Influence of Fermentation in Water and Sugarcane Juice on Nutritional Composition of Azanza garckeana Fruit

Influence of Fermentation in Water and Sugarcane Juice on Nutritional Composition of Azanza garckeana Fruit | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Influence of Fermentation in Water and Sugarcane Juice on Nutritional Composition of Azanza garckeana Fruit Mansurat B. Falana, Quadri O. Nurudeen, Hafisat Tijani, Muhammed R. Asinmi, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8627658/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 10 You are reading this latest preprint version Abstract Traditionally consumed as food and medicine, the impact of fruit fermentation of Azanza garckeana (F. Hoffm.) Exell & Hillc. in distilled water (AGW) and sugarcane juice (AGS) for 3 and 5 days, and the nutritional composition was examined. The moisture level of AGW and AGS decreased (60.05% and 59.04% to 54.54% and 57.83%). Protein reduced from 3.44% (AGW) and 5.81% (AGS) to 2.81% and 2.42%, respectively. Lipid increased from 5.22 (AGW) to 6.44%, but decreased from 3.11 (AGS) to 2.72%. Crude fiber slightly reduced (25.34–23.57%) in AGW and increased (21.35–22.87%) in AGS. Carbohydrate increased from 0.79–8.33% to 7.36–14.15% in AGW and AGS, respectively. Liquid Chromatography-Mass Spectrometry (LC-MS) identified 16 amino acids; glutamic acid, arginine, and cysteine were abundant, while tyrosine was sparse. Gas Chromatography-Mass Spectrometry (GC-MS) identified higher vitamins C, B9, and A on day 5 and in AGS. X-ray fluorescence (XRF) revealed macro-minerals (O, Mg, P, Cl, K, Ca, S), micro-minerals (Cu, Zn, Cr, Mn, Fe, V), and trace elements (Si, Ag). Cl and O decreased (60–42 and 12.1–21) in AGW, but increased (39–52) in AGS. Fermented A. garckeana is an affordable, nutritious, and sustainable bioresource that addresses zero hunger, poverty alleviation, good health, and economic empowerment. Azanza garckeana Fermentation Nutritional Composition Amino acids Sustainable Development Goals Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 1. Introduction Plants are a source of minerals, vitamins, protein, and key amino acids that are vital for the body's metabolism (Osum et al., 2013 ). Different parts of edible plants, naturally growing in the forest or grown by residents, are consumed as food and offer numerous medicinal values (Nazarudeen, 2010 ; Nkafamiya et al., 2016 ). The preparation of plants and the preservation of their bioactive components for medicinal purposes have been major challenges. Hence, the significance of fermentation, a microbial and enzyme-catalyzed process that improves food shelf life, boosts nutritional and bioactive qualities (Di Cagno et al., 2013 ). Azanza garckeana (F. Hoffm.) Exell & Hillc., also known as “Goron Tula” or “African chewing gum,” is a popular wild edible plant of the Malvaceae. It is commonly found in the Tula community of Kaltungo LGA, Gombe State, Nigeria. It is a commercially viable crop that is consumed locally (Ochokwu et al., 2015 ). It was also reported to be found in other parts of West Africa, including Ghana and Senegal, where it is used for many purposes (Bukar et al., 2020 ; Bioltif et al., 2020 ). A. garckeana is an important source of vitamins, minerals, carbohydrates, calcium, salt, phosphorus, iron, and ascorbic acid, making it an essential nutritional supplement in areas where it is grown (Sulieman, 2019 ; Yusuf et al., 2020 ; Nurudeen et al., 2024 ). According to previous scientific reports, A. garckeana is a nutrient-dense fruit rich in fiber, carbohydrates, protein, and ash, but relatively low in fat (Abass and Ahmed, 2017 ). The presence of various amino acids, including aspartic acid, alanine, arginine, cysteine, glycine, glutamic acid, proline, serine, tyrosine, and lysine, in the fruits and leaves of A. garckeana was also documented (Nkafamiya et al., 2016 ). Essential amino acids (histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, and valine), suggesting their potential in alleviating protein-energy malnutrition (Bioltif et al. ,2020). In addition, the fruit contains significant amounts of vitamins A, B1, B2, C, and E (Nkafamiya et al., 2016 ; Danbature et al., 2015 ; Michael et al., 2015 ), which further highlights its nutritional and medicinal value. Despite the existing reports on A. garckeana , information is lacking on its fermented form. Hence, this study explored the proximate and nutritional composition of A. garckeana fruits fermented in distilled water and sugarcane juice, respectively. 2. Materials and Methods 2.1 Procurement and processing of raw materials Sugarcane and matured fresh fruits of Azanza garckeana of Nigerian origin were purchased from a local Market in Ilorin, Nigeria. The Azanza garckeana fruits, previously identified by local herb sellers, were further authenticated by a taxonomist, Mr. Bolu Ajayi, at the Herbarium Unit, Department of Plant Biology, University of Ilorin, Nigeria, where a voucher specimen was deposited under the accession number UILH/001/1828/2025. The ripe fruits were sorted to remove stones, cut into smaller sizes, and rinsed quickly in distilled water. 2.2 Processing and Fermentation Adopting the method described by Falana et al. ( 2024 ), fermentation of Azanza garckeana was achieved in two sterilized Mason jars. Distilled water (1560mL) and 40g of iodized salt were added to a jar, while the second jar contained sugarcane juice in place of distilled water. The prepared fruit of A. garckeana (400g) was added to each jar in a ratio of 1:4 w/v (prepared fruit sample to the brine solvent). Iodized salt was added to discourage the growth of spoilage organisms and inhibit potential pectinolytic and proteolytic enzymes that may aid putrefaction. The airlock was tightly covered to facilitate spontaneous fermentation, and the setup was kept at room temperature (28 ± 2°C). However, the airlock was loosened after 24 hours to release carbon dioxide. The jar was then stirred to encourage natural microbial growth. The medium was allowed to ferment for 5 days. At day 3, some of the sample was collected and filtered using a 0.4 mm membrane. The sample was refrigerated at -4 o C for analysis (Guarner and Schaafsma, 1998 ). 2.3 Quantification of Proximate Composition Proximate composition of the fermented samples was determined as described by the Association of Official Analytical Chemists (AOAC, 2012). 2.4 Amino acid content (LC-MS) The fermented A. garckeana were added into water containing 13C, 15N-labeled algal amino acid mix (Cambridge Isotope Laboratories, Inc., USA) at a concentration of 10 g/mL. The amino acids in the diluted A. garckeana sample were subsequently analyzed in LC-MS/MS Sciex QTRAP 6500 + triple-quadrupole-trap MS/MS (Jander et al., 2004 ; Vadassery et al., 2014 ; Crocoll et al., 2016 ). Also, the amino acid was separated with a Zorbax Eclipse XDB-C18 column (50 × 4.6 mm, 1.8 µm, Agilent Technologies). The mobile phase consisted of two solvents, A (water with 0.1% formic acid) and B (acetonitrile with 0.1% formic acid). The flow rate was maintained at 1.1 mL per min with the elution profile ran as follows: 0–1 min, 3% B; 1-3.8 min, 3–50% B; 3.8–3.9 min 50–100% B, 3.9-5 min, 100%B; 5-5.1 min, 100-3% B, 5.1-7 min, 100% B. The column was held at a constant temperature (27°C). The analyte parent ion and product ion were monitored closely using the mass spectrometer in positive ionization mode (MRM modus) for detection Jander et al., 2004 ). Both Q1 and Q3 quadrupoles were maintained at unit resolution. Furthermore, Analyst 1.5 software (Sciex) was used for data acquisition and processing. Using the method described by Muni et al. ( 2023 ), individual amino acids in the sample were quantified with 13C and 15N-labeled amino acids, respectively. 2.5 GC-MS Analysis of Vitamins The samples (AGW and AGS) were analyzed using GC-MS to identify and quantify the vitamins present. The analysis was performed with an Agilent Technologies 7890 GC system coupled to a 5975 Mass Spectrometry detector. Separation was achieved using an Agilent HP-5MS capillary column (30 m × 0.320 mm × 0.25 µm film thickness). Helium served as the carrier gas at a constant flow rate of 0.5 mL/min, and an injection volume of 1 µL was introduced into the system. The oven temperature was initially set at 80°C for 2 minutes, then increased at 10°C per minute to a final temperature of 240°C for 6 minutes. The total run time was 90 minutes. Electron impact ionization was conducted at 70 eV. Vitamins identification and quantification were based on Total Ion Chromatograms (TIC). The resulting spectra were compared with reference spectra in the National Institute of Standards and Technology (NIST) library (Gaithersburg, Maryland, USA) for accurate compound identification (Momodu et al., 2022 ). 2.6 Mineral Content Analysis (XRF) Elemental minerals of the fermented samples were determined by EDXRF analysis using an ARL (Yagi et al., 2013 ). The Quant’X system is equipped with a cellulose X-ray filter and a Rh-anode X-ray tube. The system operated within a voltage range of 8–12 kV and a current intensity of 0.32–0.34 mA. A Si (Li) detector cooled with liquid nitrogen was used to detect the emitted X-rays. Each sample was analyzed for 120 seconds, with an average dead time of approximately 50%. Data processing was performed using WINTRACE software version 4.1, build 9. All analyses were conducted in triplicate for each plant sample to ensure accuracy and reproducibility. 2.7 Data Analysis The results obtained in triplicate were expressed as mean values and standard deviation (SD). All analyses were performed using Microsoft Excel software 3. Results and Discussion 3.1 Proximate Composition of A. garckeana This study revealed the highest moisture and carbohydrate contents in AGS at day 5, but the lowest in AGW at day 3 (Table 1 ). Protein and Ash content reduced with increasing fermentation duration for both samples. Lipid content was highest in AGW (5 days) but lowest in AGS (5 days). The increased moisture observed in the current study may indicate accelerated fermentation due to the growth of desirable microorganisms and the influence of their metabolites, such as acids and alcohols, as reported in similar fruit fermentations (Di Cagno et al., 2013 ). Driven by microbial enzymes, the increased carbohydrate in this study may indicate the presence of resistant starches. During fermentation, starches are converted into simpler and digestible forms like acetic acid and lactic acid. Acetic acid and lactic acid lower pH, promote the shelf life of the sample, and enhance the digestibility of foods, positioning fermented A. garckeana as a valuable energy source for populations at risk of protein–energy malnutrition (Nkafamiya et al., 2016 ). Hence, supports SDG 2 on Zero Hunger. The decrease in protein content with time corroborates earlier findings by Falana et al. ( 2024 ), who explained that fermenting microorganisms may have broken down the protein into amino acids and peptides. The increased lipid content may mean microbial synthesis and accumulation of lipid metabolites, consistent with observations by Falana et al. ( 2024 ). Table 1 Proximate Composition of the Two Fermented Varieties of A. garckeana Mean ± SEM Parameters AGW (3 Days) AGW (5 Days) AGS (3 Days) AGS (5 Days) Moisture 60.05 ± 0.005 54.54 ± 0.010 59.04 ± 0.010 57.83 ± 0.040 Protein 3.44 ± 0.0004 2.81 ± 0.000 5.81 ± 0.0005 2.42 ± 0.0015 Lipid 5.22 ± 0.005 6.44 ± 0.000 3.11 ± 0.005 2.72 ± 0.000 Crude fiber 25.34 ± 0.005 23.57 ± 0.0005 21.35 ± 0.000 22.87 ± 0.000 Ash content 5.21 ± 0.001 4.33 ± 0.001 3.33 ± 0.004 3.81 ± 0.001 Carbohydrate 0.79 ± 0.000 8.31 ± 0.0005 7.36 ± 0.004 14.15 ± 0.004 Key: AGW : A. garckeana fermented in water, AGS : A. garckeana fermented in sugarcane juice 3.2 LC-MS Mineral Composition of A. garckeana Seventeen amino acids were identified in samples (Fig. 1 ), with glutamic acid being the most abundant and tyrosine the least in both fermented samples. Such amino acids play crucial physiological roles in energy metabolism, neurotransmission, tissue repair, and collagen synthesis, suggesting that fermented A. garckeana may have cognitive and physiological health-boosting properties. This is consistent with previous compositional studies on A. garckeana (Nkafamiya et al., 2016 ; Bioltif et al., 2020 ). The sugar content of sugarcane juice may be bio-enhancing, leading to the optimized nutrient conversion and synthesis of essential compounds in the sample (Di Cagno et al., 2013 ). The increased proline and leucine with fermentation time indicate extensive proteolysis and amino acid liberation, enhanced digestibility, and metabolic utilization (Nkafamiya et al., 2016 ). 3.3 GC-MS Vitamins Composition of A. garckeana Notably, vitamins A, B9, and C increased significantly in sugarcane-fermented samples after five days (Figs. 2 – 6 ). This reinforces the role of sugar-rich substrates in microbial synthesis of bioactive compounds (Falana et al., 2024 ). Higher mineral concentrations, including calcium, potassium, and chlorine, were recorded in sugarcane-fermented samples, all of which are critical for muscle function, nerve transmission, and electrolyte regulation (Ochokwu et al., 2015 ). In line with our study, Sulieman ( 2019 ) asserted that fermentation improves the nutritional and sensory properties of local fruits. 3.4 Mineral Composition of A. garckeana by X-ray fluorescence Seven macro minerals (Oxygen, magnesium, phosphorus, chlorine, potassium, calcium, and sulfur), six microminerals (Copper, zinc, chromium, manganese, iron, and vanadium), and two other biological essential elements (silicon and silver) were identified in the fermented samples (Table 2 ). The elevated potassium and calcium concentrations observed in sugarcane-fermented samples align with prior studies by Ochokwu et al. ( 2015 ) and Momodu et al. ( 2022 ), which highlighted the mineral richness of A. garckeana . The presence of biologically useful trace elements such as iron and zinc may be linked to hemoglobin synthesis, enzymatic regulation, and immune defense (Abass and Ahmed, 2017 ). Also, trace elements silicon and vanadium, which are associated with bone metabolism and catalytic enzyme functions (Yagi et al., 2013 ), emphasize the potential of the fermented samples as functional foods for well-being and support SDG 3 on Good Health and Well-being. Table 2 Comparative Elemental Mineral Content of the Two Fermented Varieties of A. garckeana S/n Element Symbol AFW 3 Days AFW 5 Days AFS 3 days AFS 5 Days 1 Oxygen O 12.1 21.4 25.2 15 2 Magnesium Mg 0.1 1.4 0.1 0.01 3 Copper Cu 1.3 0.5 0.8 0.8 4 Zinc Zn 0.4 0.1 0.1 0.1 6 Silicon Si 3.3 3.0 7.5 5.0 7 Phosphorus P 0.3 0.2 0.4 0.4 8 Chlorine Cl 60 42 39 52 9 Potassium K 15 11 8.5 16 10 Calcium Ca 2.2 1.5 3.4 5.1 11 Vanadium V 0.1 0.01 0.1 0.01 12 Chromium Cr 0.1 0.1 0.1 0.1 13 Manganese Mn 0.1 0.1 0.1 0.1 14 Iron Fe 0.7 0.2 0.9 0.6 14 Sulfur S 0.1 0.1 0.01 0.01 16 Silver Ag 0.4 0.2 0.3 0.1 Key: AGW : A. garckeana fermented in water, AGS : A. garckeana fermented in sugarcane juice 4. Conclusion This study demonstrates that medium and duration of fermentation influence the nutritional composition of Azanza garckeana . Sugarcane juice and extended duration optimized the levels of vitamins (A, B9, and C), carbohydrates, and lipids while promoting the release of amino acids such as glutamic acid, leucine, and proline, which contribute to improved digestibility and physiological performance. Vital macro- and microelements—particularly potassium, calcium, zinc, and iron, which support metabolic, immune, and hematological health, were present. These results affirm that fermentation not only preserves A. garckeana but also transforms it into a functional food with enhanced nutritional and health-promoting potential. Thus, beyond the nutritional relevance of fermentation, it serves as a bio-enhancing and sustainable process that promotes microbial vitamins and mineral synthesis, and contributes to the realization of key Sustainable Development Goals (SDGs), especially SDG 2 (Zero Hunger), SDG 3 (Good Health and Well-being), and SDG 12 (Responsible Consumption and Production). Declarations Data Availability Statement All data generated or analysed during this study are included in this published article [and its supplementary information files]. Competing interests There are no competing interests. All authors certify that they have no affiliations with or involvement in any organization or entity with any financial interest or non-financial interest in the subject matter or materials discussed in this manuscript. Funding The authors have not received any funding for this study. Authors' contributions MBF developed the review protocol and study design. MBF and HT drafted the manuscript; MRB and MAD were responsible for data analysis and interpretation. MBF and QON edited the manuscript and contributed to data analysis and interpretation. All authors read and approved the final manuscript. Acknowledgements Not applicable Clinical trial Not applicable Ethics, Consent to Participate, and Consent to Publish declarations The fruits of Azanza garckeana (F. 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Falana","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA5UlEQVRIiWNgGAWjYBACAwbmBiDFBmIzPmBgOECMFkaQFj4Qm9mAFC1yIDabBFFazNkbWzf83GEmZ3C891k1T80dOX4G5oePbuDRYtlzsO1m75k0Y4Mzx81u8xx7ZizZwGZsnIPPYTcS227wth1LnDkjje02D9vhxA0HeNikCWm5+bftf/3M+c/Yinn+EanlNm8bWwK/BBsbM28bEVpAfrkt28Zm2M+Txiw5t++wsWQzAb+Yszcfu/m2jU2ejf0Y44c33w7L8bM3P3yMTwsKYOIBkczEKgcBxh+kqB4Fo2AUjIIRAwDpLU8bKuE5JQAAAABJRU5ErkJggg==","orcid":"","institution":"Al-Hikmah University","correspondingAuthor":true,"prefix":"","firstName":"Mansurat","middleName":"B.","lastName":"Falana","suffix":""},{"id":588942750,"identity":"6fc68887-5ed4-463c-87ad-8c270cae6d98","order_by":1,"name":"Quadri O. Nurudeen","email":"","orcid":"","institution":"Al-Hikmah University","correspondingAuthor":false,"prefix":"","firstName":"Quadri","middleName":"O.","lastName":"Nurudeen","suffix":""},{"id":588942751,"identity":"a0f8a5c7-a635-48f4-a25f-fd820ae9a67f","order_by":2,"name":"Hafisat Tijani","email":"","orcid":"","institution":"Al-Hikmah University","correspondingAuthor":false,"prefix":"","firstName":"Hafisat","middleName":"","lastName":"Tijani","suffix":""},{"id":588942753,"identity":"2cbff097-a571-41a9-a685-f0556f6c0126","order_by":3,"name":"Muhammed R. Asinmi","email":"","orcid":"","institution":"Al-Hikmah University","correspondingAuthor":false,"prefix":"","firstName":"Muhammed","middleName":"R.","lastName":"Asinmi","suffix":""},{"id":588942754,"identity":"b69962d4-907f-4d9c-bb76-5b47bf42289a","order_by":4,"name":"Muhammad A. Dikwa","email":"","orcid":"","institution":"Federal University Dutse","correspondingAuthor":false,"prefix":"","firstName":"Muhammad","middleName":"A.","lastName":"Dikwa","suffix":""}],"badges":[],"createdAt":"2026-01-17 17:53:14","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8627658/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8627658/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":102623817,"identity":"4c517ab7-4254-4330-ad96-189c3353a428","added_by":"auto","created_at":"2026-02-13 17:18:08","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":45724,"visible":true,"origin":"","legend":"\u003cp\u003eComparison of the Amino Acid Composition of the Two Varieties of Fermented \u003cem\u003eA. garckeana\u003c/em\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-8627658/v1/1564a36f95f8fba0e4d01775.png"},{"id":102623821,"identity":"4859cacc-dd4f-46e7-954c-d59b19a480aa","added_by":"auto","created_at":"2026-02-13 17:18:08","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":82787,"visible":true,"origin":"","legend":"\u003cp\u003eHPLC Chromatogram of Vitamins in 3-day Fermented \u003cem\u003eA. garckeana\u003c/em\u003e in Sugarcane Juice\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-8627658/v1/2d87e1421657ce67a2c4b6c2.png"},{"id":102623818,"identity":"eea7c8a5-b157-4165-beb4-e154690485bf","added_by":"auto","created_at":"2026-02-13 17:18:08","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":67756,"visible":true,"origin":"","legend":"\u003cp\u003eHPLC Chromatogram of Vitamins in 5-day Fermented \u003cem\u003eA. garckeana\u003c/em\u003e in Sugarcane Juice\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-8627658/v1/c250b64ec41ea89c04660d30.png"},{"id":102748087,"identity":"0fd296bd-809c-4fd0-8891-8d7ace2ca1a5","added_by":"auto","created_at":"2026-02-16 09:05:54","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":70462,"visible":true,"origin":"","legend":"\u003cp\u003eHPLC Chromatogram of Vitamins in 3-day Fermented \u003cem\u003eA. garckeana\u003c/em\u003e in Distilled Water\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-8627658/v1/6d0610bf68f0872e2fe5fe8e.png"},{"id":102747861,"identity":"bae842ec-3479-4d55-995a-211707a25851","added_by":"auto","created_at":"2026-02-16 09:05:30","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":69281,"visible":true,"origin":"","legend":"\u003cp\u003eHPLC Chromatogram of Vitamins in 5-day Fermented \u003cem\u003eA. garckeana\u003c/em\u003e in Distilled Water\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-8627658/v1/7a72b45fde26ff1b098653e3.png"},{"id":102623822,"identity":"c0acf759-a647-4b09-8107-72ba2c1189c5","added_by":"auto","created_at":"2026-02-13 17:18:08","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":23212,"visible":true,"origin":"","legend":"\u003cp\u003eComparison of Vitamin Composition of the Two Varieties of Fermented \u003cem\u003eA. garckeana\u003c/em\u003e\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-8627658/v1/31ac13ba4e3c1198837b0fce.png"},{"id":102750984,"identity":"2bef13c2-3c65-4f96-a84d-9d06684a2e22","added_by":"auto","created_at":"2026-02-16 09:23:01","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1111099,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8627658/v1/5b1a17f1-df4d-4e24-9697-13abb2bb4c80.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Influence of Fermentation in Water and Sugarcane Juice on Nutritional Composition of Azanza garckeana Fruit","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003ePlants are a source of minerals, vitamins, protein, and key amino acids that are vital for the body's metabolism (Osum et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Different parts of edible plants, naturally growing in the forest or grown by residents, are consumed as food and offer numerous medicinal values (Nazarudeen, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Nkafamiya et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). The preparation of plants and the preservation of their bioactive components for medicinal purposes have been major challenges. Hence, the significance of fermentation, a microbial and enzyme-catalyzed process that improves food shelf life, boosts nutritional and bioactive qualities (Di Cagno et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2013\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cem\u003eAzanza garckeana\u003c/em\u003e (F. Hoffm.) Exell \u0026amp; Hillc., also known as \u0026ldquo;Goron Tula\u0026rdquo; or \u0026ldquo;African chewing gum,\u0026rdquo; is a popular wild edible plant of the Malvaceae. It is commonly found in the Tula community of Kaltungo LGA, Gombe State, Nigeria. It is a commercially viable crop that is consumed locally (Ochokwu et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). It was also reported to be found in other parts of West Africa, including Ghana and Senegal, where it is used for many purposes (Bukar et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Bioltif et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). \u003cem\u003eA. garckeana\u003c/em\u003e is an important source of vitamins, minerals, carbohydrates, calcium, salt, phosphorus, iron, and ascorbic acid, making it an essential nutritional supplement in areas where it is grown (Sulieman, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Yusuf et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Nurudeen et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAccording to previous scientific reports, \u003cem\u003eA. garckeana\u003c/em\u003e is a nutrient-dense fruit rich in fiber, carbohydrates, protein, and ash, but relatively low in fat (Abass and Ahmed, \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The presence of various amino acids, including aspartic acid, alanine, arginine, cysteine, glycine, glutamic acid, proline, serine, tyrosine, and lysine, in the fruits and leaves of \u003cem\u003eA. garckeana\u003c/em\u003e was also documented (Nkafamiya et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Essential amino acids (histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, and valine), suggesting their potential in alleviating protein-energy malnutrition (Bioltif \u003cem\u003eet al.\u003c/em\u003e,2020). In addition, the fruit contains significant amounts of vitamins A, B1, B2, C, and E (Nkafamiya et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Danbature et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Michael et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), which further highlights its nutritional and medicinal value.\u003c/p\u003e \u003cp\u003eDespite the existing reports on \u003cem\u003eA. garckeana\u003c/em\u003e, information is lacking on its fermented form. Hence, this study explored the proximate and nutritional composition of \u003cem\u003eA. garckeana\u003c/em\u003e fruits fermented in distilled water and sugarcane juice, respectively.\u003c/p\u003e"},{"header":"2. Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Procurement and processing of raw materials\u003c/h2\u003e \u003cp\u003eSugarcane and matured fresh fruits of \u003cem\u003eAzanza garckeana\u003c/em\u003e of Nigerian origin were purchased from a local Market in Ilorin, Nigeria. The \u003cem\u003eAzanza garckeana\u003c/em\u003e fruits, previously identified by local herb sellers, were further authenticated by a taxonomist, Mr. Bolu Ajayi, at the Herbarium Unit, Department of Plant Biology, University of Ilorin, Nigeria, where a voucher specimen was deposited under the accession number UILH/001/1828/2025. The ripe fruits were sorted to remove stones, cut into smaller sizes, and rinsed quickly in distilled water.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Processing and Fermentation\u003c/h2\u003e \u003cp\u003eAdopting the method described by Falana et al. (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), fermentation of \u003cem\u003eAzanza garckeana\u003c/em\u003e was achieved in two sterilized Mason jars. Distilled water (1560mL) and 40g of iodized salt were added to a jar, while the second jar contained sugarcane juice in place of distilled water. The prepared fruit of \u003cem\u003eA. garckeana\u003c/em\u003e (400g) was added to each jar in a ratio of 1:4 w/v (prepared fruit sample to the brine solvent). Iodized salt was added to discourage the growth of spoilage organisms and inhibit potential pectinolytic and proteolytic enzymes that may aid putrefaction. The airlock was tightly covered to facilitate spontaneous fermentation, and the setup was kept at room temperature (28\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C). However, the airlock was loosened after 24 hours to release carbon dioxide. The jar was then stirred to encourage natural microbial growth. The medium was allowed to ferment for 5 days. At day 3, some of the sample was collected and filtered using a 0.4 mm membrane. The sample was refrigerated at -4 \u003csup\u003eo\u003c/sup\u003e C for analysis (Guarner and Schaafsma, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e1998\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Quantification of Proximate Composition\u003c/h2\u003e \u003cp\u003eProximate composition of the fermented samples was determined as described by the Association of Official Analytical Chemists (AOAC, 2012).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Amino acid content (LC-MS)\u003c/h2\u003e \u003cp\u003eThe fermented \u003cem\u003eA. garckeana\u003c/em\u003e were added into water containing 13C, 15N-labeled algal amino acid mix (Cambridge Isotope Laboratories, Inc., USA) at a concentration of 10 g/mL. The amino acids in the diluted \u003cem\u003eA. garckeana\u003c/em\u003e sample were subsequently analyzed in LC-MS/MS Sciex QTRAP 6500\u0026thinsp;+\u0026thinsp;triple-quadrupole-trap MS/MS (Jander et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Vadassery et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Crocoll et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Also, the amino acid was separated with a Zorbax Eclipse XDB-C18 column (50 \u0026times; 4.6 mm, 1.8 \u0026micro;m, Agilent Technologies). The mobile phase consisted of two solvents, A (water with 0.1% formic acid) and B (acetonitrile with 0.1% formic acid). The flow rate was maintained at 1.1 mL per min with the elution profile ran as follows: 0\u0026ndash;1 min, 3% B; 1-3.8 min, 3\u0026ndash;50% B; 3.8\u0026ndash;3.9 min 50\u0026ndash;100% B, 3.9-5 min, 100%B; 5-5.1 min, 100-3% B, 5.1-7 min, 100% B. The column was held at a constant temperature (27\u0026deg;C). The analyte parent ion and product ion were monitored closely using the mass spectrometer in positive ionization mode (MRM modus) for detection Jander et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). Both Q1 and Q3 quadrupoles were maintained at unit resolution. Furthermore, Analyst 1.5 software (Sciex) was used for data acquisition and processing. Using the method described by Muni et al. (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), individual amino acids in the sample were quantified with 13C and 15N-labeled amino acids, respectively.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5 GC-MS Analysis of Vitamins\u003c/h2\u003e \u003cp\u003eThe samples (AGW and AGS) were analyzed using GC-MS to identify and quantify the vitamins present. The analysis was performed with an Agilent Technologies 7890 GC system coupled to a 5975 Mass Spectrometry detector. Separation was achieved using an Agilent HP-5MS capillary column (30 m \u0026times; 0.320 mm \u0026times; 0.25 \u0026micro;m film thickness). Helium served as the carrier gas at a constant flow rate of 0.5 mL/min, and an injection volume of 1 \u0026micro;L was introduced into the system.\u003c/p\u003e \u003cp\u003eThe oven temperature was initially set at 80\u0026deg;C for 2 minutes, then increased at 10\u0026deg;C per minute to a final temperature of 240\u0026deg;C for 6 minutes. The total run time was 90 minutes. Electron impact ionization was conducted at 70 eV. Vitamins identification and quantification were based on Total Ion Chromatograms (TIC). The resulting spectra were compared with reference spectra in the National Institute of Standards and Technology (NIST) library (Gaithersburg, Maryland, USA) for accurate compound identification (Momodu et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e2.6 Mineral Content Analysis (XRF)\u003c/h2\u003e \u003cp\u003eElemental minerals of the fermented samples were determined by EDXRF analysis using an ARL (Yagi et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). The Quant\u0026rsquo;X system is equipped with a cellulose X-ray filter and a Rh-anode X-ray tube. The system operated within a voltage range of 8\u0026ndash;12 kV and a current intensity of 0.32\u0026ndash;0.34 mA. A Si (Li) detector cooled with liquid nitrogen was used to detect the emitted X-rays. Each sample was analyzed for 120 seconds, with an average dead time of approximately 50%. Data processing was performed using WINTRACE software version 4.1, build 9. All analyses were conducted in triplicate for each plant sample to ensure accuracy and reproducibility.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e2.7 Data Analysis\u003c/h2\u003e \u003cp\u003eThe results obtained in triplicate were expressed as mean values and standard deviation (SD). All analyses were performed using Microsoft Excel software\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results and Discussion","content":"\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e3.1 Proximate Composition of A. garckeana\u003c/h2\u003e \u003cp\u003eThis study revealed the highest moisture and carbohydrate contents in AGS at day 5, but the lowest in AGW at day 3 (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Protein and Ash content reduced with increasing fermentation duration for both samples. Lipid content was highest in AGW (5 days) but lowest in AGS (5 days). The increased moisture observed in the current study may indicate accelerated fermentation due to the growth of desirable microorganisms and the influence of their metabolites, such as acids and alcohols, as reported in similar fruit fermentations (Di Cagno et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Driven by microbial enzymes, the increased carbohydrate in this study may indicate the presence of resistant starches. During fermentation, starches are converted into simpler and digestible forms like acetic acid and lactic acid. Acetic acid and lactic acid lower pH, promote the shelf life of the sample, and enhance the digestibility of foods, positioning fermented \u003cem\u003eA. garckeana\u003c/em\u003e as a valuable energy source for populations at risk of protein\u0026ndash;energy malnutrition (Nkafamiya et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Hence, supports SDG 2 on Zero Hunger. The decrease in protein content with time corroborates earlier findings by Falana et al. (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), who explained that fermenting microorganisms may have broken down the protein into amino acids and peptides. The increased lipid content may mean microbial synthesis and accumulation of lipid metabolites, consistent with observations by Falana et al. (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eProximate Composition of the Two Fermented Varieties of \u003cem\u003eA. garckeana\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eParameters\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAGW (3 Days)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAGW (5 Days)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAGS (3 Days)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAGS (5 Days)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMoisture\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e60.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.005\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e54.54\u0026thinsp;\u0026plusmn;\u0026thinsp;0.010\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e59.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.010\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e57.83\u0026thinsp;\u0026plusmn;\u0026thinsp;0.040\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eProtein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e3.44\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0004\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e2.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e5.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0005\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e2.42\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0015\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLipid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e5.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.005\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e6.44\u0026thinsp;\u0026plusmn;\u0026thinsp;0.000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e3.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.005\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e2.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.000\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCrude fiber\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e25.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.005\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e23.57\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0005\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e21.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e22.87\u0026thinsp;\u0026plusmn;\u0026thinsp;0.000\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAsh content\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e5.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e4.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e3.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.004\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e3.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCarbohydrate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.79\u0026thinsp;\u0026plusmn;\u0026thinsp;0.000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e8.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0005\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e7.36\u0026thinsp;\u0026plusmn;\u0026thinsp;0.004\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e14.15\u0026thinsp;\u0026plusmn;\u0026thinsp;0.004\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003e\u003cb\u003eKey: AGW\u003c/b\u003e: \u003cem\u003eA. garckeana\u003c/em\u003e fermented in water, \u003cb\u003eAGS\u003c/b\u003e: \u003cem\u003eA. garckeana\u003c/em\u003e fermented in sugarcane juice\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e3.2 LC-MS Mineral Composition of A. garckeana\u003c/h2\u003e \u003cp\u003eSeventeen amino acids were identified in samples (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), with glutamic acid being the most abundant and tyrosine the least in both fermented samples. Such amino acids play crucial physiological roles in energy metabolism, neurotransmission, tissue repair, and collagen synthesis, suggesting that fermented \u003cem\u003eA. garckeana\u003c/em\u003e may have cognitive and physiological health-boosting properties. This is consistent with previous compositional studies on \u003cem\u003eA. garckeana\u003c/em\u003e (Nkafamiya et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Bioltif et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). The sugar content of sugarcane juice may be bio-enhancing, leading to the optimized nutrient conversion and synthesis of essential compounds in the sample (Di Cagno et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). The increased proline and leucine with fermentation time indicate extensive proteolysis and amino acid liberation, enhanced digestibility, and metabolic utilization (Nkafamiya et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e3.3 GC-MS Vitamins Composition of A. garckeana\u003c/h2\u003e \u003cp\u003eNotably, vitamins A, B9, and C increased significantly in sugarcane-fermented samples after five days (Figs.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e\u0026ndash;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). This reinforces the role of sugar-rich substrates in microbial synthesis of bioactive compounds (Falana et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Higher mineral concentrations, including calcium, potassium, and chlorine, were recorded in sugarcane-fermented samples, all of which are critical for muscle function, nerve transmission, and electrolyte regulation (Ochokwu et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). In line with our study, Sulieman (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) asserted that fermentation improves the nutritional and sensory properties of local fruits.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003e3.4 Mineral Composition of A. garckeana by X-ray fluorescence\u003c/h2\u003e \u003cp\u003eSeven macro minerals (Oxygen, magnesium, phosphorus, chlorine, potassium, calcium, and sulfur), six microminerals (Copper, zinc, chromium, manganese, iron, and vanadium), and two other biological essential elements (silicon and silver) were identified in the fermented samples (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The elevated potassium and calcium concentrations observed in sugarcane-fermented samples align with prior studies by Ochokwu et al. (\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) and Momodu et al. (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), which highlighted the mineral richness of \u003cem\u003eA. garckeana\u003c/em\u003e. The presence of biologically useful trace elements such as iron and zinc may be linked to hemoglobin synthesis, enzymatic regulation, and immune defense (Abass and Ahmed, \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Also, trace elements silicon and vanadium, which are associated with bone metabolism and catalytic enzyme functions (Yagi et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2013\u003c/span\u003e), emphasize the potential of the fermented samples as functional foods for well-being and support SDG 3 on Good Health and Well-being.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eComparative Elemental Mineral Content of the Two Fermented Varieties of \u003cem\u003eA. garckeana\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eS/n\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eElement\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSymbol\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAFW 3 Days\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAFW 5 Days\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAFS 3 days\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eAFS 5 Days\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOxygen\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eO\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e12.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e21.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e25.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMagnesium\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCopper\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCu\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.8\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eZinc\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eZn\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSilicon\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSi\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e7.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e5.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePhosphorus\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eP\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.4\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eChlorine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e52\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePotassium\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eK\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e8.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCalcium\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e5.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eVanadium\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eV\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eChromium\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCr\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eManganese\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMn\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIron\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eFe\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSulfur\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSilver\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003e\u003cb\u003eKey: AGW\u003c/b\u003e: \u003cem\u003eA. garckeana\u003c/em\u003e fermented in water, \u003cb\u003eAGS\u003c/b\u003e: \u003cem\u003eA. garckeana\u003c/em\u003e fermented in sugarcane juice\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"4. Conclusion","content":"\u003cp\u003eThis study demonstrates that medium and duration of fermentation influence the nutritional composition of \u003cem\u003eAzanza garckeana\u003c/em\u003e. Sugarcane juice and extended duration optimized the levels of vitamins (A, B9, and C), carbohydrates, and lipids while promoting the release of amino acids such as glutamic acid, leucine, and proline, which contribute to improved digestibility and physiological performance. Vital macro- and microelements\u0026mdash;particularly potassium, calcium, zinc, and iron, which support metabolic, immune, and hematological health, were present.\u003c/p\u003e \u003cp\u003eThese results affirm that fermentation not only preserves \u003cem\u003eA. garckeana\u003c/em\u003e but also transforms it into a functional food with enhanced nutritional and health-promoting potential. Thus, beyond the nutritional relevance of fermentation, it serves as a bio-enhancing and sustainable process that promotes microbial vitamins and mineral synthesis, and contributes to the realization of key Sustainable Development Goals (SDGs), especially SDG 2 (Zero Hunger), SDG 3 (Good Health and Well-being), and SDG 12 (Responsible Consumption and Production).\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eData Availability Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data generated or analysed during this study are included in this published article [and its supplementary information files].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThere are no competing interests. All authors certify that they have no affiliations with or involvement in any organization or entity with any financial interest or non-financial interest in the subject matter or materials discussed in this manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors have not received any funding for this study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors' contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMBF developed the review protocol and study design. MBF and HT drafted the manuscript; MRB and MAD were responsible for data analysis and interpretation. MBF and QON edited the manuscript and contributed to data analysis and interpretation. All authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClinical trial\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics, Consent to Participate, and Consent to Publish declarations\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe fruits of \u003cem\u003eAzanza garckeana\u003c/em\u003e (F. Hoffm.) Exell \u0026amp; Hillc., used in this study, were procured with local guidelines from local herb sellers.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePermission to Collect Plant Specimen\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAppropriate permission was received for the procurement of the plant part used in this study.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAbass AA, Ahmed GA. 2017. 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J Biol Agric Healthc. 2015;5(15):26\u0026ndash;31.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOsum FI, Okonkwo TM, Okafor GI. Effect of processing methods on the chemical composition of \u003cem\u003eVitex doniana\u003c/em\u003e leaf and leaf products. Food Sci Nutr. 2013;1(3):241\u0026ndash;5.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSulieman AMEH. \u003cem\u003eAzanza garckeana\u003c/em\u003e: Distribution, composition, nutritive value and utilization. Wild Fruits: Composition, Nutritional Value and Products. Cham: Springer Int Publ; 2019. pp. 379\u0026ndash;93.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVadassery J, Reichelt M, Jimenez-Aleman GH, Boland W, Mith\u0026ouml;fer A. Neomycin inhibition of (+)-7-iso-jasmonoyl-L-isoleucine accumulation and signaling. J Chem Ecol. 2014;40(7):676\u0026ndash;86.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYagi S, Rahman AE, Elhassan GO, Mohammed AM. Elemental analysis of ten Sudanese medicinal plants using X-ray fluorescence. Appl Res Ind Eng. 2013;1(1):49\u0026ndash;53.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYusuf AA, Garba R, Alawode RA, Adesina AD, Oluwajobi I, Ariyeloye SD, Berinyuy BB. Effect of drying methods and extractants on secondary metabolite compositions of \u003cem\u003eAzanza garckeana\u003c/em\u003e pulp and shaft. Noble Int J Agric Food Technol. 2020;2(1):1\u0026ndash;7.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"discover-food","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"discoverfood","sideBox":"Learn more about [Discover Food](https://www.springer.com/44187)","snPcode":"","submissionUrl":"","title":"Discover Food","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Azanza garckeana, Fermentation, Nutritional Composition, Amino acids, Sustainable Development Goals","lastPublishedDoi":"10.21203/rs.3.rs-8627658/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8627658/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eTraditionally consumed as food and medicine, the impact of fruit fermentation of \u003cem\u003eAzanza garckeana\u003c/em\u003e (F. Hoffm.) Exell \u0026amp; Hillc. in distilled water (AGW) and sugarcane juice (AGS) for 3 and 5 days, and the nutritional composition was examined. The moisture level of AGW and AGS decreased (60.05% and 59.04% to 54.54% and 57.83%). Protein reduced from 3.44% (AGW) and 5.81% (AGS) to 2.81% and 2.42%, respectively. Lipid increased from 5.22 (AGW) to 6.44%, but decreased from 3.11 (AGS) to 2.72%. Crude fiber slightly reduced (25.34\u0026ndash;23.57%) in AGW and increased (21.35\u0026ndash;22.87%) in AGS. Carbohydrate increased from 0.79\u0026ndash;8.33% to 7.36\u0026ndash;14.15% in AGW and AGS, respectively. Liquid Chromatography-Mass Spectrometry (LC-MS) identified 16 amino acids; glutamic acid, arginine, and cysteine were abundant, while tyrosine was sparse. Gas Chromatography-Mass Spectrometry (GC-MS) identified higher vitamins C, B9, and A on day 5 and in AGS. X-ray fluorescence (XRF) revealed macro-minerals (O, Mg, P, Cl, K, Ca, S), micro-minerals (Cu, Zn, Cr, Mn, Fe, V), and trace elements (Si, Ag). Cl and O decreased (60\u0026ndash;42 and 12.1\u0026ndash;21) in AGW, but increased (39\u0026ndash;52) in AGS. Fermented \u003cem\u003eA. garckeana\u003c/em\u003e is an affordable, nutritious, and sustainable bioresource that addresses zero hunger, poverty alleviation, good health, and economic empowerment.\u003c/p\u003e","manuscriptTitle":"Influence of Fermentation in Water and Sugarcane Juice on Nutritional Composition of Azanza garckeana Fruit","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-13 17:18:03","doi":"10.21203/rs.3.rs-8627658/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"editorInvitedReview","content":"","date":"2026-03-26T14:54:55+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"219514358585050468099229137643898931566","date":"2026-03-17T14:01:41+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-03-08T15:09:09+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"51767882650823051682325807315970567095","date":"2026-03-03T08:02:06+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"113501599976490499357748252340418990705","date":"2026-02-23T11:57:11+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-02-10T11:32:16+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2026-02-06T11:22:40+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-02-04T12:39:57+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-02-03T13:13:02+00:00","index":"","fulltext":""},{"type":"submitted","content":"Discover Food","date":"2026-02-03T12:29:03+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"discover-food","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"discoverfood","sideBox":"Learn more about [Discover Food](https://www.springer.com/44187)","snPcode":"","submissionUrl":"","title":"Discover Food","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"c9f1d0ef-4c98-433e-8d2c-74be698cb6bd","owner":[],"postedDate":"February 13th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-02-13T17:18:03+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-13 17:18:03","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8627658","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8627658","identity":"rs-8627658","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}