Nutritional Profiling of Wild Edible Plants: Quantification of Macronutrients and Minerals via Energy Dispersive X-ray (EDX) Fluorescence Spectroscopy

Perfil nutricional de plantas comestíveis silvestres: quantificação de macronutrientes e minerais por espectroscopia de fluorescência de raios X dispersiva de energia (EDX)

Abstract

The wild edible plants offer a valuable resource for food and human nutrition. The current study aimed to analyze the proximate composition and ethno-medicinal profiles of twenty selected wild edible plants from the Malakand District, Pakistan. The proximate composition (moisture, ash, crude protein, crude fat, and dietary fiber) was analyzed following Association of Official Agricultural Chemists (AOAC) protocols. Ethno-medicinal profiles were obtained through semi-structured interviews with local informants. Mineral elements were analyzed using Energy Dispersive X-ray Fluorescence (EDX) Spectroscopy. The normalized variation matrix and centered log ratio (CLR) biplot were used to assess linear associations between nutritional components. The results revealed significant variations in the nutritional components and mineral contents among the studied plant species. Higher crude carbohydrates (64.48%) were scrutinized in Ziziphus jujuba, followed by proteins (19.90%) in leaves of Solanum nigrum, fiber (19.50%) in Caralluma tuberculata and fats (6.12%) in Mentha spicata. A maximum energy value of 332.34 Kcal/100g was calculated for leaves of Solanum nigrum. The concentrations of carbon (C) in Mentha longifolia were (60.20%), followed by oxygen (46.06%) in Ziziphus jujuba, potassium (10.80%) in Rumex dentatusand nitrogen (7.98%) in Nasturtium officinale. The centered log ratio biplot confirms strong relationships between moisture, energy, carbohydrates, fibers, and ash content, while protein and fat concentrations exhibit a separate pattern. The findings provide valuable insights into the compositional data analysis of edible plant species, suggesting interdependencies among carbohydrate, energy, moisture content, and fibers. Additionally, the results of our current study showed that all of these selected wild edible plants have the potential to provide humans with important macronutrients and elements.

Keywords:
Wild edible plants; Proximate analysis; Mineral composition; Compositional data analysis (CoDa); Energy Dispersive X-ray Fluorescence (EDX) Spectroscopy; Human Health

Resumo

As plantas comestíveis selvagens oferecem um recurso valioso para a alimentação e nutrição humana. O presente estudo teve como objetivo analisar a composição centesimal e os perfis etnomedicinais de vinte plantas comestíveis silvestres selecionadas do distrito de Malakand, Paquistão. A composição centesimal (umidade, cinzas, proteína bruta, gordura bruta e fibra alimentar) foi analisada seguindo protocolos da Association of Official Agricultural Chemists (AOAC). Os perfis etnomedicinais foram obtidos por meio de entrevistas semiestruturadas com informantes locais. Os elementos minerais foram analisados por espectroscopia de fluorescência de raios X por dispersão de energia (EDX). A matriz de variação normalizada e o gráfico biplot de razão logarítmica centrada (CLR) foram utilizados para avaliar associações lineares entre componentes nutricionais. Os resultados revelaram variações significativas nos componentes nutricionais e nos teores minerais entre as espécies vegetais estudadas. Carboidratos brutos mais elevados (64,48%) foram identificados em Ziziphus jujuba, seguidos por proteínas (19,90%) em folhas de Solanum nigrum, fibras (19,50%) em Caralluma tuberculata e gorduras (6,12%) em Mentha spicata. Um valor energético máximo de 332,34 Kcal/100g foi calculado para as folhas de Solanum nigrum. As concentrações de carbono (C) em Mentha longifolia foram (60,20%), seguidas de oxigênio (46,06%) em Ziziphus jujuba, potássio (10,80%) em Rumex dentatus e nitrogênio (7,98%) em Nasturtium officinale. O gráfico biplot de razão logarítmica centrada confirma fortes relações entre umidade, energia, carboidratos, fibras e teor de cinzas, enquanto as concentrações de proteína e gordura exibem um padrão separado. As descobertas fornecem informações valiosas sobre a análise de dados de composição de espécies vegetais comestíveis, sugerindo interdependências entre carboidrato, energia, teor de umidade e fibras. Além disso, os resultados do nosso estudo atual mostraram que todas essas plantas comestíveis silvestres selecionadas têm o potencial de fornecer aos seres humanos macronutrientes e elementos importantes.

Palavras-chave:
plantas silvestres comestíveis; análise centesimal; composição mineral; análise de dados composicionais (CoDa); espectroscopia de fluorescência de raios X por dispersão de energia (EDX); Saúde Humana

1. Introduction

The rapidly growing world population has increased the need for food and nutrition and lead to the depletion of available natural land resources (Wang, 2022). In recent years, there has been growing interest in the investigation of wild edible plants due to their potential as alternative sources of food and medicine.

Wild edible plants have been integral to the diets and medicinal practices of various cultures for centuries. These plants not only offer a rich source of nutrients but also hold significant ethnobotanical value, serving various traditional health and providing valuable nutrients and bioactive compounds (Anusiya et al., 2021; Motti, 2022). The world nutritionists strive to know and identify inexpensive and readily available food substitutes to support national and international governments and agencies such as the Food and Agriculture Organization (Patwardhan and Paranthaman, 2021). District Malakand, situated in the Khyber Pakhtunkhwa province of Pakistan, is known for its rich biodiversity, with a wide range of wild edible plants flourishing in its diverse ecosystems (Khan et al., 2019).

Despite the recognized importance of wild edible plants, there are significant gaps in how their nutritional composition aligns with their traditional uses, particularly in regions with rich biodiversity like Malakand (Ibrahim et al., 2023). Most existing studies have focused on the identification and traditional uses of these plants but lack comprehensive quantitative analysis of their macronutrient and mineral content (Zabihullah et al., 2006; Barkatullah and Ibrar., 2011; Barkatullah et al., 2015; Khan et al., 2019). Quantitative analysis of macromolecules, including proteins, carbohydrates, and lipids, provides valuable insights into the nutritional quality of wild edible plants (Zhong et al., 2022). Proteins are essential macromolecules that play a crucial role in building and repairing tissues, while carbohydrates serve as the primary energy source for human metabolism (Thakur et al., 2023). Lipids, on the other hand, are important for energy storage and the absorption of fat-soluble vitamins (Campbell, 2017). The quality of these crucial molecules in plants are the key aspects for the grouping and taxonomy as well as improvements of plants for future planning (Barboza et al., 2009). Understanding the macromolecular composition of wild edible plants can help identify their nutritional value and evaluate their potential contribution to combating malnutrition and food insecurity (Shaheen et al., 2017; Thakur et al., 2022).

Furthermore, elemental investigation of wild edible plants offers insights into their mineral composition (Drava et al., 2020). Minerals are vital for maintaining various physiological functions in the human body, such as bone formation, nerve transmission, and enzyme activity (Bharti et al., 2021). Analyzing the elemental content of wild edible plants can provide valuable information about their mineral profile and assess their potential as natural sources of essential minerals (Radha et al., 2021). In many developing and poor countries of the world, rural people use and rely only on wild edible fruits available to them as they cannot afford to buy commercial fruits (Jamnadass et al., 2011; Dalu et al., 2021). Research studies have shown that some edible wild fruits have higher nutritional value than the commercial fruits available in the market (Sundriyal and Sundriyal, 2004; Egea et al., 2010; Seal, 2011; Biswas et al., 2022).

The unique ecological characteristics of Malakand District make it an ideal site for the study of wild edible plants as the diverse topography of the region, ranging from mountains to plains, contributes to a rich and varied flora (Barkatullah and Ibrar., 2011; Khan et al., 2017). Additionally, the local communities in this area have a deep-rooted connection with wild edible plants, utilizing them for traditional medicinal purposes and as food resources (Murad et al., 2011, 2012). Therefore, this research aims to fill the gaps in the current understanding by conducting a comprehensive quantitative analysis of macromolecules and mineral elements of selected wild edible plants in Malakand District, using energy dispersive X-ray spectroscopy. This study is crucial for evaluating their nutritional potential and supporting efforts to combat food insecurity and malnutrition.

2. Materials and Methods

2.1. Selection criteria for nutritional profiling of wild edible plants in Malakand

Plants were selected to represent a diverse range of ecological niches within Malakand, encompassing various altitudes, soil types, and climatic conditions (Figure 1) to provide a comprehensive nutritional assessment (Ibrahim et al., 2023). Traditional knowledge and local usage guided the selection focusing on plants commonly used by local communities for food and medicine (Table 1 ). The rich biodiversity of Malakand ensured a variety of species, capturing the nutritional diversity of the region (Jan et al., 2012). Plants were chosen for their known or potential nutritional value, particularly those rich in essential macronutrients and minerals (Zabihullah et al., 2006; Murad et al., 2011; Khan et al., 2017, 2019). Availability and accessibility were also considered to facilitate sample collection, while the selection aimed to address research gaps by including plants that had not been extensively studied, ensuring that the findings are relevant and comprehensive.

Figure 1
Shows map of the study area with sampling sites in District Malakand.

Table 1
Wild Edible Plants and their Ethnobotanical uses.

S.No	Plant name	Loal name	Family Name	Ethnobotanical uses
1	Ammaranthus viridis L.	Chalwai	Ammaranthaceae	Fresh Leaves are boiled as vegetables and used as a laxative.
2	Asparagus officinalis L.	Tendonai	Asparagaceae	Young shoots are fried and used as vegetables.
3	Bauhinia variegata (L.) Benth.	Kulyar	Fabaceae	Young floral leaves are cooked as vegetables. The flowers and buds are also used in pickles.
4	Caralluma tuberculata N.E.Brown	Pamankai	Asclepiadaceae	Succulent stem is cooked as vegetables. The juice extracted from stem and roots is used to cure jaundice and diabetes.
5	Chenopodium album L.	Saarmey	Chenopodiaceae	The leaves are cooked as vegetables. It is also used in the form of pot herb in piles.
6	Foeniculum vulgare Mill.	Kaga	Apiaceae	Young shoots and leaves are used as salad. The dried fruits are used as carminative and digestive.
7	Malva neglectaWallr.	Paneerak	Malvaceae	Young leaves are cooked as vegetables. Also used to cure cough, respiratory and digestive problems.
8	Medicago polymorpha L.	Peeshtaray	Fabaceae	The leaves and young shoots are cooked as vegetables. Also used as fodder for cattle’s. It is also used medicinally to cure diabetes and skin infections.
9	Meliolotus indicus (L.) All.	Leewanai	Fabaceae	The leaves are cooked as vegetables. The seeds are made in to a gruel and used to cure bowl complaints and as emollient. It is also used as fodder.
10	Mentha longifolia (L.) Huds.	Enalai	Lamiaceae	Young leaves are chewed raw as salad. The herbal tea made from the leaves are used to treat diarrhea and vomiting.
11	Mentha spicata L.	Podina	Lamiaceae	Young leaves are used as salad. The herbal tea made from the leaves are used to treat abdominal pain and indigestion.
12	Morus alba L.	Toot	Moraceae	The ripe fruits are eaten fresh or in dried form. The dried fruits are used as tonic. The leaves are used as fodder.
13	Nasturtium officinale W.T.Aiton	Tarmeera	Brassicaceae	The young leaves and shoots are used as vegetables. The plant is medicinally used as diuretic and expectorant.
14	Oxalis corniculata L.	Treewakai	Oxalidaceae	The leaves and young shoots are used as vegetables. Medicinally the plant is used to cure intestinal worms and promote urination.
15	Portulaca oleracea L.	Warkharai	Portulaceae	The leaves and stem are cooked as vegetables. Locally the plant is also used to cure constipation and as emollient.
16	Rumex dentatus L.	Shalkhai	Polygonaceae	The leaves are used as vegetables. Plant is also used as a fodder. The ethno- pharmacological uses of the plant are as diuretic and refrigerant.
17	Rumex hastatus Don.	Tarookai	Polygonaceae	The leaves are used as vegetables. Locally the plant is also used as astringent and for skin irritations.
18	Solanum nigrum L. (Leaves)	Kachmachu	Solanaceae	The leaves are used as vegetables. The leaves are also used as anti-inflammatory and as diuretic.
19	Solanum nigrum L. (Fruits)	Kachmachu	Solanaceae	The ripened fruits are eaten as raw. The fruits are used to dispel phlegm and cure diarrhea.
20	Ziziphus jujuba Mill.	Beera	Rhamnaceae	The ripened fruits are eaten as raw. The leaves are used as fodder. The ripened fruits are medicinally used as emollient, digestive and sedative.

2.2. Sample collection and preparation

Field inventories for the collection of edible wild plants were carried out from March 2018 to September 2020 in the district of Malakand, Pakistan (Figure 1). A total of twenty different plant species were collected and analyzed based on their traditional use as edible plants and their availability in the region. Proper care was taken to collect representative samples of each plant species, including leaves, stems, and fruits. The collected plant specimen was identified with the help of existing literature and flora of Pakistan (Abbas et al., 2017). The collected plant samples were thoroughly washed with distilled water to remove any contaminants or dirt. Fresh plant samples or their parts were used to determine the moisture content. The samples were then air-dried at room temperature to a constant weight (Hussain et al., 2018). Dried samples were ground to a fine powder using an electric grinder and stored in air tight bottles at room temperature for further analysis (Khan et al., 2021a).

For the ethno-medicinal profile, semi-structured interviews were conducted with local informants to document traditional uses of each plant species. Data were analyzed to correlate proximate composition with ethno-medicinal applications (Dewi et al., 2023).

2.3. Proximate nutritional analysis

The proximate nutritional analysis of the selected plant samples was performed in accordance with the official methods of the Association of Official Agricultural Chemists (AOAC International, 2019; Sharma et al., 2020).

2.4. Determination of moisture content

The moisture content of fresh plant samples was determined by the oven-dried method at 105 °C using the official method of (AOAC International, 2019).

The moisture content was calculated using the following Equation 1,

$Moisturecontent\left(\%\right)=\left(W2-W3/W2-W1\right)\times 100$ (1)

Where W1is weight of empty crucible, W2 is weight of sample and crucible, and W3 is weight of sample with contents after drying.

2.5. Determination of ash content

The ash content (%) was determined by burning the dried plant samples in a muffle furnace at 550 °C for 4 hours (AOAC International,2019).

The formula used to determine the ash content (%) is given below (Equation 2),

$Ash\left(\%\right)=\left(W2-W1/W3\right)x100$ (2)

where W1 is the weight of the crucible, W2 is the weight of the sample and crucible after drying, and W3 is the weight of the sample on dry weight basis.

2.6 Determination of crude fats

The crude lipid content in the tested samples was determined with a Soxhlet apparatus using the petroleum ether (40 – 60 ⁰C) solvent extraction method (Ali et al., 2023). The calculation was done using the given formula (Equation 3),

$Crudefat\left(\%\right)=\left(Wf/Ws\right)x100$ (3)

where Wf is weight of fats in sample, and Ws is weight of sample.

2.7. Determination of crude proteins

The determination of crude proteins was carried out using a macro-Kjeldahl method (Labconco, 2008; Wang et al., 2016).

The nitrogen fraction was determined using the formula below (Equation 4),

$\left(N\%\right)=\left(S-B\right)\times N\times 0.014\times D/Weightofsamplex100$ (4)

Where D is Dilution factor, T is titration value (S – B), W is Weight of sample, and 0.014 is constant value, Crude protein % = N × (6.25 factor).

2.8. Determination of crude fibers

The crude fiber content in the samples was determined using the methods of (AOAC International, 2019). The following formula was used to calculate the crude fiber content (Equation 5),

$Crudefibers\left(\%\right)=\left(W1-W2/W3\right)\times 100$ (5)

where W1 is weight of crucible before drying, W2 is weight of crucible after drying and W3 is weight of plant sample.

2.9. Determination of carbohydrates

To calculate carbohydrates, the sum total of ash (%), crude fibers (%) crude proteins (%) and crude fats (%) were from subtracting values from 100 (Ali et al., 2023) (Equation 6).

$Carbohydrates\%=100–\left(\begin{array}{c}Ash\%+crudefibers\%+\\ crudeproteins\%+crudefats\%\end{array}\right)$ (6)

2.10. Determination of Energy value (Ev)

The energy values (kcal/100 g) of each sample were calculated by multiplying the values of carbohydrate, lipid and protein by a factor of 4, 9, and 4 respectively (Okwu and Morah 2004; Hussain et al., 2010).

2.11. Energy dispersive X-ray fluorescence analysis

Two gram dried plant material of the selected plant parts were initially pulverized for 10 minutes in a planetary ball mill (Pulverisette 7; Fritsch GmbH, Idar-Oberstein, Germany) at 450 rpm (135 m/s² or 13.8 G), followed by pressing into 32-mm diameter pellets using a hydraulic press at 20 tons of pressure (Specac, Kent, UK) (Matsunami et al., 2010). Elemental analysis of the prepared samples was carried out using an energy dispersive X-ray fluorescence spectrometer (EDX-7000, Na-U, Shimadzu, Japan) at the Centralized Resource Laboratory (CRL), University of Peshawar, Pakistan (Khan et al., 2021a; Nyakuma et al., 2021). The instrument is equipped with a Rhodium (Rh) target X-ray tube and a high-performance silicon drift detector (SDD), operating at a maximum of 50 kV and 1000 μA, and controlled by PCEDX-Navi software. The elemental composition of all samples was detected under an air-based atmosphere. Analytes were assessed using a 10 mm diameter collimator with a live acquisition time of 60 seconds (Bilo et al., 2015; Nyakuma et al., 2021). To ensure the accuracy and reliability of the EDXRF results, the instrument was calibrated using the standards 1570a (Spinach leaves), CTA OTL1 (Tobacco leaves) and 1573a (Tomato leaves) (Kulal et al., 2020). The samples were analyzed in triplicates for consistency. Strict protocols for sample preparation were followed to maintain consistency, and the analysis was conducted in a controlled environment to minimize external factors (Khan et al., 2021a).

2.12. Statistical analysis

Descriptive statistical analysis, focusing on the proximate nutritional and elemental analysis, was conducted utilizing the SPSS statistical package (version 25) and Excel 2016 software. To investigate the linear relationship between the proximate nutritional variables, Compositional Data Analysis (CoDa) techniques, including the centered log-ratio transformation (CLR), were applied to the raw data (Khan et al., 2022). Log ratio-variation matrix, CLR covariance biplots and a CoDa-dendrogram were constructed to identify the connections between variables (Egozcue and Pawlowsky-Glahn, 2019; Boente et al., 2022). The CoDaPack software and the R package 'compositions' were utilized for these computations (van den Boogaart and Tolosana-Delgado, 2013; Thio-Henestrosa and Comas 2016).

3. Results

3.1. Integrated analysis of the proximate composition and ethno-medicinal profile of plants

The proximate compositions of wild edible plant species from various locations in Malakand District were presented as mean ± standard deviation (%) in Table 2. The plant species exhibited significant variation in average moisture content. The moisture content ranged from 78.29 ± 0.4% to 90.74 ± 0.5%. The highest average moisture contents were recorded for Portulaca oleracea L. (90.74 ± 0.5%), Rumex dentatus L. (90.06 ± 0.4%), and Oxalis corniculata L. (90.02 ± 0.02%). In contrast, the lowest average moisture content was found in Chenopodium album L. (78.29 ± 0.4%) (Table 2). The leaves and stems of P. oleracea are utilized as vegetables and emollients (Table 1). The high moisture content aids in maintaining the plant’s freshness and nutritional value. Similarly, the leaves of R. dentatus are used as vegetables and fodder. This high moisture content enhances its efficacy as a diuretic and refrigerant (Table 1). The leaves and young shoots of O. corniculata are consumed as vegetables and used medicinally to treat intestinal worms and promote urination. Therefore, the high moisture content supports their medicinal applications.

Table 2
Proximate Nutritional Analysis of Wild Edible Plants in District Malakand.

Plant Name	Part used	MC (%)	DM (%)	Ash%	Fib (%)	CF (%)	CP (%)	Carb (%)	GEV (kcal/100 g)
Ammaranthus viridis L.	Leaves	79.31±0.3	20.69±0.1	17.59±0.02	15.19±0.3	3.01±0.02	6.85±0.02	48.36±0.3	247.93±0.4
Asparagus officinalis L.	Shoots	89.11±0.2	10.89±0.5	8.69±0.2	16.20±0.3	4.01±0.01	7.03±0.03	64.07±0.2	320.49±0.5
Bauhinia variegata (L.) Benth.	Floral leaves	79.89±0.3	20.11±0.02	10.88±0.04	12.59±0.4	3.99±0.03	14.86±0.02	57.68±0.4	326.07±0.6
Caralluma tuberculata N.E.Br	Succulent stem	83.52±0.2	16.48±0.2	15.30±0.3	19.50±0.4	2.01 ±0.03	8.06±0.02	55.13±0.2	270.85±0.2
Chenopodium album L.	Leaves	78.29±0.4	21.71±0.2	20.89±0.2	15.51±0.1	2.98±0.02	10.68±0.02	49.94±0.3	269.30±0.3
Foeniculum vulgare Mill.	Leaves	84.75±0.02	15.25±0.1	11.65±0.3	19.30±0.03	2.05±0.04	7.22±0.4	59.78±0.6	286.45±0.4
Malva neglecta Wallr.	Leaves	79.59±0.3	20.41±0.2	20.08±0.02	16.49±0.04	3.12±0.2	10.88±0.2	50.57±0.5	273.88±0.2
Medicago polymorpha L.	Leaves	79.34±0.4	20.66±0.03	10.53±0.2	15.58±0.3	3.02±0.3	11.49±0.03	59.38±0.6	310.66±0.4
Meliolotus indicus (L.) All.	Leaves	84.45±0.2	15.55±0.3	14.32±0.02	18.01±0.03	2.97±0.1	16.31±0.03	52.61±0.3	302.41±0.3
Mentha longifolia (L.) Huds.	Leaves	80.54±0.2	19.46±0.1	19.31±0.2	10.41±0.05	6.05±0.02	8.02±0.01	56.21±0.4	311.37±0.2
Mentha spicata L.	Leaves	79.83±0.02	20.17±0.2	19.94±0.3	7.88±0.03	6.12±0.03	8.02± 0.3	58.04±0.4	319.32±0.5
Morus alba L.	Fruit	84.58±0.4	15.44±0.02	16.05±0.3	13.51±0.2	2.02±0.02	6.29±0.04	62.14±0.3	291.81±0.1
Nasturtium officinale W.T.Aiton	Leaves	89.31±0.3	10.69±0.02	15.66±0.2	13.51±0.4	4.05±0.04	7.02±0.02	59.76±0.03	303.57±0.2
Oxalis corniculata L.	Leaves	90.02±0.02	09.98±0.02	12.65±0.4	15.30±0.2	2.99±0.01	8.11±0.3	60.95±0.04	303.15±0.3
Portulaca oleracea L.	Leaves/shoot	90.74±0.5	9.26±0.2	15.64±0.2	16.22±0.3	5.01±0.3	21.05±0.4	42.08±0.3	297.43±0.4
Rumex dentatus L.	Leaves	90.06±0.4	9.94±0.04	17.05±0.3	13.06±0.4	3.01±0.1	12.07±0.03	54.81±0.6	294.61±0.3
Rumex hastatus Don.	Leaves	84.32±0.4	15.68±0.02	15.71±0.03	14.05±0.3	5.10±0.03	9.65±0.3	55.49±0.03	306.46±0.1
Solanum nigrum L.	Fruits	80.35±0.03	19.65±0.3	11.95±0.02	10.20±0.02	4.05±0.4	19.64±0.3	54.16±0.4	331.65±0.3
Solanum nigrum L.	Leaves	79.70±0.03	20.30±0.3	13.07±0.2	10.12±0.2	5.02±0.2	19.90±0.3	51.89±0.3	332.34±0.6
Ziziphus jujuba Mill.	Fruit	80.30±0.2	19.70±0.3	9.16±0.2	14.65±0.03	2.98±0.2	8.73±0.2	64.48±0.02	319.66±0.3

• Key: MC (Moisture content), DM (Dry matter), CP (Crude proteins), CF (Crude Fats), Carb (Carbohydrates), Fib (Fibers), GEV (Gross Energy value).

C. album cooked leaves are used in remedies for piles and the lower moisture content contributes to its higher dry matter, enhancing its therapeutic properties.

The dry matter content values varied from 9.26±0.2% to 21.71± 0.2%. The highest dry matter was found in the leaves of Chenopodium album (21.71± 0.2%) followed by Ammaranthus viridis (20.69 ± 0.1%) and Medicago polymorpha (20.66 ± 0.03%). The lowest dry matter content of 9.26±0.2% observed in the leaves of Portulaca oleracea (Table 2). The high dry matter content of C. album enhances its effectiveness in therapeutic applications due to the concentration of active compounds. Likewise, A. viridis L. is traditionally consumed as a laxative. Our nutritional analysis reveals that it is high in dietary fiber, which supports its use in promoting digestive health (Table 1). M. polymorpha leaves and young shoots are used as vegetables and fodder, and has additional medicinal uses for diabetes and skin infections (Table 1). The elevated dry matter supports its role in nutrition and medicine. The lower dry matter content of P. oleracea indicates higher water content, which contributes to its use in alleviating constipation and providing freshness in culinary applications.

The ash content measures the mineral content in the food material (Hussain et al., 2010). The ash contents for the collected wild edible plants ranged between 20.89 ± 0.2% and 8.69±0.2% (Table 2). The ash content was high in Chenopodium album (20.89 ± 0.2%) followed by Malva neglecta (20.08± 0.02%), and Mentha spicata (19.94± 0.3%). A minimum ash content of 8.69±0.2% was reported for Asparagus officinalis (Table 2). The high ash content in C. album signifies a rich mineral profile, which supports its medicinal efficacy. Similarly, the young leaves of M. neglecta are utilized as vegetables and for treating coughs, respiratory issues, and digestive complaints (Table 1). The elevated ash content reflects its substantial mineral content beneficial for these therapeutic uses. The young leaves of M. spicata are used for salads and herbal tea to alleviate abdominal pain and indigestion (Table 1). The high ash content indicates its rich mineral content, enhancing its digestive benefits. A. officinalis young shoots are fried and consumed as vegetables (Table 1). The lower ash content suggests a lower mineral concentration, which might influence its nutritional profile in culinary applications.

Accordint to (Tura et al., 2023), dietary fiber stimulates growth and safeguards the advantageous gut microbiota. Caralluma tuberculata exhibited the highest level of crude fiber content of 19.50 ± 0.4%, followed by Foeniculum vulgare (19.30 ± 0.03%) and Meliolotus indicus (18.01 ± 0.03%), (Table 2) respectively. C. tuberculata succulent stem is used as a vegetable, and its juice treats jaundice and diabetes (Table 1). The high fiber content supports its use in digestive health and weight management. Similarly, F. vulgare young shoots and leaves are used in salads, while its dried fruits serve as carminative and digestive aids (Table 1). This shows high levels of carminative essential oils and minerals like magnesium, which are known to aid digestion. In addition, M. indicus cooked leaves are used as vegetables, and its seeds are made into gruel to address bowel complaints (Table 1). The high fiber content complements its use as an emollient and digestive aid.

Mentha spicata was observed with the lowest fat content of 7.88±0.03%. The lipid content is of crucial importance in nutritional analysis as an energy source (Ganogpichayagrai and Suksaard, 2020). In the present study, significant variations in fat compositions were observed among the plants (Table 2). A minimum fat content of 2.01 ± 0.03% was observed in succulent stem of Caralluma tuberculata. Mean maximum crude fat content of 6.12± 0.03% were investigated in leaves of Mentha spicata followed by Mentha longifolia (6.05±0.02%), and Rumaxhastatus (5.10 ±0.03%), respectively. All the twenty plant species included in our study exhibited crude fat content of above 2%. M. spicata young leaves are used for salads and herbal tea to relieve abdominal pain and indigestion (Table 1). Similarly, M. longifolia young leaves are eaten raw as salad, and its herbal tea is used for treating diarrhea and vomiting (Table 1). The higher fat content supports its role in providing energy and nutritional benefits. The R. hastatus leaves are utilized as vegetables and applied locally as an astringent and for skin irritations (Table 1). Their moderate fat content supports its use in local culinary and medicinal practices. Whereas, the low fat content of C. tuberculata aligns with its use in traditional medicine and dietary practices.

Protein is crucial for growth, body development, tissue repair, enzyme and hormone production, and overall optimal functioning (Ornitz and Itoh, 2015). In the current study, Portulaca oleracea was found to possess the highest protein content of 21.05 ± 0.4% followed by Solanum nigrum (19.90 ± 0.3%). Morus alba exhibited the lowest total protein content of 6.29±0.04% (Table 2). Generally, the majority of the plants examined in this study have protein levels higher than 6.29 g/100 g. The high protein content of P. oleracea supports its role as a significant dietary source in traditional cuisine (Table 1). S. nigrum leaves are used as vegetables with anti-inflammatory and diuretic properties, and its ripe fruits help dispel phlegm and treat diarrhea (Table 1). The substantial protein content enhances its nutritional value and therapeutic benefits. Similarly, M. alba provides ripe fruits consumed fresh or dried, with dried fruits serving as a tonic and leaves as fodder (Table 1). Despite the lower protein content, it remains valuable for its medicinal and nutritional properties.

The main source of energy in the human body are carbohydrates. The analysis of the carbohydrate content in wild edible plants revealed notable variations. The average higher carbohydrate content of 64.48±0.2% were observed in Ziziphus jujuba followed by Asparagus officinalis (64.07±0.02%) and Morus alba (62.14±0.3%), respectively. The average minimum carbohydrate content of 42.08±0.3% was reported in Portulaca oleracea. The Z. jujuba ripe fruits are consumed raw, with its leaves used as fodder (Table 1). The high carbohydrate content contributes to its role as a significant energy source in traditional diets, supporting its medicinal uses as an emollient, digestive, and sedative. (Table 1). Likewise, the high carbohydrate content in A. officinalis and M. alba content supports its role as an energy source and tonic. Despite the lower carbohydrate content of P. oleracea, it remains significant for its traditional uses.

In the current study, the gross energy value of the wild edible plants ranged between 247.93±0.4 and 332.34 ± 0.6 Kcal/100g (Table 2). The highest energy values of 332.34 ± 0.6 Kcal/100g and 326.07 ± 0.6 were calculated for leaves of Solanum nigrum and fruits of Bauhinia variegata. The lowest energy value of 247.93±0.4 Kcal/100g was obtained for Ammaranthus viridis leaves. The high energy content of S. nigrum supports its use in traditional diets as a vegetable with anti-inflammatory and diuretic properties, and its ripe fruits aid in dispelling phlegm and treating diarrhea (Table 1). The substantial energy content of B. variegata aligns with its use in cooking and pickling, providing both nutritional value and medicinal benefits (Table 1). Despite the lower energy content of A. viridis, the plant remains significant for its nutritional and laxative properties (Table 1).

3.2. Looking for association between the variables of proximate composition

In the current study, the smallest contribution from carbohydrates to energy content (0.01), moisture content (0.02), dry matter (0.06), fibers (0.06), and energy to moisture content (0.01). Similarly, smaller contributions, less than 0.2, from fibers to moisture content (0.03), ash to moisture content (0.05) and ash to energy content (0.07) (Table 3). The relatively large normalized variation in the logarithmic ratios of fats with dry matter (0.27), fibers (0.23), and proteins with dry matter (0.24) separated fats, proteins, and dry matter first in the cluster of the dendrogram. Similarly, proportional logarithmic ratios of moisture content to carbohydrates (0.02), energy (0.01), and ash content to fibers (0.11), keep them in their respective clusters (Figure 1). The result of Table 3 is verified by centered log ratio (CLR) biplot, as depicted in Figure 2, which explains 66.4% of the total variability in the data. The close proximity of the vertices in the biplot implies that these variables are associated and tend to change together (Figure 2). Figure 3 further confirmed the results of table 3 and figure 2 as the closely related proximate composition variables fall within same clusters.

Table 3
The normalized variation matrix of Table 2 shows the linear relationship between variables.

Variables (clr)	Fats	Proteins	Carbohydrates	Ash	Fibers	Energy	Moisture	Dry matter
Fats
Proteins	0.14
Carbohydrates	0.21	0.21
Ash	0.15	0.18	0.10
Fibers	0.23	0.20	0.06	0.11
Energy	0.13	0.13	0.01	0.07	0.06
Moisture	0.14	0.13	0.02	0.05	0.03	0.01
Dry matter	0.27	0.24	0.06	0.14	0.15	0.07	0.10

Figure 2
Clr-biplot of the proximate analysis of wild edible plants shows the relationship between variables and the observations.

Figure 3
The hierarchical cluster dendrogram of the association of the proximate composition of wild edible plants reproduced by the R packages ‘compositions’ and ‘hclust’ with the Ward method.

3.3. Macronutrients and minerals composition of the wild edible plants.

Table 4 presents the analysis of various macronutrients and minerals found in samples taken from different altitude ranges. The observed macro elements were carbon (C), calcium (Ca), magnesium (Mg), potassium (K), sodium (Na), nitrogen (N), phosphorus (P), and sulfur (S), along with microelements like iron (Fe), copper (Cu), and silicon (Si). Carbon (C) was identified as the predominant macro element, with concentrations ranging from 47.09% to 60.20% (Table 4). The highest carbon concentration was observed in Mentha longifolia (60.20%), followed by Medicago polymorpha (58.27%) and Bauhinia variegata (57.11%). Similarly, Ziziphus jujuba exhibited the highest oxygen concentration of 46.06%, Caralluma tuberculata (40.31%) and Morus alba (39.78%), respectively. The maximum nitrogen content was found in Nasturtium officinale (7.98%), followed by Asparagus officinalis (7.12%) and Melilotus indicus (7.04%). The highest potassium concentration was recorded in Rumex dentatus (10.80%), followed by Chenopodium album (9.07%) and Portulaca oleracea (7.05%) (Table 4).

Table 4
Mineral composition of the selected Wild Edible Plants in District Malakand.

Plant Names	Part used	C %	O %	N %	Mg %	Al %	Si %	P %	S %	K %	Ca %	Cl %	Na %	Fe %	Cu %
Ammaranthus viridis L.	Leaves	49.1	34.11	6.05	0.47	0.23	1.2	0.7	0.2	2.81	2.01	0.71	-	0.31	0.43
Asparagus officinalis L.	Shoots	54.14	35.3	7.12	0.21	0.08	0.31	0.51	0.41	1.31	0.2	0.41	-	-	-
Bauhinia variegata (L.) Benth.	Floral leaves	57.11	37.12	-	0.62	-	0.42	-	0.2	0.51	4.02	-	-	-	-
Caralluma tuberculata N.E.Br	Succulent stem	50.57	40.31	6.48	0.33	0.32	0.61	-	0.1	0.31	0.88	-	0.09	-	-
Chenopodium album L.	Leaves	51.44	30.3	5.1	0.62	0.41	0.79	0.32	0.21	9.07	0.3	0.42	-	0.31	0.71
Foeniculum vulgare Mill.	Leaves	53.05	37.81	-	0.31	0.39	0.61	0.3	0.81	1.99	2.97	0.63	0.42	-	0.71
Malva neglecta Wallr.	Leaves	54.16	34.6	-	0.8	0.24	0.75	0.31	0.79	2.98	2.99	0.71	0.51	0.21	0.95
Medicago polymorpha L.	Leaves	58.27	34.09	-	0.31	-	0.47	0.44	0.29	4.11	1.03	-	-	-	0.99
Meliolotus indicus (L.) All.	Leaves	51.21	33.51	7.04	0.42	0.62	2.01	0.32	0.31	0.81	3.01	-	-	0.74	-
Mentha longifolia (L.) Huds.	Leaves	60.2	29.28	-	0.21	0.41	0.82	-	0.43	3.02	4.01	0.81	-	0.41	0.4
Mentha spicata L.	Leaves	51.75	34.75	6.03	0.51	0.42	0.79	0.22	0.52	2.05	2.02	0.94	-	-	-
Morus alba L.	Fruit	50.24	39.78	6.31	0.21	0.23	0.4	0.22	0.2	2.2	0.21	-	-	-	-
Nasturtium officinale W.T.Aiton	Leaves	49.69	29.51	7.98	0.3	0.4	2.01	0.49	0.22	4.01	2.04	2.01	0.31	0.41	-
Oxalis corniculata L.	Leaves	50.22	32.76	-	0.81	2.01	5.02	-	0.23	3.04	2.01	0.32	-	2.03	0.62
Portulaca oleracea L.	Leaves/shoot	56.9	29.98	-	0.72	0.24	0.85	0.42	-	7.05	2.03	0.64	-	0.45	0.72
Rumex dentatus L.	Leaves	47.09	29.86	6.06	0.51	0.34	0.91	0.56	0.41	10.8	0.74	0.72	0.3	0.22	0.67
Rumex hastatus Don.	Leaves	53.17	37.65	-	0.82	0.21	0.53	0.31	0.3	3.21	2.05	0.84	-	-	0.91
Solanum nigrum L.	Fruits	53.26	34.3	7.01	0.31	0.21	0.2	0.32	-	3.1	0.63	0.34	0.32	-	-
Solanum nigrum L.	Leaves	54.76	29.61	5.12	0.54	0.51	2.11	0.34	0.42	2.12	3.06	0.87	-	0.54	-
Ziziphus jujuba Mill.	Fruit	47.26	46.06	5.13	-	-	-	0.22	-	0.89	0.21	0.23	-	-	-

• Key: C (Carbon), O (Oxygen), N (Nitrogen), Mg (Magnesium), Al (Aluminum), Si (Silicon), P (Phosphorus), S (Sulfur), K (Potassium), Ca (Calcium), Cl (Chlorine), Na (Sodium), Fe (Iron), Cu (Copper).

Calcium concentrations varied significantly among the edible wild plants, ranging from 0.20% to 4.02%. The highest calcium levels were found in Bauhinia variegata (4.02%) and Mentha longifolia (4.01%), while Asparagus officinalis showed the lowest value (0.20%). Chloride concentrations also varied, with values between 0.23% and 2.01%. Nasturtium officinale had the highest chloride content (2.01%), whereas Ziziphus jujuba had the lowest (0.23%). Concentrations of other mineral concentrations were as follows: magnesium ranged from 0.21% to 0.82%, sulphur from 0.10% to 0.81%, phosphorus from 0.22% to 0.70%, and sodium from 0.09% to 0.51% (Table 4).

The results of micro elements such as Si, Fe and Cu showed significant variations in accumulation among the studied plants. The silicon content ranged from 0.20% to 5.02% (Table 4). The lowest Si content was observed in Solanum nigrum fruits (0.20%), while Oxalis corniculata leaves exhibited the highest Si content (5.02%). The iron content varied between 0.21% and 2.03% (Table 4). Oxalis corniculata showed the highest Fe concentration (2.03%), whereas Malva neglecta had the lowest Fe content (0.21%). Copper concentrations were relatively low, ranging from 0.40% to 0.99% (Table 4). Mentha longifolia and Medicago polymorpha showed the lowest and highest Cu content, respectively. High carbon supports the nutritional value of plants, while calcium aids in their medicinal benefits and supports their use in traditional diets (Miller et al., 2001; Wroblewitz et al., 2013). Similarly, high nitrogen promotes overall health, and chloride enhances medicinal properties (Grusak and DellaPenna, 1999). Potassium enhances therapeutic uses and, along with calcium, supports both nutritional and medicinal applications (Dorozhkin and Epple, 2002). Silicon and iron further enhance medicinal properties, while a balanced mineral profile bolsters the overall roles of these plants (Martínez-Ballesta et al., 2010).

4. Discussion

4.1. Integrated analysis of the proximate composition and ethno-medicinal profile of plants

The proximate composition analysis of the selected wild edible plant species from various locations in Malakand District reveals significant insights into their nutritional profiles (Table 2).

Regarding moisture content, the plant species showed considerable variation. The highest average moisture contents were observed in Portulaca oleracea L., Rumex dentatus L., and Oxalis corniculata L. High moisture content of these species suggests effective mechanisms for water retention, which could be due to their specific morphological and physiological adaptations (Iqbal et al., 2013).These adaptations might include succulent leaves, extensive root systems, or other water-conserving traits, enabling them to thrive in their respective environments (Abbasi et al., 2015; Ibrahim et al., 2023). Conversely, Chenopodium album L. exhibited the lowest moisture content (Table 2), indicating a potential adaptation to environments where water conservation is crucial. This lower moisture content might reflect a strategy of drought tolerance or efficient water use, allowing it to survive and grow in less favorable conditions (Ibrahim et al., 2023). The high moisture content in Portulaca oleracea, Rumex dentatus, and Oxalis corniculata supports their traditional uses as vegetables and medicinal herbs (Ibrahim et al., 2023). Conversely, the low moisture content in Chenopodium album enhances its therapeutic properties in remedies for piles (Singh et al., 2023).

Moreover, it is crucial to consume foods with a relatively high water content as they play a vital role in various biological processes and prevent dehydration in the body (Lalika et al., 2013). The majority of the wild edible plants examined in this study possess sufficient moisture to fulfill the water needs of human tissues. Our results are consistent with other researchers such as (Hussain et al., 2010; Ullah and Badshah, 2023; Ali et al., 2023) regarding the moisture content of edible wild plants. However, cultivated leafy vegetables exhibit slightly higher moisture content than their wild counterparts (Wallace et al., 2020). In addition, the trend for the moisture contents of the wild edible plants was due to variation in altitudinal zones. According to (Ali et al., 2023), Monotheca buxifolia (Falc.) fruits exhibited a maximum moisture content of 2.08% at lower altitudes, while it was minimal (0.85%) at higher altitudes, which strongly supports our current findings.

The significant variation in dry matter content among the studied plant species (Table 2), underscores diverse adaptive strategies. Chenopodium album, with the highest dry matter content along with Ammaranthus viridis and Medicago polymorpha, suggests these species are well-adapted to conditions where water conservation is essential. High dry matter content indicates a greater resilience to water scarcity and potentially higher nutrient concentration, making these species suitable for environments with limited water availability (Abbasi et al., 2015). Conversely, the low dry matter content of Portulaca oleracea suggests an adaptation for higher water retention, which could be advantageous in maintaining hydration and cellular functions in moist environments. According to (Jedidi et al., 2017), low dry matter and lipid level result in low energy value. The dry matter content of a plant depends on its diverse tissue structure. Moreover, the varied altitudinal ranges contributed to an ascending trend in the dry matter contents (Ali et al., 2023). The findings of the present study align with the results reported by (Kibar and Temel, 2016) and (Tuncturk et al., 2015) for wild edible plants cultivated in Turkey. The elevated dry matter in these plants enhances their effectiveness in therapeutic applications and supports their traditional uses (Street and Prinsloo, 2013).

The ash content, indicative of the mineral content in food material (Anwar et al., 2022), varied significantly among the collected wild edible plants (Table 1). Chenopodium album exhibited the highest ash content, followed by Malva neglecta and Mentha spicata. In contrast, Asparagus officinalis had the lowest ash content. These variations in ash content reflect the differing capacities of these plants to accumulate minerals, which can be influenced by several factors including soil composition, plant physiology, and ecological adaptations (Ibrahim et al., 2023). High ash content in Chenopodium album, Malva neglecta, and Mentha spicata suggests that these species are efficient at mineral uptake and storage, which could be advantageous for growth in mineral-rich soils or for use in nutritionally dense food products (Abdullah et al., 2021). Moreover, the high ash content of Chenopodium album and Malva neglecta signifies their rich mineral content beneficial for therapeutic uses (Yimer et al., 2023). The relatively low ash content in Asparagus officinalis may indicate a different ecological strategy, favoring growth in environments where high mineral accumulation is less critical (Tlahig et al., 2024). The ash content of the wild edible plants was higher than that reported by (Tura et al., 2023) and (Mokria et al., 2022), suggests that the investigated plant species are a rich source of dietary minerals.

Dietary fiber plays a crucial role in promoting the growth and protecting the health of beneficial gut microbiota (Peña-Jorquera et al., 2023). In the current study, Caralluma tuberculata exhibited the highest crude fiber content, followed by Foeniculum vulgare and Melilotus indicus. In contrast, Mentha spicata had the lowest crude fiber content. These variations in crude fiber content highlight the different capabilities of these plants to provide dietary fiber. Caralluma tuberculata, Foeniculum vulgare, and Melilotus indicus, with their high fiber content, could be particularly beneficial for enhancing gut health and supporting the growth of advantageous gut microbiota (Franz et al., 2011; Sun et al., 2021; Plyusnin et al., 2023). This makes them valuable additions to diets aimed at improving digestive health and preventing related disorders. On the other hand, the lower fiber content in Mentha spicata suggests it may have other nutritional benefits but would contribute less to dietary fiber intake compared to the other studied species (Torki et al., 2021). The fiber content of wild edible plants varies depending on the plant species, variety, stage of growth, as well as seasonal and environmental factors (Khanum et al., 2000). In addition, crude fiber content depends on the altitudinal distribution of plants. (Ali et al., 2023) reported high levels of crude fiber in low-lying areas, while lower levels were observed in higher-lying areas. Consuming high-fiber plants helps prevent diseases like diabetes, cancer, heart disease, obesity, high blood pressure, and digestive issues (Anderson et al., 2009). Moreover, the dietary fiber content in Caralluma tuberculata, Foeniculumvulgare, and Melilotus indicus supports their traditional uses in promoting digestive health and weight management (Muhammed et al., 2021).

Lipid content is critically important in nutritional analysis as it serves as a primary energy source (Al Mamun et al., 2023). In the present study, the succulent stem of Caralluma tuberculata had the lowest fat content while the highest mean crude fat content was found in the leaves of Mentha spicata, followed by Mentha longifolia and Rumex hastatus. These results highlight the diverse lipid profiles of the studied plant species, reflecting their potential contributions to dietary fat intake. The high lipid content in Mentha spicata, Mentha longifolia, and Rumex hastatus suggests that these species can be significant sources of dietary fat, which is vital for energy production and various physiological functions (EL-Wahed Ali, 2008; Abd Rashed et al., 2021). Conversely, the lower lipid content in Caralluma tuberculata indicates its suitability for diets requiring lower fat intake (Dutt et al., 2012). The fat content in Mentha spicata, Mentha longifolia, and Rumex hastatus provides energy and nutritional benefits in line with their traditional uses(Talucder et al., 2024). (Seal et al., 2017) reported a high fat content of 6.39±0.03% in the fruits of M. khasianus followed by leaves of S. nigrum (2.01±0.02%) and P. acinosa (1.32 ±0.03%), which is consistent with our study. The variation in the values ​​for crude fat is due to different altitude zones. According to (Ali et al., 2023), the crude fat values in Monotheca buxifolia exhibit an increase from 8.42 ± 0.05 g per 100 g in lower elevation zones to 11.11 ± 1.05 g per 100 g in higher elevation zones. Furthermore, Ziziphus jujuba, Morus alba, and Caralluma tuberculata showed lower crude fat content (2.98±0.2%, 2.02±0.02%, and 2.01±0.03%, respectively), likely due to lipid degradation during drying (Ullah and Badshah, 2023; Ali et al., 2023).

Protein plays an important role in various physiological processes, including growth, body development, tissue repair, enzyme and hormone production, and overall optimal functioning (Farooq et al., 2021). In the present study, Portulaca oleracea was found to have the highest protein content followed by Solanum nigrum while Morus alba showed the lowest protein content (Table 2). Specifically, Portulaca oleracea and Solanum nigrum emerged as notable protein sources in this study, exceeding 6.29 g/100 g. This supports their role as significant dietary sources and validates their traditional uses. Plants with higher protein contents are typically favored in the context of edibility, as they contain essential amino acids that can act as an alternative energy source in cases where carbohydrate metabolism is impaired through glucogenesis (Okerulu and Onyema, 2015). The protein quantity can vary based on factors such as species, climate, edaphology, and other environmental conditions (Adhikari et al., 2022). Compared to previous studies, the wild plants in the Hindukush region demonstrated higher protein content, suggesting that they are rich in protein (Ibrahim et al., 2021; Ullah and Badshah, 2023; Ali et al., 2023). Our findings align more closely with the results of (Ganogpichayagrai and Suksaard, 2020; Sharma et al., 2020). Notably, the crude protein level of these wild species surpasses that of key vegetables like spinach, lettuce, and cabbage. Insufficient protein intake leads to stunted growth, muscle deterioration, edema, abnormal abdominal swelling, and fluid accumulation in the bodies of children (Igile et al., 2013).

Carbohydrates are crucial for human energy metabolism, serving as the primary source of fuel for the body. Analyzing the carbohydrate content in wild edible plants provides insights into their nutritional value and potential dietary contributions (Englyst et al., 2007). The current study found significant variability in carbohydrate levels across different plant species. Ziziphus jujuba exhibited the highest average carbohydrate content (Table 2), making it a substantial source of energy. Asparagus officinalis and Morus alba, with carbohydrate contents of 64.07±0.02% and 62.14±0.3%, indicating their strong potential as energy-rich foods. Portulaca oleracea with lower carbohydrate content of 42.08±0.3% still contributes significantly to the energy intake. In addition, these species differ in the parts used, with Ziziphus jujuba and Morus alba having edible fruits, while Asparagus officinalis and Portulaca oleracea utilize cooked shoots for consumption. The variation in carbohydrate content is influenced by factors such as sunlight, wind and supports their traditional uses as energy sources in traditional diets. Additionally, these environmental factors significantly contribute to the desiccation of fruits, which can have a substantial impact on their carbohydrate contents (Ruiz-Rodríguez et al., 2011). The obtained values indicate that a significant proportion of the wild plants examined in this study possess an adequate carbohydrate content. These findings align with similar results reported by other researchers investigating wild edible plants (Bahadur et al., 2011; Shad et al., 2013; Okerulu and Onyema, 2015; Ullah and Badshah, 2023).

The gross energy value of Solanum nigrum and Bauhinia variegata in the current study indicates their potential as significant sources of dietary energy. This aligns with their traditional uses in cooking and medicinal applications, providing both nutritional and medicinal benefits (Koffi et al., 2020; Anwar et al., 2024). The high energy values in certain species highlight their potential role in enhancing food security and nutrition, particularly in regions where these plants are readily available and consumed (Mehta et al., 2024). (Achaglinkame et al., 2019) also obtained energy values similar to our current study for Adansonia digitate (327–340 Kcal), B. aegyptiaca (266 Kcal), and Tamarindus indica (270–275 Kcal). It has been reported that the energy values (KJ) of plants in lower elevation zones are higher than those in high elevation zones (Ali et al., 2023).

4.2. Looking for association between the variables of proximate composition

The normalized variation matrix (Table 3) might reveal the possibility of a linear association between some parts in a compositional data set (Egozcue et al., 2018). Values below 0.2 suggest proportionality or linear association, whereas values exceeding 1.0 indicate a lack of linear association or proportionality (Boente et al., 2022).

The current study provides a comprehensive analysis of the nutritional contributions of various components in wild edible plants, with an emphasis on the interrelationships among these components. The smallest contributions to energy content from carbohydrates (0.01), moisture content (0.02), dry matter (0.06), fibers (0.06), and energy to moisture content (0.01) suggest a proportional relationship. This indicates that plant species rich in carbohydrates tend to also be high in energy, moisture, dry matter, and crude fibers. The same proportional relationship is observed between energy and moisture content, reinforcing the idea that these nutritional components are interdependent. Similarly, smaller contributions from fibers to moisture content (0.03), ash to moisture content (0.05), and ash to energy content (0.07) imply linear associations among these pairs. These relationships suggest that plant species high in crude fibers and ash content are also rich in energy and moisture content. The largest contributors to variability in the dataset are the logarithmic ratios of fats, proteins, and dry matter relative to other nutrients. Specifically, the significant normalized variation in the logarithmic ratios of fats with dry matter (0.27), fibers (0.23), and proteins with dry matter (0.24) separates these components distinctly in the cluster dendrogram. This differentiation is visually supported by proportional logarithmic ratios of moisture content to carbohydrates (0.02), energy (0.01), and ash content to fibers (0.11), which maintain their respective clusters. The results are further verified through a centered log ratio (CLR) biplot, which explains 66.4% of the total variability in the data. The biplot highlights strong relationships between moisture, energy, and carbohydrates, as well as fibers and ash content, with relative concentration percentages increasing towards the bottom left. Conversely, protein and fat concentrations increase towards the right, while the concentration of dry matter alone increases in the top left of the diagram. The close layout and same ray direction of their vertices suggest a strong interrelationship among these variables, as described by (Pawlowsky-Glahn and Buccianti, 2011). The close proximity of the vertices in the biplot indicates that these variables are associated and tend to vary together. This pattern suggests that increases or decreases in one variable are mirrored by changes in others, likely due to shared properties or interdependencies among carbohydrate, energy, moisture content, and fibers. The strong association between proximate composition variables was further confirmed in the cluster dendrogram (Figure 3) which revealed that closely related variables are grouped into same clusters.

4.3. Mineral content of the wild edible plants.

Wild edible plants contain essential minerals which not only enhances their nutritional value but also underscores their potential to support human health and well-being.

4.3.1. Macro elements

In the current study, Table 4, encompasses both macro and micro elements, revealing significant variations in their concentrations across different species and altitude ranges. The high carbon concentration in plants like Mentha longifolia, Medicago polymorpha, and Bauhinia variegata suggests their significant carbon accumulation capacity, which may be indicative of their physiological and ecological characteristics. (Tognetti et al., 2007; He et al., 2015). Carbon, although not traditionally considered a nutrient, plays a vital role in the production of organic compounds that are essential for human health and well-being (Mie et al., 2017). Plants with higher oxygen concentrations, such as Ziziphus jujuba and Caralluma tuberculata, may exhibit improved nutrient uptake and utilization, leading to increased nutrient levels in their tissues (Farooq et al., 2009; Nile and Park 2014). Foods rich in oxygen may contribute to improved energy production, enhanced oxygenation of tissues, and overall cellular health (Zheng et al., 2003; Nicolle et al., 2004).

Furthermore, the variations in nitrogen and potassium concentrations among the different plant species (Table 4), highlight their potential nutritional benefits. Higher nitrogen fractions in plants like Nasturtium officinale and Asparagus officinalis indicate increased protein content and overall nutritional value, which can be particularly beneficial for individuals following vegetarian diets (Pinela et al., 2017). Likewise, plants with high potassium levels, such as Rumex dentatus, Chenopodium album, and Portulaca oleracea, can help meet the recommended daily intake of this essential mineral and lower the risk of hypertension and cardiovascular diseases (Pohl et al., 2013; Poonia and Upadhayay, 2015; Syed et al., 2016; Khalil et al., 2022). Nitrogen is a fundamental element required for the synthesis of amino acids, proteins, and other essential compounds in living organisms (Zhao et al., 2023). Higher nitrogen fractions in plants may indicate increased protein content and overall nutritional value (Daniel et al., 2022). Plant-based sources with higher nitrogen fractions can be particularly beneficial for individuals following vegetarian or vegan diets, as well as those seeking alternative protein sources (Ahnen et al., 2019).

The significant variation in calcium concentrations in edible wild plants (Table 4) suggests the nutrient diversity they offer. Plants like Bauhinia variegata and Mentha longifolia with high calcium levels can serve as excellent sources of this vital mineral, contributing to meeting the daily calcium intake (Goswami and Ram, 2017; Rehman and Adnan, 2018). Additionally, the analysis of chloride, magnesium, sulphur, phosphorus, and sodium content in these plants underscores their potential contribution to overall mineral intake and their role in supporting various physiological functions such as fluid balance, digestion, muscle function, and bone health (Soetan et al., 2010; Cantwell-Jones et al., 2022). Magnesium is essential for energy production, muscle function, and bone health, while sulphur is crucial for amino acid and protein synthesis (De and De, 2019). Wild edible plants can have higher concentrations of magnesium and sulfur and contribute significantly to overall intake (Tibbetts et al., 2016). Additionally, wild edible plants provide a moderate amount of phosphorus, supporting bone health, energy metabolism, and DNA synthesis (Martínez-Ballesta et al., 2010). Incorporating a variety of wild edible plants with diverse mineral profiles into the diet can help individuals meet their daily nutritional requirements and promote overall health and well-being (Bvenura and Sivakumar, 2017). These findings highlight the importance of exploring and incorporating wild edible plants as a sustainable and nutrient-rich food source, especially in regions with limited access to commercially available fresh produce.

4.3.2. Micro elements

The results of the current study reveal a wide range of silicon, iron, and copper concentrations in various edible wild plant species (Table 4). Silicon, essential for plant growth and defense, ranged from 0.20% to 5.02%, with Oxalis corniculata showing particularly high levels. Plant silicon (Si) is crucial for plant growth, development, and defense against stresses (Khan et al.,, 2021b). Consuming silicon-rich plants like Oxalis corniculata can have beneficial effects on bone strength, immune response, and overall health (Farooq and Dietz, 2015). Iron concentrations varied from 0.21% to 2.03%, with Oxalis corniculata having the highest and Malva neglecta the lowest levels. Iron is an essential micronutrient for humans, playing a crucial role in oxygen transport, energy production, and DNA synthesis (Girerd, 2022). Iron deficiency can lead to anemia, characterized by fatigue, weakness, and impaired cognitive function (Tardy et al., 2020). Consuming foods rich in iron is essential for meeting the recommended daily intake and preventing iron deficiency (Chouraqui, 2022). Incorporating Oxalis corniculata into the diet can contribute to maintaining adequate iron levels as well and promoting overall health, especially in populations vulnerable to iron deficiency (Mishra et al., 2021). However, it is important to note that the iron content alone does not dictate the nutritional value of a food (Fraeye et al., 2020). Malva neglecta may also provide other essential nutrients, vitamins, and minerals that contribute to a balanced diet (Ozturk et al., 2022).

Copper concentrations ranged from 0.40% to 0.99%, with Mentha longifolia and Medicago polymorpha showing lower levels. Understanding copper dynamics in plants is crucial as they are a significant dietary source of this essential mineral (Yruela, 2009). Ensuring adequate intake of copper is necessary to prevent deficiencies and associated health issues (Tahir et al., 2022).. The recommended dietary allowance (RDA) for copper varies based on age, sex, and specific life stages, but for most adults, it ranges from 900 to 1300 micrograms per day (Mustafa and AlSharif, 2018). The results of the current elemental study of selected edible wild plants showed that the studied plant species represent the conspicuous and abundant base of vital and crucial elements. Such knowledge of edible wild plants is of significant importance given the usefulness and potency of these crucial elements in the human body. To promote the conservation and sustainable commercial use of wild edible plants in the region, it is essential to implement sustainable harvesting practices to prevent overexploitation, develop cultivation techniques for high-value plants, and explore diverse commercial applications based on their nutritional benefits (Mokria et al., 2022; Suwardi and Navia, 2023). Integrating local knowledge into conservation strategies, investing in research and development, raising awareness among communities and stakeholders, and establishing regulatory measures are crucial (McKinley et al., 2017). These strategies will help balance ecological preservation with economic viability, ensuring long-term sustainability of these plant resources.

5. Conclusion

The integrated analysis of the proximate composition and ethno-medicinal profile of wild edible plants from various locations in Malakand District revealed significant variation in moisture, dry matter, ash, crude fiber, fat, protein, carbohydrate, and energy content. The high moisture content of certain plant species such as Portulaca oleracea, Rumex dentatus, and Oxalis corniculata supports their freshness and medicinal applications. The high dry matter content of Chenopodium album, Ammaranthus viridis, and Medicago polymorpha enhances their therapeutic properties. Plants with high ash content like Chenopodium album, Malva neglecta, and Mentha spicata exhibit beneficial mineral profiles for medicinal uses. Caralluma tuberculata, Foeniculum vulgare, and Melilotus indicus with high crude fiber content are beneficial for digestive health. Plants like Portulaca oleracea, Solanum nigrum, and Bauhinia variegata with high protein and carbohydrate content provide significant nutritional value and energy. The study also identified associations between different proximate composition variables, highlighting the interconnectedness of these components. The macronutrients and minerals found in the plants further enhance their nutritional and medicinal properties, emphasizing their potential benefits for human health. This comprehensive analysis underscores the nutritional variability and potential dietary contributions of wild edible plants, advocating for their inclusion in diets to enhance food security and nutrition, particularly in regions with limited access to commercially available fresh produce.

Acknowledgements

This Project was funded by the Deanship of Scientific Research (DSR) at King Abdulaziz University, Jeddah, under grant No. (GPIP: 1689-130-2024). The authors, therefore, acknowledge DSR for technical and financial support with thanks.

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Publication Dates

• Publication in this collection
  20 Dec 2024
• Date of issue
  2024

History

• Received
  06 June 2024
• Accepted
  31 July 2024