through a Whatman No. 1 filter paper. A blank solution was prepared using the same reagents but without the sample. Then, the absorbance of the standard series, blank, and sample solutions was measured. A calibration curve was constructed by plotting the absorbance (Y-axis) against the concentration in ppm (X-axis). This calibration curve was used to calculate the metal content in the sample. The metal content in the sample (expressed in mg/g) was calculated using the following formula:

$$\text{Metal content (mg/g)=}\frac{\text{μg metal/mL from calibration curve×V}}{\text{m}}\times 1,000$$ (Eq. 8)

where V is the volume of the solution (mL) and m is the mass of the sample (g).

Arsenic content analysis: A 5 g sample was weighed into a digestion flask and mixed with 25 mL of concentrated H₂SO₄ 18N, 20 mL of HNO₃ 7N, 1 mL of 2% Na₂MoO₄ solution, and 3 to 6 boiling stones. The flask was connected to a reflux condenser and heated for 1 h. Heating was stopped for 15 min, and then 20 mL of a mixed acid solution (HNO₃-HClO₄, 1:1) was added through the condenser. The solution was further heated at a high temperature until white fumes appeared and was then maintained for 10 min before cooling. Next, 10 mL of water was carefully added through the condenser while shaking the flask gently. Moreover, heating was resumed for another 10 min. The digestion apparatus was removed, and the condenser was rinsed thrice with 15 mL of distilled water and allowed to cool.

The digested solution was quantitatively transferred to a 100-mL volumetric flask and diluted to the mark with distilled water. A blank solution was prepared using the same reagents without the sample. A standard series was also prepared. About 20 mL of the reducing agent was added to each solution (standard, sample, and blank). The absorbance of each solution was measured using an atomic absorption spectrophotometer without a flame at a wavelength of 253.7 nm. A calibration curve was constructed with the absorbance (Y-axis) versus the concentration (X-axis, in ppm). The As content in the sample (expressed in µg/g) was calculated based on this calibration curve using the following formula:

$$\text{Arsenic content (mg/g)=}\frac{\text{μg arsenic/mL from calibration curve×V}}{\text{m}}\times 1,000$$ (Eq. 9)

where V is the volume of the solution (mL) and m is the mass of the sample (g).

TPC analysis: Sample preparation and homogenization were performed in accordance with standard procedures. Then, 1 mL of each dilution was pipetted into sterile Petri dishes in single (simplex) and duplicate (duplex). About 12 to 15 mL of PCA that had been melted and cooled to 45°C±1°C was poured into each Petri dish within 15 min of the initial dilution. The Petri dishes were gently swirled and rotated in multiple directions (forward, backward, left, and right) to ensure uniform mixing of the sample and medium. A blank control was prepared by mixing dilution water with PCA in the same manner for each sample batch.

The mixture in the Petri dishes was allowed to solidify at room temperature. Thereafter, all plates were inverted and incubated in an incubator at 35°C±1°C for 24 to 48 h. After incubation, the colonies were counted on plates that contained 25 to 250 colonies. The TPC was calculated by multiplying the average colony count per plate by the dilution factor. The results were expressed as the number of colony-forming units (CFU) per gram or per mL of sample.

Coliform analysis: For the presumptive test, the sample was first homogenized in accordance with standard procedures. Then, 1 mL of a 10⁻¹ dilution was pipetted into each of the three test tubes containing 5 mL of LST broth or BGLB with an inverted Durham tube inside. The same procedure was repeated for further dilutions of 10⁻², 10⁻³, and 10⁻⁴, and new sterile pipettes were used for each dilution series. All tubes were incubated at 36°C±1°C for 24 and 48 h. After 24 h, the number of tubes showing gas formation (which indicates the presence of coliform) was recorded, whereas tubes without gas formation were reincubated for an additional 24 h before the final gas observation.

In the confirmed test, one loopful of gas-positive cultures from the LST medium was transferred to tubes containing 10 mL of BGLB 2%. These tubes were also incubated at 36°C±1°C for 24 to 48 h. The presence of gas in the BGLB tubes confirmed the presence of coliform bacteria in the sample. The number of gas-positive tubes from the confirmed test was recorded, and the most probable number (MPN) of coliforms in the sample was determined using the appropriate MPN table.

Statistical analysis

All variables are expressed as the mean±standard error. Statistical analysis was performed to determine significant differences among formulations, with emphasis on the physicochemical properties and antioxidant activity. The results were analyzed using analysis of variance followed by Duncan’s post hoc tests to identify which formulations differed significantly at α=0.05.

RESULTS AND DISCUSSION

Physicochemical properties of andaliman- and kuweni-based powdered beverages

The physicochemical properties of andaliman- and kuweni-based powdered beverages are summarized in Table 1. The findings regarding the solubility, bulk density, pH, hygroscopicity, moisture content, ash content, and total sugar content of the powdered beverage formulations highlight the intricate relationship between the ingredient composition and the physicochemical properties of the final product. The increasing solubility observed with increasing andaliman proportions is particularly noteworthy as it underscores the role of natural sugars in enhancing solute-water interactions. This phenomenon is supported by research indicating that sugars can disrupt the hydrogen bond network of water, thereby facilitating the solubility of hydrophilic compounds (de Souza et al., 2020; Tanaka et al., 2020). The F3 formulation, which contains 75% andaliman, achieved a solubility of 97.74%, which was significantly higher than that of F1 and F2, which had lower proportions of andaliman. This trend not only emphasizes the importance of ingredient selection in beverage formulation but also suggests that the sugar content can be optimized to improve dissolution rates, which is critical for instant beverage applications (Chasquibol et al., 2022).

Table 1.

Physicochemical characteristics of andaliman- and kuweni-based powdered beverages

Characteristic	F1	F2	F3
Solubility (%)	95.04±0.24a	96.20±0.36a	97.74±0.11b
Bulk density (%)	89.09±2.26a	88.20±2.56a	86.51±2.31a
pH	3.74±0.01a	3.95±0.05b	4.21±0.01c
Hygroscopicity (%)	8.25±0.25a	6.94±0.21a	7.32±0.23a
Moisture content (%)	2.08±0.19a	2.08±0.16a	2.50±0.14a
Ash content (%)	0.42±0.06a	0.28±0.01a	0.31±0.03a
Total sugar (%)	69.78±0.01a	67.04±1.01a	66.52±3.13a

Values are presented as mean±standard error from triplicate analysis (n=3).

Different letters in a row indicate significant differences (P<0.05).

F1, 25% andaliman:75% kuweni; F2, 50% andaliman:50% kuweni; F3, 75% andaliman:25% kuweni.

By contrast, the F1 formulation, which contains a higher proportion of kuweni, exhibited reduced solubility because of the presence of hydrophobic compounds that hinder water interactions. This result aligns with findings that the physical properties of ingredients (e.g., the particle size and shape) can significantly influence the solubility and overall quality of the beverage (de Oliveira et al., 2021). The consistent bulk density across formulations, ranging from 89.09% in F1 to 86.51% in F3, indicates that the mixing ratios of andaliman and kuweni do not significantly alter the powders’ packing efficiency. This consistency is advantageous for industrial-scale production to ensure uniformity in packaging and handling, which is crucial for maintaining product stability (Harwood and Drake, 2021).

The pH values of the formulations, which ranged from 3.74 in F1 to 4.21 in F3, indicate that andaliman has lower acidity compared with kuweni. This shift in pH not only affects the sensory profile of the beverages but also suggests that formulations with higher andaliman content may be more palatable to consumers who prefer milder flavors (Premkumar et al., 2022). The sensory attributes of beverages are important for consumer acceptance, and the pH level can significantly influence taste perception. Therefore, the proportions of ingredients need to be carefully considered to optimize flavor profiles in powdered beverage formulations targeted at diverse consumer preferences (Cardinali et al., 2021).

The hygroscopicity, which ranged from 6.94% in F2 to 8.25% in F1, showed no significant differences among formulations, suggesting that andaliman and kuweni have comparable carbohydrate content. A high hygroscopicity can lead to clumping or caking during storage, necessitating careful packaging to maintain product stability. These findings are essential for developing effective packaging strategies that ensure the product’s longevity and quality (Parra-Gallardo et al., 2023).

The formulations’ moisture content, which remained below the SNI limit of 3.0% (BSN, 1996), further enhanced the shelf stability and reduced the risk of microbial growth and spoilage. The slight increase in the moisture content in F3 can be attributed to the inherent capacity of andaliman to retain more water within the formulation (Rezaei and VanderGheynst, 2010). This aspect is crucial for ensuring that the formulations remain suitable for long-term storage while complying with regulatory standards.

The ash content, which ranged from 0.29% in F2 to 0.42% in F1, reflects the mineral composition of the ingredients used. This consistency in moisture and ash contents underscores the stability and nutritional adequacy of the formulations, ensuring compliance with regulatory standards (Ryan et al., 2020). In addition, the total sugar content across formulations showed no significant differences, with values ranging from 69.78% in F1 to 66.52% in F3, which are all well below the SNI maximum limit of 85.0% (BSN, 1996). This controlled sugar level is crucial for catering to health-conscious consumers while maintaining overall product acceptability (de Souza et al., 2020; Velázquez et al., 2021).

These findings collectively demonstrate that the ingredient composition significantly affects the key physicochemical properties and microbial safety of powdered beverage formulations. Considering that higher andaliman proportions can enhance the solubility, pH, and overall product stability, the F3 formulation may be the most consumer-friendly and nutritionally beneficial option. These results highlight the potential of integrating andaliman and kuweni to create innovative powdered beverage formulations that are safe, stable, and functional, which cater to diverse consumer preferences while adhering to regulatory requirements (Baldissera et al., 2023). Future studies should focus on sensory evaluations, long-term stability, scalability, and consumer acceptability to further optimize these formulations for commercial success (Horacek et al., 2019).

The analysis of the physicochemical properties of the powdered beverage formulations reveals the critical role of ingredient selection in achieving the desired solubility levels and overall product quality. The findings emphasize the importance of optimizing formulation strategies to meet consumer demands for health-conscious, palatable, and stable beverage options. This research provides a solid foundation for future investigations aimed at enhancing the functionality and marketability of powdered beverages.

Total flavonoid content of andaliman- and kuweni-based powdered beverages

The increase in the flavonoid content with higher andaliman concentrations is a significant finding. Among the formulations, the F3 formulation exhibited the highest flavonoid concentration of 25.06 mg/g (P<0.05) (Fig. 1). This result is consistent with the findings of previous studies that have documented the phytochemical composition of andaliman, which includes a rich presence of flavonoids, tannins, saponins, and other bioactive compounds (Farida et al., 2021; Megawati et al., 2023; Simbolon et al., 2024). The increase in the flavonoid content can be attributed to the enhanced extraction efficiency of these compounds when higher andaliman concentrations are used, suggesting a concentration-dependent extraction process that maximizes the flavonoid yield.

Fig. 1.

Total flavonoid content of andaliman- and kuweni-based powdered beverages. Values are presented as mean±standard error from triplicate analysis (n=3). Different letters indicate significant differences (P<0.05). The error bars represent the standard error. F1, 25% andaliman:75% kuweni; F2, 50% andaliman:50% kuweni; F3, 75% andaliman:25% kuweni.

Flavonoids are known for their antioxidant properties, which are crucial for their anti-inflammatory and antidiabetic effects (Santoso et al., 2023; Zhang, 2023). The presence of flavonoids in andaliman has been linked to its potential therapeutic applications, particularly in the management of oxidative stress-related conditions. In the present study, the flavonoid content not only increases with the andaliman concentration but also suggests that specific fractions (e.g., F3) may contain higher concentrations of bioactive flavonoids, which contribute to the overall antioxidant capacity of the extract. This finding is consistent with the observation that individual flavonoids (e.g., quercetin) possess significant antioxidant activity (Silalahi et al., 2019).

Moreover, phytochemical analyses of andaliman fruit extracts revealed that they are rich in secondary metabolites, which are essential for the plant’s defense mechanisms and exhibit various pharmacological activities (Kholibrina and Aswandi, 2021). The presence of flavonoids in andaliman is particularly noteworthy as they play a pivotal role in plant physiology and contribute to the plant’s color, flavor, and pathogen resistance (Dalimunthe et al., 2024; Simbolon et al., 2024). The study’s findings underscore the importance of andaliman as a source of flavonoids, which can be harnessed for their health-promoting properties.

The specific flavonoid content in the F3 formulation, which is the highest among the tested concentrations, indicates that this fraction may contain a unique flavonoid profile that could be beneficial for further research and application in nutraceuticals (Santoso et al., 2023). The extraction process used in the present study, which involved ethanol as a solvent, effectively extracts flavonoids and other phenolic compounds, thereby enhancing their bioavailability and potential health benefits (Megawati et al., 2023). This method aligns with previous research that demonstrated the efficacy of ethanol extraction in maximizing flavonoid yields from various plant sources (Maleyki and Ismail, 2010).

Aside from the quantitative analysis of flavonoids, the qualitative aspects of these compounds needs to be considered. The specific types of flavonoids present in the F3 formulation could influence their biological activities and therapeutic potential. For example, quercetin and kaempferol have been extensively studied for their anti-inflammatory and antioxidant properties, making them valuable components in dietary supplements and functional foods (Silalahi et al., 2019). The identification of these compounds in andaliman could pave the way for the development of new health products that leverage this fruit’s unique phytochemical profile.

Furthermore, the increase in the flavonoid content with higher andaliman concentrations may also be influenced by the synergistic effects of other phytochemicals present in the extract. Moreover, the interaction between flavonoids and other bioactive compounds (e.g., tannins and saponins) could enhance the extract’s overall antioxidant capacity, providing a more comprehensive health benefit (Kholibrina and Aswandi, 2021; Simbolon et al., 2024). This synergistic effect is critical in understanding how andaliman can be used in functional foods and herbal medicine.

The implications of these findings extend beyond the laboratory as they suggest that andaliman could be incorporated into various food products to enhance their nutritional value. Because of its high flavonoid content, the potential use of andaliman as a natural preservative could also be explored, particularly in meat products where it has been shown to reduce bacterial growth and prolong shelf life (Marpaung et al., 2022; Patriani and Apsari, 2022). This application highlights andaliman’s versatility as a functional ingredient in food technology.

Antioxidant activity of andaliman- and kuweni-based powdered beverages

The antioxidant activity of powdered beverages formulated with varying ratios of andaliman and kuweni exhibited significant variations, as shown in Fig. 2. This highlights the role of andaliman as a potent source of antioxidants. In formulations with higher andaliman concentrations (e.g., F3 formulation), the antioxidant activity was markedly enhanced. This observation aligns with the findings of previous studies, which found that andaliman contains a rich phytochemical profile, including flavonoids, phenolic compounds, and other bioactive constituents known for their antioxidant properties (Natasutedja et al., 2020; Kholibrina and Aswandi, 2021; Zhang, 2023).

Fig. 2.

Antioxidant activity of andaliman- and kuweni-based powdered beverages. Values are presented as mean±standard error from triplicate analysis (n=3). Different letters indicate significant differences (P<0.05). The error bars represent the standard error. F1, 25% andaliman:75% kuweni; F2, 50% andaliman:50% kuweni; F3, 75% andaliman:25% kuweni.

The antioxidant efficacy of andaliman can be attributed to its high content of phenolic compounds, which can scavenge free radicals and chelate transition metals, thereby mitigating oxidative stress (Pieme et al., 2014; Hu et al., 2019). Moreover, the presence of flavonoids and other secondary metabolites in andaliman significantly contributes to its radical scavenging capacity as these compounds can donate electrons to free radicals, thereby effectively neutralizing them (Natasutedja et al., 2020; Anggraini et al., 2022). According to previous studies, the antioxidant activity of andaliman extracts is superior to that of many other plant sources, suggesting its potential application in functional foods and nutraceuticals (Farida et al., 2021; Anggraini et al., 2022; Zhang, 2023).

While also possessing antioxidant properties, kuweni may not exhibit the same level of efficacy as andaliman when used in lower concentrations. According to previous studies, the antioxidant activity of kuweni is influenced by its phenolic content, which, although beneficial, may not reach the potency observed in andaliman-rich formulations (Lestari et al., 2021; Nurhaliza et al., 2021). The comparative analysis of these two ingredients underscores the importance of formulation ratios in maximizing the antioxidant benefits. Formulations with a higher proportion of andaliman not only have enhanced overall antioxidant capacity but also provide additional health benefits associated with the consumption of phenolic-rich foods (Natasutedja et al., 2020; Kholibrina and Aswandi, 2021; Zhang, 2023).

The method used to assess the antioxidant activity, including DPPH assay, has been widely recognized for its reliability in evaluating the free radical scavenging ability of various compounds (Ilić et al., 2018; Dewana et al., 2022). In the context of beverages formulated with andaliman and kuweni, the results of DPPH assay indicated a clear correlation between the andaliman concentration and the observed antioxidant activity. Higher andaliman concentrations correlate with lower IC₅₀ values, suggesting a more potent antioxidant effect (Lestari et al., 2021; Dewana et al., 2022). This finding is consistent with those of previous studies that have established a direct relationship between phenolic content and antioxidant capacity in various plant extracts (Pieme et al., 2014; Hu et al., 2019; Anggraini et al., 2022).

Furthermore, the synergistic effects of combining andaliman with kuweni may play a role in enhancing the beverages’ overall antioxidant activity. According to previous studies, the interaction between different phytochemicals can lead to enhanced antioxidant effects, potentially through complementary mechanisms of action (Makanjuola et al., 2015; Yadav and Malpathak, 2016). The formulation of beverages that balance the antioxidant capacity of andaliman and kuweni could provide a more comprehensive approach to combating oxidative stress and promoting health (Kholibrina and Aswandi, 2021).

Aside from their antioxidant properties, andaliman and kuweni have been associated with various health benefits, including anti-inflammatory and antimicrobial effects, which further support their inclusion in functional food formulations (Farida et al., 2021). Incorporating these ingredients into traditional beverages not only enhances their nutritional profile but also aligns with the growing consumer demand for natural and health-promoting products (Anggraini et al., 2022).

The findings of the present study underscore the potential of andaliman and kuweni as valuable ingredients in developing functional beverages. Future studies should focus on exploring the specific mechanisms by which these ingredients exert their antioxidant effects and their potential interactions with other dietary components. In addition, investigating the bioavailability and metabolic pathways of the active compounds derived from andaliman and kuweni could provide further insights into their health benefits (Zhang, 2023).

Heavy metal and microbial contaminants of andaliman- and kuweni-based powdered beverages

The analysis of heavy metal and microbial contamination in andaliman- and kuweni-based powdered beverages revealed significant insights into the safety and quality of these products (Table 2). The findings show that all formulations (F1 to F3) comply with the maximum permissible limits for metal and microbial contaminants, suggesting that these beverages are safe for consumption. This compliance is critical because it aligns with international food safety standards, which emphasize the importance of monitoring contaminants in food products to protect public health (Pelegrín et al., 2020).

Table 2.

Heavy metal and microbial contaminants of andaliman- and kuweni-based powdered beverages

Characteristics	F1	F2	F3
Lead (mg/kg)	ND	ND	ND
Copper (mg/kg)	0.93±0.02a	0.82±0.01b	0.71±0.01c
Zinc (mg/kg)	0.29±0.00c	0.63±0.01a	0.55±0.01b
Tin (mg/kg)	ND	ND	ND
Arsenic (mg/kg)	ND	ND	ND
Total plate count (Log10 CFU/g)	2.58±0.02b	3.02±0.02a	1.98±0.08c
Coliform (MPN/g)	0.64±0.28	ND	ND

Values are presented as mean±standard error from triplicate analysis (n=3).

Different letters in a row indicate significant differences (P<0.05).

F1, 25% andaliman:75% kuweni; F2, 50% andaliman:50% kuweni; F3, 75% andaliman:25% kuweni; ND, not detected; CFU, colony-forming units; MPN, most probable number.

In terms of heavy metal contamination, the results showed that Pb, Sn, and As were undetectable across all formulations. This finding is particularly important because the presence of these toxic elements in food and beverages can pose serious health risks, including neurotoxicity and carcinogenic effects (Okafor et al., 2022). The Cu levels ranged from 0.71 mg/kg in F3 to 0.9 mg/kg in F1, which remained significantly below the maximum allowable limit of 2.0 mg/kg. Similarly, the Zn concentrations varied between 0.29 mg/kg in F1 and 0.63 mg/kg in F2, which are well below the safety threshold of 50 mg/kg (BSN, 1996). These findings underscore the effectiveness of controlled processing and the quality of ingredients used in these formulations, which is essential for ensuring consumer safety (Pelegrín et al., 2020).

The analysis of microbial contamination revealed that the TPC ranged from 1.98 log CFU/g in F3 to 3.02 log CFU/g in F2, which are within the acceptable limit of 3.48 log CFU/g (BSN, 1996). The presence of coliform bacteria was detected only in the F1 formulation, with levels of 0.64 MPN/g, which is within the permissible threshold of <3 MPN/g (BSN, 1996). The absence of coliform bacteria in the other formulations indicates that these batches have superior microbial control, which is crucial for maintaining the safety and quality of food products (Pelegrín et al., 2020; Jamilatun and Lukito, 2024).

Comparative analysis of the formulations revealed that F3, which contains 75% andaliman and 25% kuweni, exhibited the lowest microbial count, suggesting that higher andaliman concentrations may enhance the antimicrobial properties. This observation is consistent with the findings of previous studies that highlight the antimicrobial potential of various traditional ingredients, which can significantly reduce microbial loads in food products (Sharma, 2017). Conversely, the F1 formulation, which had lower andaliman concentrations, showed minor coliform contamination, potentially because of specific ingredient interactions or storage conditions. This finding indicates that the ingredient ratios and processing conditions in the formulation of traditional beverages need to be carefully considered (Jamilatun and Lukito, 2024).

The results of this study have significant implications for the production of traditional powdered beverages, highlighting the importance of ingredient ratios and processing controls in ensuring food safety. The low levels of heavy metals further support the products’ safety, aligning with international food safety standards (Pelegrín et al., 2020). Future studies should explore the mechanisms behind the antimicrobial effects of andaliman and optimize the formulation processes to enhance safety and functionality. This research reinforces the potential of traditional ingredients such as andaliman in developing safe and innovative functional beverages that not only meet safety standards but also provide health benefits (Fang et al., 2015; Pelegrín et al., 2020; Nemo and Bacha, 2021).

The analysis of metal and microbial contamination in andaliman- and kuweni-based powdered beverages demonstrates that these products are safe for consumption, with all formulations meeting the established safety standards. Moreover, the absence of toxic heavy metals and the controlled levels of microbial contaminants underscore the importance of the ingredients’ quality and processing methods in ensuring food safety. Continued research in this area will be essential to optimize traditional beverage formulations and enhance their safety and health benefits for consumers (Pelegrín et al., 2020).

In conclusion, this study highlights the potential of andaliman and kuweni as key ingredients in developing a functional powdered beverage with significant health benefits. The formulations demonstrated favorable physicochemical properties, including acceptable moisture content, enhanced solubility, and stable pH, which were influenced by the varying andaliman and kuweni proportions. The antioxidant activity, indicated by the total flavonoid content, showcased the synergistic potential of these ingredients. Furthermore, all formulations complied with the SNI for traditional powdered beverages, ensuring microbial safety and the absence of harmful heavy metals.

The findings of this study suggest that increasing the proportions of andaliman improves the solubility and pH, making these formulations consumer-friendly and nutritionally beneficial. This study bridges existing knowledge gaps by exploring the combined use of andaliman and kuweni in powdered beverage formulations and provides a foundation for future research on sensory attributes, long-term stability, and commercial scalability. Ultimately, the development of this powdered beverage contributes to the valorization of indigenous ingredients and the promotion of North Sumatra’s rich food heritage.