Analysis of Selected Macro- and Microelement Components in the Indigenous Soybean Cultivars from Regions of the Western Himalaya in India

Abstract

The need to focus on traditional crops for food security has been realized across the globe. ‘Bhat’ is an indigenous soybean variety from the Indian Himalayan Region that has been neglected in recent times due to changing agricultural choices. The present study was conducted on 52 indigenous soybean accessions collected from different villages of Uttarakhand, a hilly state in Northern India, to determine the concentrations of various macro- and microelements in the crop. The analysis used the HHXRF (handheld X-ray fluorescence) spectroscopic technique and atomic absorption spectroscopy (AAS). The findings reveal that the ‘bhat’ is an extremely nutrient-rich crop with calcium being the most significant macronutrient (highest concentration of 27,300 ppm), followed by phosphorus, potassium, and sulphur found in the bhat seeds. The study suggests that samples from Pauri Garhwal (Gwad Khirsu, Bironkhal, Sripur), Tehri (PratapNagar, Singoli), Chamoli (Paini, and Agastyamuni) villages were rich in all the major macronutrients. Among the micronutrients, silicon was found to be the most dominant with the highest concentration of 1920 ppm. This was followed by the dominance of iron, zinc, and manganese in descending order of abundance. Tipri, Chandola Rai Goonth, Chopdiun, and Paini villages were found to have the highest concentrations of microelements. The statistical data analysis confirms that seed samples from different regions are significantly similar in terms of nutrient concentrations, except for Si and Fe, thereby indicating that ‘bhat’ from any of the villages is equally good for consumption. However, seed samples from certain regions were found to be contaminated with heavy metals, indicating soil pollution, which demands attention towards current agricultural practices. The findings show that the crop is an exceptional source of essential and quasi-essential elements and should be promoted to benefit the hill populations.

1. Introduction

Food insecurity continues to be one of the most significant global challenges. While undernutrition in women and children is a global burden [1] its prevalence in mountain communities in different parts of the world has also been established [2,3,4]. Uttarakhand is a hill state in the Western Himalayan region of India that falls into this category. The Garhwal and Kumaon administrative divisions of Uttarakhand have a population of around 3,218,895 and 4,228,998, inhabiting the districts of Chamoli, Dehradun, Haridwar, Pauri Garhwal, Rudraprayag, Tehri, Uttarkashi (in Garhwal), Almora, Bageshwar, Champawat, Nainital, Pithoragarh, and Udam Singh Nagar (in Kumaon) (Figure 1). Published reports have suggested that there is a significant nutritional risk among the rural residents of the hilly areas of the region owing to the challenging mountain terrain, rain-fed agriculture, lack of availability of nutrient-rich food, weak transit linkages, and a lack of awareness of the importance of good nutrition [5,6].

A study on the nutrition status of women in Uttarakhand reported that approximately 43.6% of the total female population were found to be suffering from several levels of chronic energy deficiency (CED) (malnutrition grade I 15.4%, grade II 28.2%) [7]. This could be linked to a lack of essential mineral nutrients in the daily diets of women and children. In particular, calcium and iron deficiency have been associated with increased health risks in female adolescents, pregnant women, and their babies. Similar cases of malnutrition have also been reported in other developing countries. Previous studies have shown that around two billion people worldwide have micronutrient deficiencies, resulting in several health manifestations, and highlight the need to incorporate food with high micronutrients into daily diets [8,9]. An inadequate nutrient intake has also been recognised as one of the immediate causes of undernutrition in children [10].

Consequently, the existing food system, both production and consumption, has come under scrutiny, leading to an increasing focus on incorporating locally farmed nutrient-dense alternate crops. These include crops like millets, soybean, cowpea, indigenous vegetables, etc. [11,12,13].

One such nutritional food from Uttarakhand is the local soybean variety (Glycine max) commonly called ‘Bhat’. ‘Bhat’ is cultivated in hilly regions of Uttarakhand and constitutes an important yet underexploited crop of the region. Available in seeds of varying colour, like black, green, olive, and golden-yellow, and shapes like round, flattened, oval, etc., (Figure 2) the legume has been a traditional food and an integral part of folk remedies of Uttarakhand. The food and medicines prepared from ‘bhat’ have been valued for their nutritional richness and curative properties. The seed husk is also a popular component of animal feed for its “milk-enhancing” properties. According to the Department of Agriculture of the Government of Uttarakhand, ‘bhat’, is one of the most widely grown crops in the area [14], highlighting its popularity in the region despite challenges like mass migration of farmers to cities. However, the crop continues to remain underutilized for its nutritional potential.

Soybean, or the ‘Golden bean’, is widely acknowledged as an outstanding resource for dietary fibre and high-quality protein [15], minerals and elements like calcium, phosphorus, nitrogen, potassium, iron, manganese, and zinc and high protein and amino acid content [16]. Consequently, soybean varieties and their products have been investigated all over the world for their potential as a dietary source and as a therapeutic in the treatment of diabetes, obesity, and digestive, neurological, cardiovascular, and bone ailments [17,18,19]. Soybean is regarded as a vehicle of socio-economic transformation for millions of small and marginal farmers of developing countries. With about 12,200 (1000 MT) production, India is the fifth largest producer of soybeans, contributing immensely towards the national economy through foreign exchange. Different varieties of soybean, including ‘bhat’, are grown in the country that vary greatly in shape, size and colour, the genotypic and phenotypic variations influencing the nutritional diversity.

The age-old ethnomedicinal and therapeutic uses of ‘bhat’ indicate its high nutritive and elemental composition. However, despite the popular traditional use of the crop, there is very little information on the nutritional profile of the cultivars from Uttarakhand. Due to these research gaps, the present study was designed to determine the presence and quantification of several essential and quasi-essential nutrients found in Glycine max ‘bhat’ seeds being cultivated in different districts of Uttarakhand state. These findings will confirm the cultivars with high nutrient contents that can be further selected for sustainable production in the hilly areas, possible value-added product development from the selected superior cultivars, and that will support good nutrition for the population.

2. Methodology

2.1. Sample Collection

Fifty-two distinct soybean ‘Bhat’ samples were collected from different villages of Uttarakhand, falling under different administrative divisions and districts. To avoid any instances of redundancy or repetition, the samples were collected in a randomised fashion from a variety of villages located within each district, preferably at intervals ranging from 5 to 7 kilometres apart. Each year, the crop is harvested by the farmers in October and stored for personal use. These seeds were collected, kept in air-tight bags and stored in clean dry cabinets until further analysis. Although the samples were collected from five districts—namely, Tehri, Pauri, Chamoli, Rudraprayag and Almora—all the samples were arranged in three distinct groups: Tehri, Pauri and Chamoli. This was carried out because the Rudraprayag district is geographically contiguous with Tehri (and was earlier a part of Tehri district, the two regions having been separated only for administrative functioning), while the Pauri and Almora districts share a common geographical boundary and the villages of Almora under study are, location-wise, near the Pauri district (Figure 1). Because of the geographical contiguity of the regions, they share a similar relief and climatic conditions. Tehri, Pauri, and Chamoli are the three major and distinct districts, as the River Ganges and its tributaries form a natural divide between Tehri and Pauri, while Chamoli is characteristically different due to its water resources, altitudes, and agricultural systems. Therefore, for the statistical data analysis, the samples from Rudraprayag district were merged with those from the Tehri, and those from Almora were merged with samples from Pauri districts (

Table 1. Details of soyabean accessions collected from different districts of Uttarakhand state.

Sample No.	Village	Latitude	Longitude	Altitude (in Metres)	District	Seed Coat Colour
1	Agrakhal. New Tehri	30°21	78°32	1749.55	Tehri	Perfect Black
2	New Tehri	30°22	78°26	1021	Tehri	Yellowish White
3	Chamba	30°2	78°23	464.51	Tehri	Perfect Black
4	Dhanolti	30°2	78°14	443.78	Tehri	Yellowish White
5	Pratapnagar	30°25	78°29	371.856	Tehri	Yellowish White
6	Tipri	30°22	78°29	2286	Tehri	Yellow White
7	Singoli	24°96	75°29	371.999	Tehri	Yellow White
8	Badiyargarh	30°17	78°49	1381.00	Tehri	Yellow White
9	Naur	30°13	78°47	371.99	Tehri	Yellow White
10	Ghansyali bhatwara	29°98	78°52	1955.90	Tehri	Yellowish White
11	Bhetti	30°26	78°43	1845.25	Tehri	Yellowish White
12	Ratura	30°30	79°04	861.06	Rudraprayag	Yellowish White
13	Agastyatmuni	30°28	78°98	914.4	Rudraprayag	Yellowish White
14	Dhureli	30°26	79°39	688.84	Rudraprayag	Light Black
15	Pauri	30°90	78°45	1765.09	Pauri	Yellowish White
16	Chandola Rai Goonth	30°12	78°7	371.99	Pauri	Yellowish White
17	Falswari	30°14	78°69	345.00	Pauri	Perfect Black
18	Gwad Khirsu	30°10	78°52	1699.565	Pauri	Yellowish White
19	Chopdiun	30°11	78°85	1635.55	Pauri	Yellowish White
20	Bironkhal	29°50	79°1	1505.10	Pauri	Yellowish White
21	Toli	30°10	78°47	345.00	Pauri	Yellowish White
22	Kalkhobiya	29°45	79°4	1332.89	Pauri	Perfect Black
23	Markhola	29°82	78°96	1131.89	Pauri	Greenish Yellow
24	Santudhar	30°0	78°49	345.00	Pauri	Yellowish White
25	Satpuli	29°53	78°43	1819.656	Pauri	Yellowish White
26	Pokhra	29°54	78°55	1111.91	Pauri	Yellowish White
27	Kota	29°46	78°5	345.99	Pauri	Yellowish White
28	Jaiharikhal	29°51	78°4	1570.939	Pauri	Perfect Black
29	Dugadda	29°48	78°36	745.84	Pauri	Yellowish White
30	Maral	30°75	78°6	355.00	Pauri	Yellowish White
31	Sripur	30°18	78°78	1765.99	Pauri	Yellowish White
32	Barsori	30°14	78°8	371.99	Pauri	Yellowish White
33	Mandakhal	30°11	78°81	1171.956	Pauri	Yellowish White
34	Gawad	30°17	78°85	1733.99	Pauri	Yellowish White
35	Sankhyakot	30°40	79°31	1299.99	Almora	Black
36	Chaumu	30°44	79°31	1289.3	Almora	Black
37	Sironiyan	30°35	79°31	1297.4	Almora	Black
38	Sirtoli	30°29	79°28	345.00	Chamoli	Greenish yellow
39	Kaleshwar	30°28	79°24	369.00	Chamoli	Yellowish White
40	Chamola	30°20	79°24	765.048	Chamoli	Greenish Yellow
41	Tauli Daduli	30°24	79°23	369.00	Chamoli	Light Black
42	Uttaraun	30°31	79°30	859.536	Chamoli	Yellowish White
43	Kafalkhet	30°35	79°31	1392.02	Chamoli	Yellowish White
44	Kathur	30°41	79°30	1052.474	Chamoli	Yellowish White
45	Pakhi	30°46	79°44	1716.634	Chamoli	Light Black
46	Dwing	30°48	79°47	1087.22	Chamoli	Greenish Yellow
47	Paini	30°53	79°51	1874.52	Chamoli	Greenish Yellow
48	Gulabkoti	30°50	79°49	2044.903	Chamoli	Perfect Black
49	Joshimath	30°46	79°43	2042.465	Chamoli	Light Black
50	Dungari	30°14	79°37	369.00	Chamoli	Perfect Black
51	Gudam State	30°03	79°52	345.00	Chamoli	Yellowish White
52	Lolti	30°04	79°51	345.00	Chamoli	Light Black

).

2.2. Sample Preparations

The samples were further selected based on their moisture content. The oven-drying method was used to analyse the moisture content in which the samples were oven-dried for three to four hours and weighed before and after drying to determine the moisture content in the seeds. The samples with a moisture content of 15% or less were selected for further processing. The selected samples were sun-dried for approximately two to three days, followed by drying in a hot air oven for another two to three days at a temperature of 50 °C, with relative humidity being less than 12%. The oven-dried samples were finely crushed into powder form and used for further analysis.

2.3. HHXRF Analysis

HHXRF is an analytical technique for the atomic emission spectroscopy-based qualitative and quantitative evaluation of biological samples. It is a portable, easy-to-handle, non-destructive device for the estimation of all types of macro- and microelements and heavy metals in the sample. For an initial evaluation, a set of 5 samples (New Tehri, Chamba, Bironkhal, Gawad Khirsu, and Chamola villages), were analysed for mineral composition using HHXRF technique. The powdered samples were subjected to elemental analysis using a handheld X-ray fluorescence (HHXRF) spectrometer in the Nuclear Physics Division, Bhabha Atomic Research Centre (BARC), Mumbai, India. The powdered samples were placed in a cubic box and irradiated by a rhodium X-ray tube with high-voltage power of up to 50 volts. The samples were irradiated with primary X-rays to excite the elements present in the sample, which further produces secondary fluorescent X-rays specific for each element with characteristic energy. Measuring these energies could determine qualitatively and quantitatively the different types of elements present. The results indicated the presence of high levels of minerals such as Ca, Si, Fe, Zn in all the analysed samples. Subsequently, all the samples were subjected to spectroscopic analysis for a detailed nutrient evaluation.

2.4. Atomic Absorption Spectroscopy

All 52 samples were further analysed on atomic absorbance spectroscopy (AAS) Agilent and 240 FS (2023). The AAS technique is frequently used to quantify metallic and non-metallic elements present in any biological fluid, organic or inorganic solution and tissue sample [20]. The analyte in the solution is exposed to heat energy and combustible gases to form free ions that further become excited to a higher energy state after absorbing a specific wavelength passed through the flame. The amount of light absorbed to elevate the metal atoms to an excited state is measured to determine the concentration of the element using a calibration curve. A higher absorbance depicts the higher concentration of the element present in the solution (Beer Lambert’s law) [21,22].

For sample preparation, 0.5 g of the powdered sample was weighed and suspended in 70% (v/v) nitric acid for digestion. The sample was then placed on a hot plate for further digestion till a colourless solution was obtained. The final volume was made up to 100 mL with 2% (v/v) nitric acid. The solution was filtered using Whatman filter paper, and the filtrate was further processed for atomization followed by absorption under a high-temperature flame. Before sample analysis, the plate reader was calibrated with solutions of known concentration. Thereafter, the concentrations of the minerals in the test solution were analysed using calibration curves and displayed on the screen along with the spectrum showing peaks obtained during the analysis.

2.5. Statistical Analysis

All the experiments were performed in triplicate. The data on concentrations of different elements in all the samples were tabulated in three groups (Tehri, Pauri and Chamoli). The location, namely Tehri, Pauri and Chamoli, subsumes the variations in the soil, temperature, precipitation and other factors and serves as a single factor. The data were analysed using ANOVA of completely randomized design (CRD) in SPSS software version 29.0. The mean values were determined and the significance level was determined at p ≤ 0.05.

3. Results

‘Bhat’ is a popular legume of the hill state Uttarakhand in India valued for its several health benefits. However, scientific evaluation of indigenous ‘bhat’ cultivars is scarce. The present study investigates the nutrient profile of ‘bhat’ accessions collected from local growers in hilly areas of Uttarakhand, India to establish this crop as a valuable and sustainable food choice. The study shows that the samples collected from different locations, varying in altitudes from 369 to 2200 m, exhibited variations in shape, size and seed coat colour. All the samples were collected from local farmers from varied locations where the agricultural practices are different, suiting local inputs availability and conditions. The dynamic environmental factors like weather conditions, temperature, pre-sowing rains, biotic and abiotic stresses influence the cultivar quality. However, most of the seeds are locally preserved, and untreated in most cases. The analysis revealed that all the ‘bhat’ samples showed high amounts of Ca, ranging from 3086 ppm to 27,300 ppm, the highest amounts being registered in samples from Agrakhal, New Tehri (

Table 2. Macronutrient (Ca, P, K, S) composition in the ‘bhat’ samples from Uttarakhand.

Tehri
		Calcium	Phosphorus	Potassium	Sulphur
Sno	Villages	Mean Value (ppm) + SD	Mean Value (ppm) + SD	Mean Value (ppm) + SD	Mean Value (ppm) + SD
1	Agrakhal. New Tehri	27,300 ± 0.3	0.66 ± 1.1	0 ± 0	0 ± 0
2	New Tehri	23,023 ± 0.5	0.33 ± 0.5	0 ± 0	0 ± 0
3	Chamba	25,440 ± 0.6	0.33 ± 0.5	0 ± 0	0 ± 0
4	Dhanolti	22,038.67 ± 0.5	1 ± 1	0 ± 0	0 ± 0
5	Pratapnagar	10,416.67 ± 1.2	232.66 ± 2.5	170 ± 0.1	0 ± 0
6	Tipri	24,989 ± 0.9	1 ± 1.7	0 ± 0	0 ± 0
7	Singoli	11,536 ± 0.7	0.33 ± 0.5	288 ± 1	0 ± 0
8	Badiyargarh	7460 ± 0.1	0.33 ± 0.5	197.33 ± 0.5	0 ± 0
9	Naur	9053 ± 1.6	193.33 ± 6.6	0 ± 0	0 ± 0
10	Ghansyali bhatwara	23,312.617 ± 0.9	0.33 ± 0.5	0 ± 0	0 ± 0
11	Bhetti	10,802 ± 1.2	0.66 ± 1.1	0 ± 0	0 ± 0
12	Ratura (Rudraprayag)	15,584 ± 0.4	0 ± 0	251.66 ± 0.8	2.886751 ± 3
13	Agastyatmuni (Rudraprayag)	13,320.67 ± 0.1	0.333 ± 0.5	282.66 ± 1.6	160 ± 0
14	Dhureli (Rudraprayag)	8343 ± 2.6	249.66 ± 0.5	0 ± 0	168 ± 2.6
15	Pauri	14,705 ± 0.5	0 ± 0	0 ± 0	209.33 ± 0.006
16	Chandola Rai Goonth	19,305 ± 01.5	0 ± 0	0 ± 0	0 ± 0
17	Falswari	4694.333 ± 0.5	0 ± 0	191 ± 0	160 ± 1
18	Gwad Khirsu	7454 ± 2.5	139 ± 1.2	296 ± 0	262.66 ± 0.7
19	Chopdiun	11,051.33 ± 3.2	0 ± 0	140 ± 0	0 ± 0
20	Bironkhal	7800 ± 0.05	241.66 ± 2.09	480 ± 0	269.66 ± 0.6
21	Toli	10,377.67 ± 3.2	0 ± 0	170 ± 0	170 ± 0.5
22	Kalkhobiya	13,922.67 ± 1.3	0 ± 0	0 ± 0	0 ± 0
23	Markhola	20,303.33 ± 0.03	0 ± 0	0 ± 0	189.66 ± 0.4
24	Santudhar	22,810 ± 0.005	0 ± 0	0 ± 0	0 ± 0
25	Satpuli	20,343.333 ± 1.7	0 ± 0	0 ± 0	0 ± 0
26	Pokhra	9600 ± 1.5	0 ± 0	133 ± 0	190 ± 1
27	Kota	17,200.33 ± 0.5	0 ± 0	0 ± 0	230 ± 0.8
28	Jaiharikhal	20,382.33 ± 2.9	0 ± 0	0 ± 0	0 ± 0
29	Dugadda	15,400.67 ± 1.1	0 ± 0	186 ± 0	331 ± 0.7
30	Maral	21,800.33 ± 0.5	0 ± 0	150 ± 0	0 ± 0
31	Sripur	12,801 ± 0.7	63.33 ± 1.09	475 ± 0	351.66 ± 0.3
32	Barsori	3535 ± 1.6	253.3 ± 2	330 ± 0	0 ± 0
33	Mandakhal	5293.667 ± 1.7	0 ± 0	430 ± 0	310.33 ± 0.5
34	Gawad	18,651.33 ± 3.2	0 ± 0	0 ± 0	0 ± 0
35	Sankhyakot (Almora)	3411.667 ± 2.8	180.333 ± 1.5	190.66 ± 1.1	0 ± 0
36	Chaumu (Almora)	2178 ± 2.6	0 ± 0	17700 ± 0	3250 ± 2.6
37	Sironiyan (Almora)	3086.333 ± 1.5	5830 ± 1	17700 ± 0	2941.33 ± 2.3
Chamoli
38	Sirtoli	18,910 ± 0.5	0 ± 0	0 ± 0	481.33 ± 1
39	Kaleshwar	12,606 ± 0.6	0 ± 0	0 ± 0	0 ± 0
40	Chamola	7370 ± 0.5	0 ± 0	193 ± 0	190 ± 0.005
41	Tauli Daduli	12,800.33 ± 2.5	0 ± 0	0 ± 0	0 ± 0
42	Uttaraun	19,031.33 ± 3.2	0 ± 0	0 ± 0	0 ± 0
43	Kafalkhet	2745.333 ± 1.1	179.66 ± 0.5	270.66 ± 1.1	120.66 ± 2.2
44	Kathur	4389.667 ± 1.5	0 ± 0	620.66 ± 1.1	719.66 ± 1.2
45	Pakhi	18,051.33 ± 2.3	0 ± 0	170 ± 0	450.66 ± 1.5
46	Dwing	12,741.33 ± 2.2	0 ± 0	250 ± 0	200.33 ± 0.9
47	Paini	6591 ± 2.6	231 ± 1.7	170 ± 0	200.66 ± 0.4
48	Gulabkoti	11,341.33 ± 1.5	0 ± 0	0 ± 0	151.33 ± 0.6
49	Joshimath	17,992.67 ± 0.5	0 ± 0	0 ± 0	0 ± 0
50	Dungari	19,342.67 ± 2.5	0 ± 0	0 ± 0	0 ± 0
51	Gudam State	4981.33 ± 1.5	0 ± 0	150.33 ± 0.5	139.66 ± 0
52	Lolti	9840 ± 0.1	0 ± 0	109.66 ± 0.5	0 ± 0
	Significant difference	NS	NS	NS	NS
	LSD	0.05	0.05	0.05	0.05

NS—Non significant difference; LSD—Least significant difference.

, Figure 3). Moreover, phosphorus (0–5830 ppm), potassium (0–17,700 ppm) and sulphur (0–3250 ppm) were also recorded as the most pronounced essential nutrients in ‘bhat’ seeds.

The ‘bhat’ samples under study were also found to be enriched with zinc, iron, manganese, and majorly with silicon (

Table 3. Microelements such as zinc, silicon, iron and manganese present in the samples from Uttarakhand.

Tehri
		Zinc	Silicon	Iron	Manganese
S. No.	Village	Mean Value (ppm) + SD	Mean Value (ppm) + SD	Mean Value (ppm) + SD	Mean Value (ppm) + SD
1	Agrakhal. New Tehri	8 ± 1	0 ± 0	190 ± 1	1.33 ± 1.5
2	New Tehri	8 ± 1	903 ± 5.1	208 ± 4	0 ± 0
3	Chamba	8 ± 1	1430 ± 1	193 ± 6	0 ± 0
4	Dhanolti	7 ± 1	1380 ± 2	138 ± 4	0.66 ± 0.57
5	Pratapnagar	15 ± 1.2	961 ± 5	105 ± 5	0 ± 0
6	Tipri	7 ± 1.2	1606 ± 1.7	126.66 ± 1.1	67 ± 1
7	Singoli	12 ± 0.1	990 ± 4	145 ± 5	0 ± 0
8	Badiyargarh	25 ± 0.3	910 ± 1	117 ± 0	0.1 ± 0.1
9	Naur	14 ± 0.2	1350 ± 5	106.66 ± 0.5	0 ± 0
10	Ghansyali bhatwara	10 ± 0.4	1030.333 ± 0.5	180 ± 0	0.1 ± 0.1
11	Bhetti	14 ± 0.1	931.33 ± 3.2	75 ± 2	0 ± 0
12	Ratura (Rudraprayag)	8 ± 1	1620 ± 0	104 ± 0	0 ± 0
13	Agastyatmuni (Rudraprayag)	12 ± 1	1640 ± 1.3	94 ± 1.4	0 ± 0
14	Dhureli (Rudraprayag)	10 ± 1	1430 ± 1.4	55 ± 1.4	0 ± 0
15	Pauri	8 ± 0.8	1250 ± 2	75 ± 2	0 ± 0
16	Chandola Rai Goonth	10 ± 1.6	1650 ± 2	107 ± 2	40.33 ± 0.05
17	Falswari	23 ± 0.3	1300 ± 1	47 ± 1.7	0 ± 0
18	Gwad Khirsu	17 ± 0.1	1360 ± 2	69 ± 2	0.2 ± 0.2
19	Chopdiun	15 ± 0.1	1598 ± 2	73 ± 1.4	0.06 ± 0.5
20	Bironkhal	20 ± 0.1	1530 ± 1	94 ± 1.5	0 ± 0
21	Toli	9 ± 0.6	1563 ± 0.06	72 ± 0.3	0 ± 0
22	Kalkhobiya	9 ± 2.4	1785 ± 0.3	82 ± 0.3	0 ± 0
23	Markhola	7 ± 0.5	1850 ± 1	87 ± 1	0 ± 0
24	Santudhar	6 ± 1.2	1370 ± 0.7	111 ± 1	0 ± 0
25	Satpuli	6 ± 1.2	1280 ± 1	90 ± 1	0 ± 0
26	Pokhra	15 ± 2	1330 ± 1	71 ± 2	0 ± 0
27	Kota	6 ± 1	1071 ± 1	93 ± 1	0 ± 0
28	Jaiharikhal	8 ± 0	1540 ± 0	104.33 ± 1.08	0 ± 0
29	Dugadda	11 ± 2	1271 ± 1	83 ± 1	0 ± 0
30	Maral	7 ± 2	1332 ± 2	121 ± 1	0 ± 0
31	Sripur	12 ± 3	1170 ± 1	125 ± 0	0 ± 0
32	Barsori	24 ± 3.6	1390 ± 1.7	70 ± 1	0 ± 0
33	Mandakhal	22 ± 0	1291 ± 1.6	80 ± 0	0 ± 0
34	Gawad	9 ± 2	1792 ± 0.7	89 ± 1	0 ± 0
35	Sankhyakot (Almora)	22 ± 1.2	1320 ± 2	61 ± 0.6	0 ± 0
36	Chaumu (Almora)	62 ± 2	1010 ± 0.2	87 ± 2	47 ± 1
37	Sironiyan (Almora)	55.34 ± 1	880 ± 0.5	119 ± 0	34 ± 0
Chamoli
38	Sirtoli	12.66 ± 1.05	1890 ± 1	116 ± 2	0 ± 0
39	Kaleshwar	7 ± 0.01	1500 ± 2.6	57 ± 2	0 ± 0
40	Chamola	18 ± 1.2	1230 ± 1.6	73 ± 2.2	0 ± 0
41	Tauli Daduli	0 ± 0	1520 ± 1.5	70 ± 0	36 ± 0
42	Uttaraun	7 ± 0.4	1920 ± 1.3	116 ± 2.1	0 ± 0
43	Kafalkhet	24 ± 2	1140 ± 0.5	54 ± 3	0 ± 0
44	Kathur	24 ± 2	980 ± 0.4	80 ± 1	0 ± 0
45	Pakhi	0 ± 0	1430 ± 0.6	91.66 ± 0.08	0 ± 0
46	Dwing	12 ± 0.1	1677 ± 1.1	80 ± 1.7	0 ± 0
47	Paini	13 ± 0	1550 ± 4	65 ± 2.3	45 ± 0
48	Gulabkoti	10 ± 1	1150 ± 1	67 ± 3	0 ± 0
49	Joshimath	9 ± 1	1900 ± 0.8	101 ± 0.4	0 ± 0
50	Dungari	7 ± 1	1571 ± 0.6	103 ± 3	0 ± 0
51	Gudam State	23 ± 1.6	1680 ± 0.3	75 ± 2.6	0 ± 0
52	Lolti	13 ± 1	1120 ± 1	81 ± 0.56	0 ± 0
	Significant difference	NS	***	***	NS
	LSD	0.05	0.05	0.05	0.05

p value ≤ 0.05 indicates the groups showing significant difference. NS denotes non significant difference (samples are not significantly different); *** denotes significant difference.

). The amounts of Si found in different villages were significantly different from each other, with the highest amount being 1920 ppm, and the next highest concentration of 1900 ppm being recorded in samples from Chamoli. The iron content in different samples was also significantly different, with the highest concentration of 208 ppm followed by 193 ppm being present in the samples from Narendranagar and Chamba villages in the Tehri district.

The analysis also revealed the presence of toxic elements in the ‘bhat’ samples from some areas (

Table 4. Toxic elements such as cadmium, thorium, aluminium and strontium present in the samples from Uttarakhand.

Tehri
		Cadmium	Thorium	Aluminium	Strontium
S. No.	Village	Mean Value (ppm) + SD	Mean Value (ppm) + SD	Mean Value (ppm) + SD	Mean Value (ppm) + SD
1	Agrakhal. New Tehri	0 ± 0	50 ± 4.5	0 ± 0	10 ± 1
2	New Tehri Narendranagar	31.66 ± 1.52	44.33 ± 2.08	0 ± 0	9 ± 1
3	Chamba	28 ± 2.6	50 ± 0	0 ± 0	23 ± 0
4	Dhanolti	50.33 ± 0.57	54.332.3 ±	0 ± 0	13 ± 0.2
5	Pratapnagar	43.33 ± 0.57	32.66 ± 2.5	1800 ± 0.9	9.66 ± 1.2
6	Tipri	0 ± 0	65.33 ± 3.0	0 ± 0	29.66 ± 0.8
7	Singoli	36.66 ± 0	41.66 ± 4.5	2875.66 ± 1.1	19 ± 1
8	Badiyargarh	22.33 ± 0.57	40.33 ± 1.5	2003.66 ± 8	6 ± 1
9	Naur	0 ± 0	39.66 ± 2.5	2304.66 ± 6	20.66 ± 2
10	Ghansyali bhatwara	0 ± 0	55.66 ± 1.5	0 ± 0	15 ± 2
11	Bhetti	25 ± 0	34.33 ± 2.5	0 ± 0	15.33 ± 1
12	Ratura (Rudraprayag)	32.66 ± 2.5	54.33 ± 1.5	0 ± 0	15 ± 0
13	Agastyatmuni (Rudraprayag)	0 ± 0	48 ± 1.3	0 ± 0	13 ± 1
14	Dhureli (Rudraprayag)	27 ± 2.6	38 ± 2	2700.33 ± 2.5	12 ± 2
15	Pauri	28 ± 7	65 ± 0	0 ± 0	17 ± 1
16	Chandola Rai Goonth	0 ± 0	66 ± 3	0 ± 0	25 ± 2
17	Falswari	0.33 ± 0.5	33 ± 3.5	1806 ± 4	6.66 ± 1
18	Gwad Khirsu	32.66 ± 0.5	36.33 ± 2.5	2302 ± 2	6 ± 0
19	Chopdiun	0 ± 0	49.33 ± 0.5	0 ± 0	7.66 ± 0.5
20	Bironkhal	28.33 ± 0.5	52.33 ± 2.5	2199.66 ± 0.5	13.66 ± 1
21	Toli	0 ± 0	45.33 ± 0.5	300.66 ± 2	21.66 ± 0.2
22	Kalkhobiya	34 ± 1	54 ± 1.3	1801.33 ± 3.2	30 ± 0
23	Markhola	0 ± 0	57 ± 1	0 ± 0	11.33 ± 0.5
24	Santudhar	32 ± 0	60.66 ± 0.5	0 ± 0	15.33 ± 0.5
25	Satpuli	0 ± 0	53 ± 1	0 ± 0	11.66 ± 0.5
26	Pokhra	35 ± 0	43.33 ± 1.5	1601.66 ± 0.37	9 ± 0
27	Kota	35.33 ± 0.5	47.66 ± 1.1	0 ± 0	24 ± 2
28	Jaiharikhal	0 ± 0	59.66 ± 2	0 ± 0	10 ± 0
29	Dugadda	31.33 ± 1.1	45 ± 1	1600.33 ± 1.5	9.33 ± 1
30	Maral	0 ± 0	48 ± 1.3	0 ± 0	13 ± 2.6
31	Sripur	27 ± 2	40 ± 1.2	0 ± 0	7.33 ± 1.5
32	Barsori	41.33 ± 1.2	30 ± 1	1900.66 ± 1.1	7 ± 0
33	Mandakhal	39.33 ± 0.004	52 ± 1	2400 ± 0	12 ± 1
34	Gawad	0 ± 0	71 ± 1	0 ± 0	50.66 ± 1.1
35	Sankhyakot (Almora)	29.66 ± 1.5	29.33333 ± 3.2	1800.33 ± 0.5	8 ± 1
36	Chaumu (Almora)	28.33 ± 1.5	0 ± 0	0 ± 0	7 ± 0.3
37	Sironiyan (Almora)	28 ± 1.7	0 ± 0	1300 ± 0.06	12 ± 0
Chamoli
38	Sirtoli	0 ± 0	70.66 ± 2	0 ± 0	14 ± 1
39	Kaleshwar	0 ± 0	40.33 ± 1.5	0 ± 0	14.66 ± 3.5
40	Chamola	33.33 ± 1.5	38.33 ± 2	2301 ± 1.7	8.66 ± 1.1
41	Tauli Daduli	0 ± 0	54 ± 3	0 ± 0	11 ± 1
42	Uttaraun	0 ± 0	57 ± 2	0 ± 0	14.5 ± 2
43	Kafalkhet	22.66 ± 1.5	27 ± 2.3	1900 ± 5	5 ± 1
44	Kathur	26.33 ± 0.57	35 ± 1	2700 ± 2	10 ± 1
45	Pakhi	0 ± 0	58.33 ± 1.5	0 ± 0	12.66 ± 2
46	Dwing	0 ± 0	58.66 ± 0.5	0 ± 0	10.33 ± 1.5
47	Paini	36.33 ± 3.5	36.66 ± 2	0 ± 0	7.33 ± 1.5
48	Gulabkoti	30.33 ± 2.5	46 ± 2.3	2001 ± 2.6	14 ± 1
49	Joshimath	42 ± 2.6	62 ± 2	0 ± 0	15 ± 1
50	Dungari	34.66 ± 5	54.66 ± 2.5	0 ± 0	15.66 ± 2.08
51	Gudam State	0 ± 0	29.33 ± 1.1	2201 ± 1.7	7.66 ± 1.5
52	Lolti	0 ± 0	42.66 ± 0.5	0 ± 0	11.33 ± 0.5
	Significant difference	NS	NS	NS	NS
	LSD	0.05	0.05	0.05	0.05

NS—non significant difference.

). Trace amounts of toxic elements such as aluminium (0–2875 ppm), strontium (5–50 ppm), thorium (0–71 ppm) and cadmium (0–50 ppm) were seen in some samples, with Sr being present in almost all the samples. The sample with the highest Al content was reported to be from Singoli village (Figure 4). It was observed that samples from Tehri villages had more pronounced concentrations of heavy metals like Al and Cd, while those from Pauri Garhwal had low yet accountable concentrations of Th and Sr.

The statistical data analysis revealed that, except for Si and Fe, there was no significant difference in the amounts of nutrients within or among the districts. This indicates that ‘bhat’ from any part of Uttarakhand under study is good enough for consumption. However, samples from Pauri Garhwal (Gwad Khirsu, Bironkhal, Sripur, Sironiyan), Tehri (Agastyamuni), Chamoli (Kafalkhet and Paini) had all major macronutrients, Ca, P, S, and K, while samples from Tipri, Chandola Rai Goonth, Chopdiun and Paini are best for the microelement. After comparing all the samples, it was determined that Gwad Khirsu, Sironiyan and Paini exhibited the highest number of both macro- and microelements. The samples from Chopdiun, Maral, Chaumu (Pauri), Lolti and Kaleshwar (Chamoli) were found to have the least amount of toxic heavy metals.

To summarize, the results show that the indigenous soybean variety ‘bhat’ from the hills of Uttarakhand is enriched with a high content of essential elements and can cater to the nutrient requirements of the residents. Among the macronutrients, Ca is the most abundant element present in all the accessions, followed by P in most of the samples and K and S in some of the samples collected. Analysis for micronutrients showed that Si was the most pronounced element, followed by substantial amounts of Fe and Zn in all the seed samples, and manganese in some of the samples. However, the presence of heavy metals like Cd and Al in some of the accessions, though within tolerable intake levels, poses a challenge to safe consumption if not paid attention.

4. Discussion

The results show that the native crop of Uttarakhand‘bhat’is enriched with a high content of essential macro- and microelements. The presence of macroelements like P, K, and S was confirmed in most of the samples while high amounts of Ca were found in all 52 samples collected from different villages in Uttarakhand. Previously, Croatian soybeans have been reported to have about 1.9 mg of calcium per gm sample [23] while soybeans from Nigeria were evaluated and found to contain around 3 mg Ca per gram of the seed sample [24]. In our study, a range of about 0.3 mg/gm to 27 mg/gm calcium in ‘bhat’ samples indicates it to be a Ca reservoir that can be a potential solution to the problems of calcium deficiency among the residents of hilly areas. At high altitudes, the milk productivity of animals is reportedly very low, contributing only 1.15% to dairy [25], leading to a shortage of dairy food and subsequent calcium deficiency among the inhabitants, particularly the female population. In this scenario, incorporation of ‘bhat’ into the regular diet can be particularly useful for the women in these regions. Calcium is also one of the most essential elements which can help in decreasing the risk of osteomalacia in children by rapidly improving bone mineralization; it also helps in muscle contraction and maintaining healthy heart function [26]. Consequently, ‘bhat’ can serve as nutritious food and could be an inexpensive way to prevent calcium deficiency and bone-related health problems prevalent among the residents of the hills.

Quasi-elements are required in comparatively lower amounts yet essentially needed for the proper functioning of the human body. Good amounts of the microelements Si, Fe, Zn, and Mn were also confirmed in the ‘bhat’ samples, indicating their suitability for improving bone health, and boosting haemoglobin, overall immunity, and biochemical functioning when used as a dietary intake. Silicon plays a crucial role in the biochemistry of subcellular enzymatic reactions [27]. It is also important for strengthening bones, improving the immune response, and neuronal and connective tissue health [28,29,30,31]. Besides its role in bone calcification, connective tissue matrix formation and prevention of the ageing process have also been reported [32]. While Fe is essential for blood haemoglobin regulation, Zn and Mn participate in various biological processes, such as ROS detoxification, enzyme synthesis, hormone regulation, physical development, regulation of the immune system, circulatory system and reproductive system [33,34]. Comparing the number of elements required for daily consumption as per RDI suggested by the National Institute of Health (NIH 2023) [35], this crop can prove to be an essential source of nutrients to maintain the body’s metabolism and healthy development. The selected ‘bhat’ samples can be further earmarked and promoted for cultivation or product development.

One important observation in this study was the detection of heavy metals like cadmium (Cd) and aluminium (Al) in some seed samples of ‘bhat’, indicating potential contamination of soil in specific areas of Uttarakhand hills and the heavy metal phytoextraction potential of ‘bhat’ cultivars. As per a report [36], the recommended tolerable monthly intake for Al, Cd and Sr are 2000 µg, 25 µg and 400 µg, respectively. According to this information, the consumption of ‘bhat’ in cuisine can be considered safe. These observations could be linked to increased unscientific anthropogenic activities like unplanned pesticide usage, increased irresponsible tourist activities, etc., in the hilly areas, accounting for elevated heavy metals in the hilly soils. Soybeans are known to have the potential for phytoremediation and phytoextraction, including metals like cadmium and lead [37,38,39]. The present study confirms the phytoextraction property of ‘bhat’ and sends out an alert that increased soil pollution can contaminate the food crop.

Overall, the findings of the study can be used for ‘bhat’ prioritization, propagation, consumption and the formulation of soybean nutrient management strategies in the region.

5. Conclusions

The present study confirms that ‘bhat’ seeds from different villages of Uttarakhand, India, contain good quantities of macro- (Ca, P, S, K) and microelements (Fe, Zn, Si, Mn). The samples are rich in calcium, as high as 27,300 ppm, silicon (highest concentration being 1920 ppm) and iron (highest concentration being 208 ppm). Statistical analysis reveals that ’bhat’ seeds from any of the locations under study are significantly similar in elemental richness, irrespective of their origin and method of cultivation. However, the presence of heavy metals like Cd and Al in some seed samples indicates increasing soil contamination in the hills of Uttarakhand, India, and further signifies the need for awareness and implementation of good agricultural practices. Further, the nutritional superiority of the indigenous soybean must be utilized for product development and further promotion in national and international markets.