Physicochemical, Antioxidant, and Sensorial Properties of Xylopia aethiopica (Komba) Fruit on Mish Quality

Introduction

Herbal medicine (phytomedicine) represents a cornerstone of global alternative therapeutics.¹  Xylopia aethiopica (Dunal) A. Rich, a member of the Annonaceae family commonly known as African pepper, Ethiopian pepper, or “Komba” in Sudan, exemplifies this significance.² This tree, reaching heights exceeding 20 meters, inhabits diverse ecosystems including forests, riverbanks, and arid regions across Afric.³ Its distinctive fruit consists of slightly curved cylindrical pods that transition from green to brownish-black upon drying. Traditional medicinal practices in Sudan and Nigeria extensively utilize these fruits for treating respiratory ailments (coughs, bronchitis), gastrointestinal disorders (dysentery), edema, and as carminative or purgative agents.^4-8  This ethnopharmacological relevance stems from its complex phytochemical profile, documented to include alkaloids, saponins, sterols, tannins, phenols, flavonoids (e.g., rutin), cardiac glycosides, volatile and fixed oils,^9-13 complemented by vitamins (A-E), proteins, and essential minerals (copper, manganese, zinc)^12-15 While previous research has thoroughly characterized X. aethiopica‘s phytochemistry and in vitro bioactivities,^9-13,16-19 a critical knowledge gap persists regarding its functional application within traditional food systems, particularly fermented dairy products. This omission is notable for mish, a nutritionally significant Sudanese fermented milk consumed nationwide with regional variations in spicing intensity (e.g., “Haloom” among Bija tribes)²⁰. Traditionally prepared by adding soured milk to boiled milk with spices (black cumin, salt, fenugreek, red pepper) followed by 2-3 days’ fermentation,²⁰ mish presents an ideal vehicle for nutritional enhancement^21-25 and its biofunctional properties (antioxidative, anti-diabetic, and anti-inflammatory).^21,22,26-29 To address this research void, our study systematically investigated the novel fortification of mish with X. aethiopica fruit powder. Specifically, we examined the dose-dependent effects (0.0%, 0.5%, 1.0%, and 1.5% w/w) on key parameters: Chemical composition (total solids, protein, fat, fiber, ash, acidity), Mineral content (calcium, potassium, magnesium, iron), Bioactive properties (DPPH radical scavenging, total phenolics, flavonoids) and Sensory acceptability (appearance, texture, flavor, overall preference).

The goal was to establish optimal fortification levels that enhance nutritional and functional properties while maintaining organoleptic quality, thereby leveraging the documented significance of both X. aethiopica and mish within Sudanese dietary culture.

Material and Method

This study was conducted at Alzaeem Al-Azhari University’s food processing lab, which is part of the Department of Food Science and Technology, Faculty of Agriculture, Alzaeem Al-Azhari University, Sudan.

Materials

Yoghurt was sourced from Dal Dairy Factory (Capo), and spices (cumin, salt, and Xylopia aethiopica (Komba)) were procured from the central market in Khartoum, Sudan.

Experimental Design: A total of 6 kg of Capo yoghurt was divided into four treatments, each consisting of 1.25 kg per sample, with five replicates per treatment. The treatments included a control (0%), and three experimental groups with 0.5%, 1.0%, and 1.5% Xylopia aethiopica (Komba). Additionally, cumin (0.5%) and salt (1%) were added to each treatment.

Mish Manufacturing: The mish was prepared using Capo yoghurt produced a day prior. Cumin (0.5%) and salt (1%) were mixed into the yoghurt, which was then divided into four treatments of 1.25 kg each. Each treatment was further divided into six samples of 250 grams each. The levels of Xylopia aethiopica  (Komba) used were as follows: the control (0.0%), 0.5% (0.75 g per 250 g of mixture), 1.0% (1.5 g per 250 g), and 1.5% (2.25 g per 250 g). The samples were incubated at room temperature for 24 hours and, then stored at 5°C on the second day. Sensory evaluation and chemical analysis were conducted on the third day of manufacturing.

Chemical Analysis

The total solids content was measured using the drying oven method³⁰ protein content using the Kjeldahl method,³⁰ the fat content using the Gerber method and ash content according to (AOAC, 2000)30.  Finally, the mish samples were subjected to sensory evaluation by sixty panelists on the third day of production. The research protocol was previously approved by the Bioethics Committee of Qassim university (24-14-10) The panelists rated the samples on a five-point scale for appearance, texture, flavor, and overall acceptability. Each panelist was provided with water for rinsing between samples. The samples were coded before testing. The Statistical Package for Social Sciences (SPSS, 2004) was used to statistically evaluate the data. For statistical analysis, a completely randomized design was employed, and Duncan’s multiple range test was utilized to separate means at P < 0.001.

Results

Chemical Analysis of Mish Samples

Table (1) presents the total solids, protein, fat, ash, fiber, pH value, and titratable acidity of mish samples. The sample containing 1.5% Xylopia aethiopica  (Komba) exhibited the highest total solid content (17.0%), while the control sample (0.0%) had the lowest (13.80%). The other samples fell in between these values (P < 0.001).

Table 1: Impact of Xylopia aethiopica (Komba) fruit levels on physiological properties (%) of Mish.

*Mean ± SD. Rows with different superscript letters indicate significant differences (P < 0.001).

Minerals content of Mish samples

Table (2) presents the calcium, potassium, magnesium, and iron content in mish samples. The highest concentrations of calcium (980.22 mg/100g), potassium (482.61 mg/100g), magnesium (182.33 mg/100g), and iron (0.26 mg/100g) were found in the sample containing 1.5% Xylopia aethiopica (Komba). In contrast, the control sample had the lowest concentrations: calcium (895.60 mg/100g), potassium (455.35 mg/100g), magnesium (115.37 mg/100g), and iron (0.19 mg/100g). The other samples ranked in intermediate positions (P < 0.001).

Table 2: Impact of Xylopia aethiopica (Komba) fruit levels on minerals content (mg/100g) of mish.

*Mean ± SD. Rows with different superscript letters indicate significant differences (P < 0.001).

Sensory evaluation of Mish samples

Table (3) summarize the mean sensory attributes as evaluated by panelists for the four mish samples, focusing on appearance, texture, flavor, and overall acceptability.

Table 3: Impact of Xylopia aethiopica (Komba) fruit levels on organoleptic quality of mish.

Mean ± SD. Rows with different superscript letters indicate significant differences (P < 0.001).

Antioxidant activity of Mish samples in different concentration of Xylopia aethiopica fruit seeds

The data in Figure 1,2 and figure 3 shows the DPPH radical scavenging activity (mM Trolox equivalent ml-1), TFC (mg quercetin equivalent 100g-1 DW), and TPC (mg gallic acid equivalent 100g-1 DW) for each sample. Demonstrate how the concentration of Xylopia aethiopica fruits seeds affects the DPPH radical scavenging activity percentage, TFC, and TPC. The highest DPPH radical scavenging activity percentage, TFC, and TPC for mish samples with varying concentrations of Xylopia aethiopica  fruits and seeds (0.5, 1, and 1.5) was detected by all samples, but the sample containing 1.5% Xylopia aethiopica  (Komba) had the highest percentage (.84.4± 0.001-13.0 ±(0.001), (76.7±  0.1) and (39.9± 0.1)  while the control sample had the lowest (0.67.1± 0.001-.0.9±0.005), (30.5 ± 0.3) and ( 14.01± 0.1) DPPH radical scavenging activity percentage, TFC, and TPC respectively. The remaining samples were within the range of these values (P < 0.05). However, Onyenibe³¹  found DPPH % ug/ml) was 52.45± and Mojisola³²  found TFC, and TPC (mg/g) were (15.57± 0.20 and 11.71± 0.04) respectively.

Figure 1: Antioxidant activity (%) of X. aethiopica (Dose-dependent DPPH scavenging (84.4% at 1.5% Komba) Click here to view Figure

Figure 2: Total flavonoid compounds (mg/g) of X. aethiopica Click here to view Figure

Figure 3: Total phenolic compounds (mg/g) of X. aethiopica Click here to view Figure

Discussion

Chemical Analysis of Mish Samples

The high total solids content may be attributed to fermentation, flavoring addition, and the concentration and conversion of raw milk from liquid to gel form.³³

The highest protein content (5.95%) was also observed in the sample with 1.5% Xylopia aethiopica  (Komba), with the control sample having the lowest (4.75%). The remaining samples had intermediate rankings (P < 0.001). These results were higher than those reported by ( El-Zubeir et al, 2005),)³⁴ who found the protein content of mish to be 5.09±2.80 and (Ajibade etal,2025)³⁵ The increased protein content in the mish samples may be due to the varying levels of Xylopia aethiopica (Komba) used during production.

Similarly, the highest fat content (5.05%) was found in the sample with 1.5% Xylopia aethiopica  (Komba), while the control sample had the lowest (3.94%). The remaining samples had intermediate rankings (P < 0.001).  These values were lower than those reported by (Nassib & El-Gendy, 1974)³⁶ and ( El-Zubeir et al, 2005 and Ajibade et al,2025)^34,35 for mish.

The Sudanese Standards and Metrology Organization specifies that the fat content in mish should not exceed 3%, highlighting a discrepancy between the findings of this study and the recommended values.³⁷ The sample with 1.5% Xylopia aethiopica  (Komba) had the greatest ash concentration (2.77%), while the control sample (0.0%) had the lowest ash content (1.25%), with other samples falling in between (P < 0.001). These results align with El Zubeir,³⁴who reported an ash content of 1.26% but  are higher than those reported by El Erian.³⁸

The highest fiber content (1.26%) was also found in the sample with 1.5% Xylopia aethiopica  (Komba), whereas the control sample had no detectable fiber (0.00%). There were several samples that placed in the middle (P < 0.001). These results are lower than those reported by Osabor,^35,39 who found a crude fiber content of 3.87% in Xylopia aethiopica (Komba) fruit, which also revealed significant mineral concentrations.

The control sample (0.0% Xylopia aethiopica  (Komba)) had the highest pH value (5.80), while the lowest pH value (5.05) was found in the sample with 1.5% Xylopia aethiopica  (Komba), with other samples ranking in between (P < 0.001). These pH values are higher than those reported by Osman,⁴⁰ but similar to those reported by Nassib and El-Gendy.³⁶ The increase in acidity, resulting in lower pH values, is attributed to the fermentation process, which increases acid concentration.

The highest titratable acidity (2.06%) was observed in the sample with 1.5% Xylopia aethiopica  (Komba), while the control sample had the lowest (1.70%), with other samples ranking in intermediate positions (P < 0.001). These findings surpass those of,³⁴ who discovered that the titratable acidity of mish samples was 1.24% ± 0.41. The increase in acidity is primarily due to the proliferation of lactic acid bacteria, which convert lactose into lactic acid.^41-45 

Minerals content of Mish samples

The results indicate a higher concentration of calcium, which is necessary for the healthy development and upkeep of teeth and bones, as well as for nerve transmission.⁴⁶ The daily recommended intake of calcium is 360-400 mg/day for both children and adults. However, these findings are lower than those reported by³⁹ who found calcium levels of 2.13 mg/100g in Xylopia aethiopica fruit.

This study’s potassium content was also less than what was reported by Osabor,³⁹ who found potassium levels of 4.80 mg/100g in Xylopia aethiopica  fruit. Potassium ions help regulate the pH of intracellular fluids and maintain osmotic pressure within cells.⁴⁷

Similarly, the magnesium content was lower than that reported by Osabor,³⁹ who found magnesium levels of 2.40 mg/100g in Xylopia aethiopica  fruit. Magnesium is crucial for various biophysiological processes, including enzyme activation and DNA structure stabilization.⁴⁸

The iron content in this study was lower than that reported by John-Dewole,¹¹ who found iron levels of 0.690 mg/100g in Xylopia aethiopica  fruit through spectroscopic analysis of rare metals. 

Sensory evaluation of Mish samples

The highest ratings for overall acceptability (4.8), texture (4.8), flavor (4.9), and appearance (4.7) were given to the sample that contained 1.0% Xylopia aethiopica (Komba) fruit. On the other hand, the sample containing 0.0% Xylopia aethiopica (Komba) scored the lowest in terms of general acceptability (4.2), texture (4.4), flavor (4.1), and appearance (4.2). The other samples ranked in intermediate positions (P < 0.001). 

Antioxidant activity of Mish samples in different concentration of Xylopia aethiopica fruit seeds

The rich flavonoids and phenolic compounds found in X. aethiopica fruit position it as a natural remedy for various conditions including cancer, heart disease, atherosclerosis, and peptic ulcers prevention. These compounds, notably flavonoids, act as potent antioxidants crucial in combatting free radicals and preventing degenerative diseases such as heart conditions.⁴⁹ Moreover, flavonoids play a significant role in regulating the cell cycle, inducing apoptosis, and inhibiting the growth of cancer cells.⁵⁰ Their multifaceted functions as anti-proliferative agents, estrogen modulators, and antioxidants offer protection against cancer and atherosclerosis. Additionally, phenolic compounds, recognized for their antioxidant properties, exhibit anti-inflammatory, anticancer, and atherosclerotic effect. ^51-53 Studies have also highlighted the potential of plant extracts rich in flavonoids in the prevention of peptic ulcers.^15,54 

Conclusion

In the study, it was investigated that the effects of different levels of Xylopia aethiopica (Komba) on the quality of mish, a vital component of the Sudanese diet. The study utilized yoghurt from the Dal Dairy Factory and Xylopia aethiopica fruits from Khartoum State, Sudan. These fruits were air-dried and manually ground to create four distinct treatment groups: a control group with 0.0% Xylopia aethiopica and experimental groups with concentrations of 0.5%, 1.0%, and 1.5%. Following incubation and storage, thorough assessments were conducted on the third day, including analyses of chemical composition, antioxidant activity (DPPH radical scavenging activity percentage, TFC, TPC), and sensory evaluations. The results revealed that the mish sample containing 1.5% Xylopia aethiopica exhibited superior characteristics, with higher levels of total solids, protein, fat, fiber, and acidity compared to other samples. Additionally, it showed elevated levels of calcium, potassium, magnesium, and iron. In sensory evaluation, the mish sample with 1.0% Xylopia aethiopica was favored for its exceptional appearance, texture, flavor, and overall acceptability. This study highlights the significant improvement in nutritional parameters and antioxidant activity in mish through the inclusion of Xylopia aethiopica. It emphasizes the potential of Xylopia aethiopica as a functional ingredient with nutritional and antioxidant benefits, suggesting its potential for enhancing mish quality in the food industry and potentially in medicinal applications. Future studies should explore encapsulation techniques to optimize sensory acceptance of higher-concentration Xylopia aethiopica-fortified mish while preserving its enhanced nutraceutical properties for commercial scalability.

Acknowledgement

The Researchers would like to thank the Deanship of Graduate Studies and Scientific Research at Qassim University for financial support (QU-APC-2025)

Funding Sources

The author(s) received financial support for the publication of this article. 

Conflict of Interest

The authors declare no conflict of interest.

Data Availability Statement

The article contains the information that was utilized to support the study’s conclusions.

Ethics Statement

The research protocol was previously approved by the Bioethics Committee of Qassim University (24-14-10).

Informed Consent Statement

This study did not involve human participants, and therefore, informed consent was not required. 

Clinical Trial Registration

This research does not involve any clinical trials.

Permission to Reproduce Material from Other Sources

The manuscript did not contain any materials such as figures, tables, or text excerpts that have been previously published elsewhere.

Author Contributions

• E.Ahmed: Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Writing – original draft, Writing – review & editing  
• Kamal Awad Abdel Razig: Conceptualization, Data curation, Investigation, Methodology, Supervision, Resources, Writing – original draft, Writing – review & editing.
• Mohamed G.E. Gadallah: Conceptualization, Formal analysis, Resources, Writing – review & editing
• Ghadah S. Alrebdi : Conceptualization, Formal analysis, Resources, Writing – review & editing
• Ammar AL-Farga: Conceptualization, Formal analysis, Resources, Writing – review & editing
• Islam Ragab: Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Writing – original draft, Writing – review & editing

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