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. 2025 Oct 1;15:34129. doi: 10.1038/s41598-025-17960-9

Fortification of non-dairy milk with date fruit, mustard seed, and turmeric: nutritional quality, probiotics viability, antimicrobial and antioxidant potentials

Clement Olusola Ogidi 1,✉, Oluwatoyin Ajoke Oladeji 1, Kayode Olayele Karigidi 2, Olugbenga Oludayo Oluwasina 3, Foluso Olutope Adetuyi 2
PMCID: PMC12489101  PMID: 41034312

Abstract

Awareness about medicinal values of plant-based milk as functional foods has created a need to switch from chemically fortified food products to naturally healthy plant-based foods, which offer essential nutrients required for various metabolic activities, and physiological stability. Therefore, this study was designed to enhance the nutrient contents, consumer acceptability, probiotics viability in plant-based milk from coconut, soybean, tiger nut fortified with extracts from Phoenix dactylifera (date fruit), Brassica juncea (mustard seeds), and Curcuma longa (turmeric). The bioactive compounds in extracts from date fruit, mustard seed, and turmeric were identified using Gas Chromatography/Mass Spectrometry (GC-MS). Nutrient contents, organoleptic property, microbiological quality, probiotics viability, inhibition of diarrheagenic Escherichia coli, and antioxidant potentials of the fortified plant-based milk were determined using standard methods. Some of the bioactive compounds identified in plant extracts with the aid of GC-MS were isoquercetin, cis-oleic, sinigrin, erucic acid, α-phellandrene, zingiberene, and α – curcumene. The highest protein content (5.10%) was obtained in soy milk fortified with mustard seed (97%SM + 3%MS). Coconut milk (100%CM) had the lowest crude fibre of 0.30%. Plant milk (97%CM + 3%DG) supported the growth (× 105 CFU/mL) of Lactobacillus fermentum, Lactobacillus acidophilus, and Lactobacillus pentosus with values of 6.20, 5.80, and 4.60, respectively. Plant milk (97%TM + 3%MS) had the highest scavenging activity (89.10%) against 1,1-diphenyl-2-picrylhydrazyl (DPPH), while (97%SM+3%MS) showed the highest activity (305.10 µmol TE/mL) against ferric reducing antioxidant power (FRAP). Plant-based milk fortified with extracts from date fruit, mustard seed, and turmeric displayed nutraceutical potentials and could be considered as promising alternative source of functional foods.

Supplementary Information

The online version contains supplementary material available at 10.1038/s41598-025-17960-9.

Keywords: Functional foods, Nutrient, Secondary metabolites, Phytomedicine, Diarrhea, Consumer acceptability

Subject terms: Antimicrobials, Applied microbiology, Industrial microbiology

Introduction

Over the past decade, research emphasis on the food product development is ongoing to address the challenges of food insecurity as well as malnutrition by producing newer healthy functional foods as alternative-medicinal foods to meet the demand of consumers1. Plant-based milk from cereals namely; oat, rice, corn, spelt milk, legumes such as soybean, peanut, lupin, cowpea, nut group like coconut, hazelnut, almond, walnut, seeds (sesame, flax, hemp, sunflower), pseudo-cereal (quinoa, teff, amaranth), fruits, and vegetables are alternative options desired by many individuals for a healthy lifestyle and thus, serve as substitute to foods from animal milk products2–4. Plant-based milk substitutes is indispensable in the vegan food industry, serving both as key ingredients in a variety of food products and as a main alternative source of essential nutrients to human. The bio-functional properties of plant-based milk address diverse needs and lifestyles by enhancing energy, combating aging, fatigue, stress, and targeting specific health conditions5,6. The medicinal values of plant milk have expanded its uses as a viable alternative to dairy products, particularly for managing health issues such as allergies, lactose intolerance, calorie concerns, and hypercholesterolemia associated with consumption of cow milk7. Dairy milk is often more expensive due to higher production cost, storage challenges, and limited accessibility, especially for populations in low- and middle-income countries, compared to plant-based milk that is now being perceived as a direct equivalent or even more healthful alternatives to specific consumers8.

Plant-based milk serves as an alternative for individual seeking affordable and accessible nutritional sources to combat malnutrition, and any symptoms of nutrient deficiencies. Plant-based milk are sustainable diet that are culinarily accepted, and can be produced in an ecofriendly manner without pollution, acidification, eutrophication or any adverse effect on environment witnessed during the rearing of livestock for milk production9. Non-dairy milk from coconut contains medium-chain triglycerides fatty acids together with vitamin E, which support digestion, act as anti-carcinogenic, anti-microbial, and slow down the aging process as antioxidants drink without allergenic reactions10. Milk from soybean attracts substantial number of consumers, which therefore, emerged as a prominent vegetable milk of high-quality due to its nutrient contents, absence of cholesterol, and with abundant heart-healthy polyunsaturated fatty acids11. Likewise, milk from tiger nut is often consumed by many people for health benefits to prevent cardiac arrest, constipation, and diarrhea due to the presence of oleic acid, vitamins, minerals, and antioxidant compounds12. However, nutritional quality of plant-based milk still remained unbalance, and their sensorial properties like mouth feeling, flavour, and aroma are less accepted when compared to animal milk13,14. These drawbacks can be mitigated by typically supplementing the plant milk with treasurable secondary metabolites from edible plants to enhance their nutritive values, improve consumer acceptability, and shelf-life stability.

Edible plants like P. dactylifera fruit offers a wide array of health benefits as antioxidant to prevent cancer, neurodegenerative conditions (Alzheimer’s and Parkinson’s diseases), and cardiovascular problem15. Date fruits of different cultivars exhibit antimicrobial activities against microorganisms like Staphylococcus aureus, Escherichia coli, Bacillus cereus, Shigella sonnei, S. typhi, Serratia marcescens, Aspergillus flavus, Penicillium chrysogenum, and Aspergillus niger16,17. B. juncea (mustard seed) is recognized for its nutritional value and traditional uses as a food spice. In folk medicine, it is used as an appetite stimulant, an expectorant, and as a remedy for various health conditions such as gastrointestinal issues, skin diseases, arthritis, foot ache, lumbago, and rheumatism18. C. longa (turmeric) is an excellent natural anti-inflammatory, commonly use to aid digestion, to improve intestinal flora, and widely accepted due to its peppery, slightly bitter flavour, and a mild fragrance19. Hence, the nutrients, functional properties, and consumers’ acceptability of plant-based milk supplemented with extracts from medicinal plants has not been adequately reported. This research study was therefore, designed to reveal the nutrient contents, organoleptic property, viability of Lactic Acid Bacteria (LAB), inhibition of diarrheagenic E. coli, and antioxidant potentials of the formulated non-dairy milk enriched with extracts from selected edible plants namely; date fruits, mustard seed, and turmeric rhizome.

Materials and methods

Reagents used

Gallic acid, Folin-Ciocalteu, 2,4,6-tri(2-pyridyl)-s-triazine, and sodium nitrate were obtained from Sigma-Aldrich (St. Louis, MO, USA). Sodium acetate, ferric chloride, and aluminum chloride were produced by Poole chemical (Dorset, England). Sodium hydroxide, hydrochloric acid, potassium persulfate, 1,1-diphenyl-2-picrylhydrazyl (DPPH), ethanol, trichloroacetic acid were from Merck (Darmstadt, Germany). Trolox, iron (II) sulphate, butylated hydroxytoluene (BHT) were products of Sigma-Aldrich (Steinheim, Germany). Nutrient agar (NA), potato dextrose agar (PDA), De Man Rogosa and Sharpe (MRS) agar were from HiMedia (Maharashtra, India). Sorbitol MacConkey Agar, Cefixime and Tellurite MacConkey Agar, and tryptone soy broth were from Oxoid (Basingstoke Hampshire, UK).

Collection of samples

Tiger nut, soybeans, turmeric rhizome, and coconut were purchased from vendors in Okitipupa market, Ondo state. Mustard seed was purchased from fruit retailer shop in Lagos state. Diarrhoeagenic strains of E. coli, and E. coli O157:H7 were collected from Nigerian Institute of Medical Research (NIMR), Lagos.

Preparation of extracts from date fruit, mustard seed and turmeric

Date palm fruit flesh seed was manually removed, and the flesh part (100 g) was blended with water (400 mL) to obtain a fine paste. Dried mustard seed (100 g) was pulverized and mixed with 400 mL of warm water. This solution was properly mixed together before sieving to remove undissolved particle. Turmeric powder (100 g) was mixed with warm water (400 mL) and then filtered to separate the solid content. All the extracts were filtered through sterile muslin cloth.

GC-MS analysis of extract from date, mustard seed and turmeric

Gas Chromatography/Mass Spectrometry (GC-MS) analysis of each plant extract was performed using a multidimensional gas chromatography coupled with gas chromatography-mass spectrophotometer (Shimadzu Japan) equipped with non-polar and polar double capillary columns (25.0 m×0.25 μm i.d., 0.25 μm df). High purity helium was used as the carrier gas at a constant flow rate of 0.99mL/min. Sample (1 µL) was injected (split ratio 100:1) into GC and GCMS using AOC-2Oi; auto injector for analysis. The initial temperature was set at 60 °C, heated at a rate of 3 °C/min to 280 °C and held isothermally for 6 min. Ion source temperature was set to 200 °C, while the interface was set at 250 °C, solvent cut time was 3 min. Electron impact (EI) ionization mode was 70ev, and the linear velocity of the column was 36.8 cm/sec. The identification of the various components was based on comparison of their mass spectra with those of NIST Library mass Spectra data base and mass spectra from Literature.

Preparation of milk from coconut

Coconut milk was prepared according to the method described by Tulashie et al.10 with slight modifications. Coconuts were de-husked, and a white fleshy solid endosperm was obtained. The white fleshy part (500 g) was blended with water (1 L) and boiled at 50 °C for 3 min. The viscous slurry resulting from the mixture was filtered using a muslin cloth and pasteurized at 70 °C for 10 min.

Preparation of milk from soybean

Dry soybean (500 g) was washed and soaked in 2 L of water for 12 h for ease dehulling. It was blanched at 60 °C for 30 min to remove the bitterness, anti-nutritional factors, and to improve nutritional values20. The soybean was blended with water (1:3 w/v) using a blender. The resultant slurry was filtered using a sterile muslin cloth. The filtrate was boiled at 70 °C for 10 min and allow it to cool.

Preparation of milk from tiger nut

Milk from Tiger nut was produced by using the method of Adebayo-Oyetoro et al.21 with little modification. The tiger nuts were sorted to remove poor-quality nuts that could impair the milk’s flavour. Tiger nuts (700 g) were rinsed with water, soaked in water (2 L) for 24 h, wet milled with 2 L of water in a blender. This was followed by filtration using a muslin cloth to remove the chaff from the milk and pasteurized at 70 °C for 10 min.

Fortification of plant-based milk with extract from date fruit, mustard seed, and turmeric

To the various concentrations (95–99%) of plant-based milk from coconut, soybean, and tiger nut, different percentage (1–5%) of extracts from date fruit, mustard seed, and turmeric were added. The formulation containing 97% plant milk and 3% extract (v/v) was the most widely accepted by panelists. Subsequently, further studies focused on these particular samples, which were stored in a refrigerator at 4 °C.

Proximate and mineral analysis of plant-based milk

The moisture, ash, crude fiber, fat, crude proteins and carbohydrates contents of plant-based milk were determined by using methods of Association of Official Analytical Chemists22. Atomic Absorption Spectrophotometer (Buck Scientific, Model 200 A/200) was used to determine Zn, Fe, Ca, P, while a flame photometer (Jenway PFP 7, Staffordshire, UK) was used to determine Na, and K contents in the plant milk. Each of the metal standard solution at 1000 µg/mL in 1% v: v HNO3 was used for calibration of atomic absorption spectroscopy.

Organoleptic properties of plant milk supplemented with plant extracts

The sensory assessment was conducted after the approval of the Research committee on sensory evaluation and consumer protection (Ref: OAUSTECH/FST/2024-007). Organoleptic properties; colour, aroma, taste, and flavour were carried out using 15-member semi-trained panelists on a 9-point hedonic scales with (9) = extremely like and (1) = extremely dislike. The samples were presented to the panelists using white glass cups.

Assessment of microbiological quality of plant- based milk

For the microbiological quality, isolation of bacteria and fungi were carried out on nutrient agar (NA) and potato dextrose agar (PDA), respectively. All media were prepared in accordance with the instructions provided by the manufacturer, and sterilized at 121 °C for 15 min. Dilution factors of (102 to 104) was pour plated on sterile molten agar, stirred, and allowed to set. The plates were incubated at 37 °C for 24 h for bacteria, and 30 °C 48 h for fungi. Colonies on each plate were enumerated in colony forming unit per milliliter (CFU/mL) after incubation.

Viability of lactic acid bacteria and inhibition of diarrheagenic E. coli

Lactobacillus acidophilus, Lactobacillus fermentum and Lactobacillus pentosus were isolated from Kunu-zaki, a popular non-alcoholic fermented beverage from millet, sorghum, or maize. The identity of lactic acid bacteria was confirmed with Bergey’s manual of determinative bacteriology23, and probiotic characteristics were determined using the methods of Jose et al.24. To prepare inoculum cultures, sterilized (at 121 °C for 15 min) de Man, Rogosa and Sharpe (MRS) broth (10.0 mL) in different test tubes were inoculated with 100 µL of each strain, followed by incubation at 37 °C for 48 h. Lactic acid bacteria inoculum culture (2% v/v; containing 105 colony forming units (CFU) /mL of individual strains) was added to each plant milk (100 mL) individually, and incubated at 40 °C for 12 h. the sample without lactic acid bacteria serve as control. After incubation, growth rate of probiotics; Lactobacillus acidophilus, Lactobacillus fermentum and Lactobacillus pentosus was quantified by serially diluting (100 µL) in sterile distilled water up to 10− 4, and 100 µL was placed on MRS agar plates using pour plate method. Petri dishes were incubated at 37 °C for 48 h. Thereafter, colonies of Lactobacillus acidophilus, Lactobacillus fermentum and Lactobacillus pentosus were counted, and reported as colony forming unit per milliliter (CFU/mL).

For the inhibition of diarrheagenic E. coli strains by plant-based milk, method of Kivanc and Yapici25 was adopted with slight modifications. Briefly, pure colony of diarrheagenic E. coli were transferred into sterile tryptone soy broth, and incubated for 24 h at 37 °C to obtain activated inoculum. Plant milk (9.0 mL) with and without plant extract were separately transferred into a sterile bottle, and 1.0 mL of diarrheagenic E. coli strains (1.0 × 106 CFU/mL) conforming to McFarland standard was added to the test tubes. Each test tube was incubated at 37 °C for 24 h. Thereafter, 0.1 mL from dilution × 103 of each sample was transferred to Petri dish to estimate the growth of E. coli and E. coli O157:H7 on sorbitol MacConkey Agar, Cefixime and Tellurite MacConkey Agar, respectively. Uninoculated plant-based milk served as a control.

Determination of total phenolic and flavonoid contents

Extracts were obtained from the plant milk by taken 10 mL of each plant milk into 2.5 mL of distilled water. The mixtures were centrifuged at × 20,000 g for 10 min, and the supernatant was utilized to determine the total phenolic and flavonoid contents. Total phenolic content was quantified using Folin-Ciocalteu reagent26. Total phenolic content in each extract was measured at 725 nm, expressed as milligram gallic acid equivalent per gram of sample (mg GAE/mL) using the standard curve of gallic acid, R 2 = 0.9985. The flavonoid content was quantified using aluminum chloride assay with absorbance measured at 510 nm27. The total flavonoid content of each sample was calculated as milligram quercetin equivalent per gram of sample (mg QE/mL) using standard curve of quercetin, R 2 = 0.9975.

Antioxidant activities of plant-based milk

Assay method described by Xiao et al.28 was employed for DPPH radical scavenging at 517 nm. DPPH radical scavenging activity was calculated as DPPH scavenging activity (%). Ferric-reducing antioxidant power assay and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging activity was determined using the methods of Xiao et al.28. The results for FRAP and ABTS were expressed as µmol of trolox equivalent per milliliter (µmol TE/ mL).

Statistical analysis

Each experiment was performed in replicates (n = 3). Statistical analysis was conducted on data obtained using one-way Analysis of Variance (ANOVA) with SPSS Statistics version 22 for Windows (SPSS Inc., Chicago, IL, USA). The differences between means were considered using the New Duncan Multiple Range Test at a significance level of P ≤ 0.05. Pearson’s correlation analysis was used to assess relationships between bacterial responses in the presence of plant-based milk. A correlation heatmap was generated to visualize the strength and direction of linear associations the corrplot package in R (https://github.com/taiyun/corrplot) in R version 4.3.1 (https://www.r-project.org/).

Results and discussion

GC-MS profile of bioactive compounds in extract of date fruit, mustard seed and turmeric

Edible plants contain several bioactive compounds that can be utilized for new product development. Figures S1 to S3 show the chromatograms of bioactive compounds, while Tables S1 to S3 show the peak areas, and identified bioactive compounds in extract of date fruit, mustard seed, and turmeric. The bioactive compounds in date fruit extract are isoquercetin, linolenic acid, β –D-glucopyranose, linoleic acid, cis-oleic, linolenic acid, and others. Findings of Maqsood et al.29 revealed the presence of gallic acid, p-coumaric acid, quercetin, carotenoids, α-tocopherol acetate, ergasterol, estrone, brassicasterol, and tannins in date fruit. These compounds contribute to the notable antioxidant potential, antimicrobial property, anticancer activity, and antidiabetic effects of date fruit16,30. These therapeutic attributes underscore its nutraceutical relevance and suitability for diverse food formulations. GCMS revealed the presence of elaidic acid, gluconapin, arachidic acid, glucoerucin, sinigrin, and erucic acid in the mustard seed extract (Table S2). Findings of Khan et al.31 revealed higher proportion of gluconapin, 3-,5′-dimethoxyacetophenone, oleic acid, 4-methylpentyl S-2-(dimethylamino) ethyl propylphosphonothiolate, and erucic acid in mustard seed. The bioactive compounds in mustard seed contributed to plethora of pharmacological activities such as antioxidant, anti-inflammatory, antimicrobial and antidiabetic effects32.

The bioactive ingredients detected in turmeric using GC-MS are α-phellandrene, zingiberene, α–curcumene, β– sesquiphellandrene, Delta (sup 9) -cis-oleic acid, farnesene,  and others. Earlier findings of Ogidi et al.53 reported the presence of α -pinene, camphene, β -myrcene, α-phellandrene, α-curcumene, linalol, zingiberene, farnesene, β -citronellol, and β-sesquiphellandrene in turmeric essential oil. The functional activities of bioactive compounds in turmeric promote individual health, and have sparked growing global interest in nutraceutical as a complement to pharmaceuticals33. The bioactive compounds in edible plants have demonstrated biological potentials. Therefore, combining two or more edible, and medicinal plants from different species such as herbs, fruits, vegetables, seeds, and other plant parts can produce next-generation nutritionally fortified foods. This conforms to the findings of McClements34 who revealed the advantage of supplementing plant-based milk with other nutritious sources as an ideal vehicle for introducing essential nutraceuticals into the human diet. These innovative food formulations have the potential to effectively address the widespread issue of low macronutrient, and micronutrient density in plant-based milk consumed by humans.

Nutritional composition of plant-based milk fortified with extract from date fruit, mustard seed, and turmeric

Table 1 shows the proximate composition of plant-milk containing extracts of date, mustard seed and turmeric. The moisture content of studied plant milk ranged from 78.20 to 92.40%. High moisture content in food promotes enzymatic activity and increase susceptibility to microbial growth, indicating a shorter shelf life and classifying the food as highly perishable35. Microorganisms thrive in fruit juices, and plant milk by utilizing the nutrients such as simple sugars, amino acids, and mineral available in these fluids to support their growth and metabolism36. Therefore, plant-based milk should be stored at very low temperature to minimize the growth of spoilage microorganisms, despite the addition of plant extracts or any natural preservative. Coconut milk has the lowest (P ≤ 0.05) crude fibre of 0.30% but increased (P ≤ 0.05) to 0.90% when 3% of date fruit was added to the 97% coconut milk (97%CM + 3%DG). The addition of date fruit contributed to the crude fiber content of milk from soybean (97%SM + 3%DG), and tiger nut (97%TM + 3%DG) as 2.10%, and 2.70%, respectively. Date fruit is an excellent source of both insoluble, and soluble dietary fiber, which has demonstrated beneficial physiological effects on human health by helping to prevent cardiovascular disease, diverticulosis, constipation, irritable bowel syndrome, and lowering blood glucose37,38. The plant milk, both with and without extract from date fruit, mustard seed, and turmeric contain very low-fat content ranging from 0.60 to 1.70%. Plant-based milk is gaining significant attention due to its low-fat content, and absence of lactose, which contribute to its functional properties, enhanced nutritional value, and establish it as a compelling alternative to animal milk and its products11. Addition of mustard seed increase the protein content of coconut milk from 2.50 to 3.30%, 3.20% to 4.00% in tiger nut milk, and from 4.30% to 5.10% in soy milk. Mustard seeds contain cruciferin and napin as their storage proteins, which when incorporated into food can improve the overall protein quality. The bioactive components, and functional properties of mustard seed further support its use as a valuable source of protein in food formulations, making it a compelling ingredient for the human food industry39.

Table 1.

Proximate composition (%) of plant milk fortified with extract from date fruit, mustard seeds, and turmeric.

Samples Moisture Fat Ash Crude fibre Protein CHO
100%CM 92.40 ± 5.30 a 1.70 ± 0.60 a 0.60 ± 0.00 c 0.30 ± 0.00 c 2.50 ± 0.40 b 2.50 ± 0.03 a
97%CM + 3%DG 91.70 ± 6.20 a 1.40 ± 0.00 b 0.90 ± 0.00 b 0.90 ± 0.00 a 2.60 ± 0.20 b 2.50 ± 0.10 a
97%CM + 3%MS 91.90 ± 3.20 a 1.70 ± 0.60 a 1.09 ± 0.01a 0.60 ± 0.01 b 3.30 ± 0.50 a 1.40 ± 0.01 b
97%CM + 3%TR 91.90 ± 6.20 a 1.50 ± 0.10 b 0.70 ± 0.00 c 0.40 ± 0.01 c 2.20 ± 0.50 c 3.30 ± 0.30 a
100%SM 80.50 ± 3.60 a 0.70 ± 0.00 a 0.90 ± 0.00d 1.10 ± 0.00 d 4.30 ± 0.10 b 12.50 ± 1.03 a
97%SM + 3%DG 80.80 ± 4.30 a 0.60 ± 0.00 a 1.50 ± 0.01b 2.10 ± 0.01 a 4.30 ± 0.20b 10.70 ± 0.40 b
97%SM + 3%MS 80.60 ± 2.80 a 0.80 ± 0.00 a 1.75 ± 0.02a 1.70 ± 0.01 b 5.10 ± 0.40a 10.05 ± 1.20 b
97%SM + 3%TR 80.60 ± 5.30 a 0.60 ± 0.00 a 1.10 ± 0.01c 1.40 ± 0.00 c 4.30 ± 0.10 b 12.00 ± 3.00 a
100%TM 79.10 ± 1.05 a 0.90 ± 0.00 a 1.10 ± 0.01d 2.00 ± 0.00c 3.20 ± 0.00 c 13.70 ± 1.30 a
97%TM + 3%DG 78.20 ± 2.50 a 0.70 ± 0.00 b 1.60 ± 0.03b 2.70 ± 0.30 a 3.20 ± 0.10 c 13.60 ± 2.30 a
97%TM + 3%MS 78.50 ± 4.20 a 0.80 ± 0.00 a 1.90 ± 0.00 a 2.50 ± 0.20a 4.00 ± 0.30 a 12.30 ± 1.00
97%TM + 3%TR 78.70 ± 1.15 a 0.60 ± 0.00 b 1.30 ± 0.03 c 2.30 ± 0.10b 3.10 ± 0.00 c 14.00 ± 3.20 a
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CM, coconut milk; SM, soy milk; TM, tiger nut milk; DG, date fruit gel; MS, mustard seed extract; TR, turmeric extract.

Values within the same column sharing the same alphabet are not significantly different (P ≤ 0.05).

Table 2 shows mineral contents of plant-based milk supplemented with extract from date fruit, mustard seeds, and turmeric. Mustard seed > date fruit > turmeric significantly (P ≤ 0.05) contributed to the amount of potassium, iron, zinc, phosphorous, calcium, and sodium in the supplemented plant milk. Mustards are rich source of health-benefiting minerals such iron, calcium, manganese, phosphorous, magnesium, copper, iron, selenium, and zinc18. Date fruit extract contributed to potassium, iron, and calcium contents of the plant milks. Date fruits are particularly rich source of potassium, followed by calcium, and magnesium. The substantial mineral contents in date fruit contributes to their uses, and potential applications in the production of nutritious, and functional snack bars40. Plant-based milk generally have a very low sodium content, and the incorporation of date fruit, mustard seed, and turmeric furthered reduce the Na concentration. The contents of potassium, calcium, iron, zinc, phosphorus, magnesium, and sodium in plant-based milk such as almond, soy, rice, coconut, and plant-based beverages offer a viable substitute for dairy milk, while providing similar health benefits41.

Table 2.

Mineral composition (mg/100 g) of plant milk supplemented with extract from date fruit, mustard seeds, and turmeric.

Samples K Fe Zn P Ca Na
100%CM 471.40 ± 7.60c 70.10 ± 6.10c 83.60 ± 7.20c 21.30 ± 1.80c 22.40 ± 0.90d 20.90 ± 3.06 a
97%CM + 3%DG 562.50 ± 10.40b 90.90 ± 4.00b 104.60 ± 0.00b 35.80 ± 5.00 a 50.10 ± 1.10b 18.80 ± 2.80 a
97%CM + 3%MS 618.20 ± 9.80a 108.40 ± 4.60a 117.50 ± 7.10a 40.50 ± 3.10 a 58.80 ± 2.30a 19.30 ± 1.10 a
97%CM + 3%TR 482.80 ± 6.30c 76.20 ± 7.20c 88.80 ± 4.90c 26.20 ± 4.40bc 38.80 ± 3.00c 16.30 ± 0.40b
100%SM 661.30 ± 10.30c 107.90 ± 8.00c 103.60 ± 5.50d 57.20 ± 2.50c 61.30 ± 2.50c 30.80 ± 2.10 a
97%SM + 3%DG 701.50 ± 18.50b 135.00 ± 9.10b 131.50 ± 4.60b 84.50 ± 3.90a 86.40 ± 4.10 a 26.40 ± 1.60 a
97%SM + 3%MS 765.90 ± 10.40a 179.10 ± 11.00a 153.40 ± 7.50a 97.00 ± 2.50 a 90.05 ± 5.50 a 28.30 ± 1.10 a
97%SM + 3%TR 673.80 ± 17.10c 112.60 ± 7.00c 119.40 ± 5.80 c 71.60 ± 2.20 b 74.60 ± 4.70b 21.80 ± 0.50b
100%TM 732.80 ± 11.10c 128.30 ± 9.40d 166.80 ± 6.20 c 48.40 ± 2.50 b 46.10 ± 1.90 d 26.10 ± 1.60 a
97%TM + 3%DG 805.60 ± 0.00b 166.40 ± 5.60b 194.80 ± 9.40 b 68.80 ± 5.06 a 68.80 ± 3.30 ab 22.30 ± 2.50 b
97%TM + 3%MS 872.70 ± 7.20a 202.10 ± 9.60a 206.30 ± 4.50 a 72.20 ± 3.60 a 77.40 ± 2.70 a 24.50 ± 2.60 a
97%TM + 3%TR 740.30 ± 9.40c 140.80 ± 5.10c 174.90 ± 7.60c 50.70 ± 4.60 b 57.40 ± 2.50 c 19.90 ± 1.70 c
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CM, coconut milk; SM, soy milk; TM, tiger nut milk; DG, date fruit gel; MS, mustard seed extract; TR, turmeric extract.

Values within the same column sharing the same alphabet are not significantly different (P ≤ 0.05).

Sensory attributes of plant-based milk fortified with extract from date fruit, mustard seed, and turmeric

Figure 1 shows the sensory property of plant-based milk supplemented with extract from date fruit, mustard seed, and turmeric. Plant milk with date fruit (97%CM + 3%DG), (97%SM + 3%DG), and (97%TM + 3%DG) were widely accepted by the panelists possibly due to their sweetness. Dates fruit contain a variety of carbohydrates, notably high levels of soluble sugar like fructose, glucose, sucrose, and maltose, which contribute to its sweetness, and enhance the palatability of foods30. The colour of plant milk were accepted as consumer perceives it. The incorporation of date fruit, mustard seed, and turmeric extracts into plant-based milk highlights the significance of natural colourant from the edible plants, enhanced both visual appeal, and potential health benefits. The colour of food products plays a significant role in consumer meal choices42. With the growing trend of replacing artificial food colorants with natural alternatives, plant extract from date fruit, mustard seed, and turmeric offer a safe, and appealing option for natural pigmentation. The addition (3%) of date fruit, mustard seed, and turmeric extract to plant milk didn’t affect the sensory properties of the milk, but instead contributed to the taste and give it pleasant smell in form of flavour. The overall acceptability of plant-based milk fortified with dates, mustard seeds, and turmeric is high, as confirmed by the panelists, indicating that it effectively meets the specified consumption requirements. The findings of Oduro et al.43 examined the impact of ingredient components on consumer sensory appeal using a 9-point hedonic scale and identified innovative flavors, colour, in three-blend plant-based milk. The optimal formulation of plant-based milk through the combination of various plant materials, incorporation of additional ingredients (sweeteners), enrichment with sources of mineral, and vitamins can enhance the nutritional profile, functional properties of plant-based milk and overall sensory appeal44.

Fig. 1.

Fig. 1

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Sensory properties of plant-based milk supplemented with extract from date fruit, mustard seeds, and turmeric. Key: CM: Coconut milk, SM: soy milk, TM: tiger nut milk, DG: date fruit gel, MS: mustard seed extract, TR: turmeric extract.

Microbiological quality, probiotics viability and Inhibition of pathogens by fortified plant-based milk

Table 3 shows the microbiological quality of non-dairy milk supplemented with date fruit, mustard seed and turmeric. At the initial day, the total bacterial count ranged from 1.00 × 103 to 1.40 × 103 CFU/mL, while the fungal count was 0.40 × 102 CFU/mL to 0.70 × 102 CFU/mL. At the day 2, there were reduction in the microbial load of plant milk fortified with extract from date fruit , mustard seed, and turmeric. Figure 2a shows the bacterial responses to the plant-based milk, while Fig. 2b reveals the correlations between their behavior. The growth of lactic acid bacteria in plant milk conformed to the findings of Harper et al.45, and Al-Zahrani and Shori46. The researchers revealed the survivability of probiotics such as L. rhamnosus, L. plantarum, L. acidophilus, and L. casei in milk from coconut, oat, rice, tiger nut, soybean, almond, and others The incorporation of extract from date fruit , mustard seeds, and turmeric into plant milk supported the growth of L. acidiophilus, L. fermentum, and L. pentosus. Date fruits, mustard seeds and turmeric are richer in indigestible carbohydrates, classifying them as potential prebiotics s with abundant in essential nutrients such as amino acids, vitamins, minerals, dietary fiber, phenolic compounds, which can enhance the growth and activity of probiotics, thereby, supporting a healthy gastrointestinal (GIT) microbiota47–49. Thus, plant-based milk fortified with plant extracts presents a promising option for enhancing gastrointestinal health in consumer.

Table 3.

Microbiological quality (CFU/mL) of plant-based milk stored for 4 days.

Samples /day(s) 0 2 4
TBC × 103 Fungi × 102 TBC × 102 Fungi × 102 TBC × 103 Fungi × 103
100%CM 1.20 0.50 3.00 1.20 2.30 1.10
97%CM + 3%DG 1.20 0.50 0.80 0.10 2.00 1.00
97%CM + 3%MS 1.10 0.60 0.40 0.00 1.80 0.80
97%CM + 3%TR 1.30 0.40 0.60 0.10 2.00 1.10
100%SM 1.00 0.40 2.80 1.40 3.10 1.30
97%SM + 3%DG 1.20 0.40 1.00 0.90 1.60 1.30
97%SM + 3%MS 1.00 0.50 0.80 0.00 1.40 0.90
97%SM + 3%TR 1.10 0.60 0.60 0.00 1.40 1.80
100%TM 1.40 0.70 3.50 1.20 4.50 2.20
97%TM + 3%DG 1.30 0.60 1.00 0.00 2.50 1.50
97%TM + 3%MS 1.40 0.50 0.70 0.00 2.00 1.10
97%TM + 3%TR 1.40 0.70 1.00 0.20 2.20 1.20
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Values are mean of replicates (n = 3).

CM, coconut milk; SM, soy milk; TM, tiger nut milk; DG, date fruit gel; MS, mustard seed extract; TR, turmeric extract; TBC, total bacteria count.

Fig. 2.

Fig. 2

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(a) Scatter plot of lactic acid bacteria (× 105 CFU/mL), diarrheagenic E. coli (× 103 CFU/mL), and (b) correlation heatmap of bacterial responses in the presence of plant -based milk with and without extract from date fruit, mustard seed and turmeric. CM: Coconut milk, SM: soy milk, TM: tiger nut milk, DG: date fruit gel, MS: mustard seed extract, TR: turmeric extract.

The correlation heatmap (Fig. 2b) reveals strong positive correlations between plant-based milk samples fortified with date fruit, mustard seed, and turmeric with theinhibition of diarrheagenic E. coli, which suggests that bioactive compounds in plant extracts incorporated into plant milk play a significant role as antimicrobial and preservative agents. Findings of Çağlar et al.50 revealed that the addition of mustard seeds into meatballs suppressed the growth of aerobic mesophilic bacteria, Enterobacteriaceae, psychrophilic bacteria count, and yeast. Studies of Barakat and Alfheeaid40 attribute the antimicrobial potentials of date fruits against microorganisms to the presence of phytochemicals. Mustard seeds contain bioactive compounds such as allyl isothiocyanate, para-hydroxybenzyl isothiocyanate, glucosinolates (e.g., sinigrin), myrosinase, glucoerucin, and limonene, which act as antimicrobial agents by damaging bacterial cell membranes, inhibition of enzyme activity, interference with DNA/RNA synthesis, and inhibiting the growth of pathogenic microorganisms39. The findings Odo et al.51 revealed antibacterial activity of turmeric extracts against Bacillus species, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. The antimicrobial potentials of date fruit gel, mustard seeds, and turmeric extracts are primarily ascribed to their natural bioactive compounds, which contribute to their widespread uses in the food industry as culinary ingredients.

Phenolics, flavonoid contents and antioxidant potentials of fortified plant-based milk

Figure 3 shows the total phenolic and flavonoid contents of plant-based milk containing extracts from date fruit, mustard seed and turmeric. The highest phenolic and flavonoid contents of 4.60 mg (GAE)/mL and 1.90 (mg QE)/mL, respectively, was obtained in 97%TM + 3%MS. The phenolic and flavonoid contents increased in plant-based milk supplemented with extract from mustard seeds, turmeric, and date fruit when compared to plant milk without the plant extracts. Plant-based milk contains a variety of bioactive compounds such as alpha-tocopherol, isoflavones, lauric acid, phytosterols—particularly β-sitosterol and γ-oryzanol, sesamin, sesamolin, sesamino, and others5. The incorporation of turmeric, date fruit, and mustard seeds further enhances the bioactive profile of plant-based milk, presenting a promising strategy for the development of functional foods. Milk from coconut, soy bean and tiger nut scavenged DPPH with values of 50.40%, 61.30%, and 67.40%, respectively. Plant milk enriched with the extracts from date fruit, mustard seed, and turmeric exhibited better (P ≤ 0.05) scavenging activity, ranging from 73.80 to 89.10% against DPPH with the highest activity observed in the 97%TM + 3%MS. Plant milk (97%SM + 3%MS) exhibited better ferric reducing antioxidant power of 305.10 µmol TE/mL, followed by 97%SM + 3%TR and 97%TM + 3%MS with values of 274.40 µmol TE/mL and 273.50 µmol TE/mL, respectively. Soy milk containing extract from mustard seeds (97%SM + 3%MS) scavenged ABTS with the highest value (P ≤ 0.05) of 131.30 µmol TE/mL. The enhanced antioxidant activity observed in the supplemented plant-based milk can be attributed to the bioactive compounds present in the extracts of date fruit, mustard seed, and turmeric. These bioactive compounds exhibit free radical scavenging, metal chelation, and lipid peroxidation inhibition activities. Findings of Tong et al.52 revealed valuable insights into the antioxidant potentials of plant-based milk as alternative sources to suppress oxidative stress to maximizing health benefits.

Fig. 3.

Fig. 3

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(a) Total phenolic and flavonoid contents, (b) DPPH scavenging (%), (c) Ferric reducing antioxidant power (FRAP), and (d) 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity of plant- based milk with and without extracts from date fruit, mustard seeds and turmeric. Error bar is standard deviation. CM: Coconut milk, SM: soy milk, TM: tiger nut milk, DG: date fruit gel, MS: mustard seed extract, TR: turmeric extract.

Conclusion

The supplementation of plant-based milk with extract from date fruit , mustard seed, or turmeric enhances nutritional value, supports probiotics growth, increase antioxidant activity, and suppressed the growth of diarrheagenic E. coli. Inclusion of medicinal plants into plant-based milk is an excellent source of protein, crude fiber, and mineral contents. The plant extracts in plant-milk are richer in phytochemicals with immense health benefits to man. The antioxidant and antimicrobial activities displayed by the fortified plant-based milk are likely attributed to the presence of polyphenols, flavonoids, and other bioactive compounds. The addition of these bioactive compounds into plant-based milk enhances recorded free radical scavenging, metal chelation, membrane disruption, and inhibition of microbial enzymes and oxidative stress pathways. The outcome of the study is offering guidance for the food industry to enhance the nutritional profile of these beverages by fortifying it with extracted bioactive compounds from medicinal spices and herbal plants. While our study focused on nutrient contents, and bio-functional activities, future research should explore the underlying molecular mechanisms responsible for these effects and evaluate the long-term health impacts of consuming fortified plant-based milk.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary Material 1. (249KB, docx)

Author contributions

C.O.O., O.A.O., K.O.K., O.O.O.: conceived and designed the experiment. C.O.O., O.A.O., K.O.K.: performed the experiments. C.O.O., K.O.K., O.O.O., F.O.A.: contributed reagents, materials, analysis tools or data. C.O.O., O.A.O., K.O.K.: drafted the manuscript. C.O.O., O.O.O., F.O.A.: reviewed and edited the manuscript. All the authorshave read and approved the final manuscript.

Funding

No fund received.

Data availability

The data supporting the findings of this study are available within the article and in the supplementary file.

Declarations

Competing interests

The authors declare no competing interests.

Footnotes

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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Supplementary Materials

Supplementary Material 1. (249KB, docx)

Data Availability Statement

The data supporting the findings of this study are available within the article and in the supplementary file.

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