Introduction

Plants serve as valuable repositories of medicinal compounds and play a significant role in the lives of ethnic and tribal communities, particularly in remote and rural areas. Globally, these plants have long been used to treat a wide range of human and animal ailments1. Their role in traditional healthcare systems has attracted increasing scientific interest, especially for the discovery of bioactive compounds with pharmacological potential. In addition to therapeutic properties, many medicinal plants also offer important nutritional benefits, containing essential macronutrients like carbohydrates, proteins, and fats that support various physiological and metabolic functions in the human body1,2. Plants are rich in phytochemicals such as alkaloids, flavonoids, phenols, saponins, and tannins, many of which exhibit antioxidant, antimicrobial, and anti-inflammatory properties3. These secondary metabolites are often responsible for the therapeutic effects observed in traditional medicine. The chemical diversity of these bioactive compounds has led to the development of several modern drugs, either directly from plant extracts or through synthetic modification4. Owing to their accessibility, affordability, and lower risk of side effects compared to synthetic drugs, plant-based medicines remain widely used, especially in rural regions5.

According to the World Health Organization, approximately 80% of the global population depends on traditional plant-based medicines for their primary healthcare needs. This reliance highlights the importance of preserving both the plant species and the associated indigenous knowledge6. In regions like the Western Himalayas, medicinal plants are not only used for healthcare but also contribute to food security, cultural practices, and livelihood. However, increased modernization and environmental degradation pose threats to this traditional knowledge and the biodiversity that supports it5.

Plants fulfill fundamental needs for human existence, providing necessities like food, fiber, shelter, and medicinal remedies. Particularly in remote areas, traditional herbal remedies are favored for healthcare. Even in regions where conventional medicine is prevalent, there’s a growing interest in plant-based natural remedies1. Medicinal plants contain diverse physiologically functional compounds, including nutrients and phytoconstituents, which exert various physiological responses on humans. They serve as reservoirs of chemical compounds like alkaloids, flavonoids, and saponins2. The biologically active components found in medicinal herbs are utilized for preventive purposes and for treating various infectious diseases. Due to their medicinal properties, these plants have been utilized across a wide geographical area. Folklore medicines have been employed to treat numerous ailments such as malaria, epilepsy, diarrhea, dysentery, as well as fungal and bacterial infections. Secondary metabolites are synthesized through various pathways in different plant species3. Deficiencies in micronutrients and minerals within the diet can result in long-term detrimental impacts on human health, potentially leading to nutrient deficiency disorders4. Historically, therapeutic remedies were exclusively sourced from herbs, whether they manifest as basic parts of plants or complex mixtures like crude extracts5. One major advantage of utilizing plant-based medicines is their generally safer profile than synthetic alternatives, offering significant therapeutic advantages at lower costs6. Plants also serve as sources of essential minerals such as calcium, iron, magnesium, and zinc, which are crucial for maintaining health. A lack of these micronutrients can lead to various deficiency disorders. Therefore, evaluating the nutrient content and proximate composition of medicinal plants is important for understanding their dietary significance and potential health benefits7. Nutrient analysis involves measuring key components such as ash, moisture, crude fat, crude fiber, and carbohydrates to evaluate their nutritional value8. These parameters help determine the plant’s potential as a food or nutraceutical source and support its traditional uses in local diets and medicine9,10.

Quantitative analysis is a scientific approach used to determine the concentration of specific compounds within a sample. In this study, advanced analytical techniques such as Ultra Performance Liquid Chromatography–Photodiode Array (UPLC-PDA) were employed for the accurate identification and quantification of phytochemicals. This method helps authenticate plant materials and assess their pharmacological relevance by detecting key bioactive compounds such as polyphenols, flavonoids, and alkaloids based on their UV absorbance and retention time11. Moreover, mineral analysis was conducted to determine the presence of essential elements like sodium (Na), potassium (K), calcium (Ca), magnesium (Mg), iron (Fe), zinc (Zn), copper (Cu), and manganese (Mn). These minerals play vital roles in plant metabolism and also contribute to the nutritional and therapeutic value of medicinal plants. Accurate mineral profiling helps evaluate their potential as nutraceuticals12. These analyses provide scientific validation of traditional uses and highlight the potential health benefits of medicinal plants. The main objective of the study was to examine the nutritional composition, mineral content, and phytochemical profile of selected medicinal plants commonly used by local communities.

Medicinal plants have long been integral to traditional healthcare systems, especially in rural and tribal communities where modern medical facilities are limited. Despite their widespread use, many of these plants remain scientifically underexplored in terms of their nutritional and phytochemical composition. This study is needed to bridge the gap between traditional knowledge and scientific validation by profiling essential minerals, nutrients, and bioactive compounds. Understanding these parameters not only supports the safe use of these plants but also aids in the development of standardized herbal formulations. In the future, these findings can contribute to the formulation of plant-based nutraceuticals, promote sustainable use of natural resources, and support public health initiatives addressing nutrient deficiencies. The present study aims to investigate the nutritional composition, phytochemical constituents, and quantitative profiling of selected medicinal plants from this ecologically significant region. Nutritional analysis and minerals provide insight into the dietary value and safety of these plants when consumed either as food or medicine. In addition to nutritional profiling, secondary phytochemical screening was conducted to identify the presence of bioactive compounds such as alkaloids, flavonoids, phenols, saponins, and tannins, which are responsible for therapeutic efficacy. Previous studies have highlighted the significance of medicinal flora from the Western Himalayas for their rich phytochemistry and biological activities; however, very few have focused on a combined approach of nutritional estimation, mineral quantification, and advanced phytochemical profiling. Therefore, this research not only contributes to the scientific validation of traditional medicinal practices but also aids in the conservation and sustainable utilization of plant biodiversity in the Renukaji region. The findings are expected to provide a baseline for future pharmacognostic and nutraceutical applications of these plants.

Materials and methods

Research area

The field-based ethnobotanical survey was conducted in Renukaji Wildlife Sanctuary (Sirmaur) in Himachal Pradesh from 2020 to 2023 (Fig. 1A, B). It is situated in the Sirmaur of Himachal Pradesh. Covering an area of 4.03 km², the sanctuary is positioned at an altitude ranging from 640 to 950 m above mean sea level. It spans the geographical coordinates of 30°35’58”− 30°37’08” N latitude and 77°26’34”–77°28’21” E longitude (Table 1). This sanctuary lies within the catchment area of the Giri River at Dadahu, near Nahan town, in the outer range of the Western Himalayas. The sanctuary’s landscape is characterized by the northern boundary covered with Sal (Shorea robusta) forests, gradually transitioning to sub-tropical pine (Pinus roxburghii) forests at higher altitudes. The map of the study site and geographical coordinates (altitude, latitude, and longitude) of ten villages surrounding the Wildlife Sanctuary were undertaken during the collection of plant samples from the study sites.

Fig. 1
figure 1

(A) Represents Renukaji Wildlife Sanctuary (*) in District Sirmaur. (B) Represents Sampling Sites in Renukaji Wildlife Sanctuary ( ).

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Table 1 Geographical coordinates of ten major villages of Renukaji wildlife sanctuary areas.
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Methodology

Plant collection and identification

The plant specimens were collected from various villages of Renukaji Wildlife Sanctuary of Sirmaur in Himachal Pradesh between 2020 and 2021 (Table 1). Information such as local names, growth habits, plant parts used, treated ailments, and methods of use was gathered through questionnaires and interactions with local healers, villagers, and farmers. The collected specimens were scientifically identified and dried for herbarium preparation. Each specimen was labeled with details including scientific name, local name, family, collection site, and medicinal uses. Identification was confirmed using reference specimens at the Botanical Survey of India (Directorate of Extension Education, UHF, Nauni, Solan, H.P.). Finally, the authenticated specimens were submitted to the herbarium at Shoolini University, Bajhol, Solan, where voucher numbers were assigned as shown in Table 2.

Nutrient analysis

Sample Preparation

The Leaf and root samples of ten species (Achyranthes aspera L., Aegle marmelos (L.) Correa, Azadirachta indica A. Juss., Berberis lycium Royle, Cassia fistula L., Senna tora (L.) Roxb, Senegalia catechu (L.f.) P.J.H.Hurter & Mobb., Syzygium cumini (L.) Skeels, Tinospora cordifolia (Willd.) Hook.f. & Thomson, Vitex negundo L.) were collected from Sanctuary areas located in the Sirmaur districts of Himachal Pradesh. For leaf collection, mature leaves were gathered, specifically selecting the fifth and sixth mature leaves from the top of each herb and shrub. These leaves were taken from four different directions. In the case of tree leaves, canopy leaves were hand-harvested from all four directions for each tree. For root collection, taproot was collected from sterilized pruning scissors dipped in 70% ethanol, which were used to excise out each root, approximately 9–12 cm in length. The gathered specimens were located inside paper bags and conveyed to the laboratory for additional examination. Subsequently, the samples were shade-dried for 15–20 days to ensure optimal results. After drying, the specimens were crushed in a grinder to obtain a fine powder for further biochemical analysis. The different parameters examined and the methods employed for their analysis are described in the following sections:

Ash

The determination of ash content was carried out using the procedure described by13. A porcelain crucible was subjected to drying at 106 °C for 1 h, subsequently cooled, and evaluated (W1). Following this, 3 g of finely crushed leaf sample were located within the crucible, after which it was re-weighed (W2). The crucible containing the plant specimens underwent ashing initially at 240 °C for an hour, followed by further ashing at 540 °C for six hours utilizing a muffle heating system. Following the ashing process, the leaf specimen was allowed to cool within a desiccator before being weighed once more (W3)13.

$$\:\:\:\text{A}\text{s}\text{h}\:\left(\text{\%}\right)\:=\frac{\text{W}2\:-\:\text{W}3\:}{\text{W}2\:-\:\text{W}1}\times\:100$$
Moisture

An immaculate container underwent oven drying at 105 °C for 1 h, subsequently cooled, and its weight was recorded as W1. A desiccated leaf specimen weighing 3 g (W2) was introduced into the flask and subjected to oven drying at 100 °C. Following this, the sample was cooled within a desiccation chamber and evaluated, yielding the measurement (W3)13. The moisture content of the fresh samples was determined using the procedure described by13.

$$\:\text{M}\text{o}\text{i}\text{s}\text{t}\text{u}\text{r}\text{e}\:\left(\text{\%}\right)\:=\frac{\text{W}2\:-\:\text{W}3\:}{\text{W}2\:-\:\text{W}1}\times\:100$$
Crude fat

The determination of crude fat was carried out using the procedure described by13. The 5 g of powdered sample underwent extraction in 100 mL of diethyl ether, shaken for 28 h using an orbital shaker. The resulting ether extract was gathered in a beaker previously weighed (W1). The filtrate obtained was directed into the same beaker post-equilibration with another 100 mL of diethyl ether, then shaken once more for 24 h, and again weighed (W1). The ether was subsequently evaporated to aridness in a steam bath and concentrated, followed by drying in an oven at 40–60 °C. Finally, the beaker was reweighed (W2)13.

$$\:\:\text{C}\text{r}\text{u}\text{d}\text{e}\:\text{f}\text{a}\text{t}\:\left(\text{\%}\right)\:=\frac{\text{W}2\:-\:\text{W}1}{\text{W}\text{e}\text{i}\text{g}\text{h}\text{t}\:\text{o}\text{f}\:\text{o}\text{r}\text{i}\text{g}\text{i}\text{n}\text{a}\text{l}\:\text{s}\text{a}\text{m}\text{p}\text{l}\text{e}\:}\times\:100$$
Crude fiber

A desiccated leaf sample weighing 2 g was treated with 100 mL of 1.25% H2SO4 for one hour, followed by filtration under pressure. The residue left after filtration was washed away with warm water. This procedure was then repeated on the residue using 100 mL of 1.25% NaOH solution. The remaining filtrate was dried at 95 °C and its weight (C1). Subsequently, it was incinerated in a muffle furnace at 540 °C for 4 h and weighed again, (C2)13.

$$\:\:\text{C}\text{r}\text{u}\text{d}\text{e}\:\text{f}\text{i}\text{b}\text{r}\text{e}\:\left(\text{\%}\right)\:=\frac{\text{C}2\:-\:\text{C}1}{\text{W}\text{e}\text{i}\text{g}\text{h}\text{t}\:\text{o}\text{f}\:\text{o}\text{r}\text{i}\text{g}\text{i}\text{n}\text{a}\text{l}\:\text{s}\text{a}\text{m}\text{p}\text{l}\text{e}\:\:\:}\times\:100$$
Crude protein

The determination was made using the Kjeldahl method13.

$$\:\text{N}\text{i}\text{t}\text{r}\text{o}\text{g}\text{e}\text{n}\:\text{c}\text{o}\text{n}\text{t}\text{e}\text{n}\text{t}\:\left(\text{\%}\right)\:=\frac{\left[\left(\text{m}\text{l}\:\text{s}\text{t}\text{a}\text{n}\text{d}\text{a}\text{r}\text{d}\:\text{a}\text{c}\text{i}\text{d}\times\:\text{N}\:\text{o}\text{f}\:\text{a}\text{c}\text{i}\text{d}\right)-\left(\text{m}\text{l}\:\text{b}\text{l}\text{a}\text{n}\text{k}\times\:\text{N}\:\text{o}\text{f}\:\text{b}\text{a}\text{s}\text{e}\right)\right]-(\text{m}\text{l}\:\text{s}\text{t}\text{d}\:\text{b}\text{a}\text{s}\text{e}\times\:\text{N}\:\text{o}\text{f}\:\text{b}\text{a}\text{s}\text{e})\times\:1.4007}{\text{W}\text{e}\text{i}\text{g}\text{h}\text{t}\:\text{o}\text{f}\:\text{s}\text{a}\text{m}\text{p}\text{l}\text{e}\:\text{i}\text{n}\:\text{g}\text{r}\text{a}\text{m}\text{s}}$$

In this context, with “N” representing normality, the percentage of crude protein was derived by multiplying the nitrogen value by a factor of 6.25.

\({\text{Crude protein }}\left( \% \right)\,=\,{\text{Nitrogen in sample}} \times {\text{6}}.{\text{25}}.\)

Carbohydrate

It was determined by subtracting the combined values of total ash, crude protein, crude fiber, and lipid from the total dry matter13,14.

\({\text{Carbohydrate }}\left( \% \right)\,=\,{\text{1}}00 - \left( {\% {\text{ Moisture }}+{\text{ }}\% {\text{ Ash }}+{\text{ }}\% {\text{ Crude Fat }}+{\text{ }}\% {\text{ Crude Fiber }}+{\text{ }}\% {\text{ Crude Protein}}} \right)\)

Mineral analysis

Sample preparation

The plant sample was cleansed using a method detailed by15, which included a sequence of rinses under tap water, treatment with 0.1 N HCl, and a final wash with distilled water. After this cleansing procedure, the plant samples were evenly laid out on filter paper sheets and left to air-dry in the shade for a duration of 15 to 20 days. Once thoroughly dried, the specimens were crushed, ground, and carefully preserved in polythene bags for later analysis of elements.

Analysis of mineral

Total Nitrogen (N) content in the leaf samples was determined by the micro-kjeldhal’s distillation method following the procedure described by the Association of Official Analytical Chemists (AOAC, 1975)16. Digestion of leaf samples in a 20 mL diacid mixture of HCl: HClO4 in the volumetric ratio of (4:1) and the final volume was 100 mL. Then the extract was utilized for the analysis of P, K, Ca, and S. Phosphorus in the digest was analyzed by the Canada-molybdate yellow color method17. Digestion of Potassium and calcium were analyzed by a flame photometer18. Sulfur was evaluated by the turbidimetric process19. Sodium, zinc, copper, iron, and manganese were determined by the atomic absorption spectrophotometric technique.

Phytochemical analysis

The Phytochemical analysis involves the identification of bioactive compounds present in plant materials. Typically, dried and powdered plant samples are subjected to solvent extraction using solvents like methanol, ethanol, or acetone. The extracts are then filtered and concentrated and then subjected to various standard chemical tests specific to each group of phytochemicals.

Alkaloid

5 g of dried leaf extract were measured and combined with 200 mL of a solution containing 10% acetic acid in ethanol. Afterward, the blend was covered and left undisturbed for 5 h. Subsequently, the blend underwent filtration, and the resulting filtrate was concentrated on a water bath until its volume reduced to one-fourth of the original. Concentrated NH4OH was introduced into the extract until precipitation was achieved. The solution was then subjected to washing with dilute NH4OH and filtered. The residue obtained was dried, weighed, and used to determine the alkaloid content13.

$$\:\text{A}\text{l}\text{k}\text{a}\text{l}\text{o}\text{i}\text{d}\:\left(\text{\%}\right)\:=\frac{\text{W}\text{e}\text{i}\text{g}\text{h}\text{t}\:\text{o}\text{f}\:\text{p}\text{r}\text{e}\text{c}\text{i}\text{p}\text{i}\text{t}\text{a}\text{t}\text{e}}{\text{W}\text{e}\text{i}\text{g}\text{h}\text{t}\:\text{o}\text{f}\:\text{o}\text{r}\text{i}\text{g}\text{i}\text{n}\text{a}\text{l}\:\text{s}\text{a}\text{m}\text{p}\text{l}\text{e}}\times\:100$$
Flavonoid

The process involved extracting 10 g of leaf plant powder repeatedly with 100 mL of 80% aqueous methanol over a span of 3 days. The entire solution was subsequently filtered through Whatman filter paper. The resulting filtrate was then transferred to a crucible and evaporated to dryness in a water bath until a constant weight was achieved. The weight obtained provided an estimation of the flavonoid content present in the plant leaf sample20.

$$\:\text{F}\text{l}\text{a}\text{v}\text{o}\text{n}\text{o}\text{i}\text{d}\:\left(\text{\%}\right)=\frac{Weight\:of\:dried\:extract\:}{Weight\:of\:sample}\times\:100$$
Saponin

5 g of dried leaf powder were combined with 50 mL of 20% ethanol, agitated on a shaker for 30 min, and then subjected to heating in a water bath at 55 °C for 4 h. The resultant mixture was filtered through Whatman filter paper. The residue collected was subjected to another extraction with 200 mL of 20% aqueous ethanol. The filtrates from both extractions were merged and concentrated to 40 mL in a water bath set at 90 °C. The concentrated solution was transferred into a separating funnel, to which 20 mL of diethyl ether ((C2H5)2O) was added and vigorously shaken. The upper layer of ether was removed, leaving the aqueous layer in a beaker. This aqueous layer was returned to the separating funnel, and 60 mL of n-butanol (C4H10O) were added and vigorously shaken. The upper layer of n-butanol was kept, and the lower layer was discarded. The n-butanol layer was rinsed twice with 10 mL of 5% aqueous sodium chloride. The resulting solution was gathered and evaporated in a water bath, then dried until a constant weight was reached at 40 °C in an oven13.

$$\:\text{S}\text{a}\text{p}\text{o}\text{n}\text{i}\text{n}\:\left(\text{\%}\right)\:=\frac{Weight\:of\:residue\:}{Weight\:of\:original\:sample}\times\:\:100$$

Analysis of polyphenols

Polyphenols of selected medicinal plant samples was conducted using the UPLC-PDA method employing the sonication procedure21. The presence of Polyphenol in the samples was confirmed by contrasting the UV spectra of standards with the retention duration of the samples. In the current study, three commonly used medicinal plants Achyranthes aspera L., Berberis lycium Royle and Vitex negundo L. gathered from the Renukaji Wildlife Sanctuary (Sirmaur) were analyzed for polyphenols and flavonoids using the UPLC-PDA method (Waters Acquity UPLC H-class).

Sample preparation

One gram of plant material was utilized for sample preparation. UPLC/LC-MS grade MeOH was employed for the extraction process. The sample was stirred for 6 min, followed by sonication for 25 min. Subsequently, it was centrifuged at 8000 rpm for 8 min, and the final volume collected was 10 mL. From this final volume, 1 mL was extracted and filtered using a 0.22 µ PTFE syringe filter. The filtered samples were then subjected to UPLC-PDA analysis.

Standard preparation

The selected phenolic acids/flavonoids, including P-coumaric acid, Gallic acid, Pro-catechuic acid, Vanillic acid, Rutin, Syringic acid, Caffeic acid, Epicatechin, EGCG, Ferulic acid, Quercetin, and Kaempferol standards, were utilized for analyses through UPLC-PDA (Waters Acquity UPLC, H-class). A standard stock solution was formulated with a concentration of 1 mg/mL. The standards were dissolved in MeOH (LC-MS grade), filtered through a 0.22 μm syringe filter, and kept at 4 °C for further analysis. Working solutions ranging from 2 ppm to 200 ppm were prepared through six serial dilutions of the calibration curve’s working solution. Identification of standards on UPLC-PDA was conducted based on their UV absorbance spectrum and retention times. The concentration of compounds in the sample was determined by comparing the sample peak areas to a standard calibration curve.

Quantification of phenolic acids

Quantification of selected phenolic acids and flavonoids was conducted using the Waters Acquity UPLC, H-class system16. The analytical column used was the ACE Ultra Core 2.5 Super C18 column (2.1 mm X 100 mm, 2.5 μm), with detection set at 270 nm. A gradient elution system was utilized, where mobile phase A consisted of 0.1% formic acid in water, and mobile phase B comprised 0.1% formic acid in ACN. The gradient started at 5% B and remained constant 0.3 min, then increased to 30% B from 0.3 min to 9 min, followed by a further increase to 70% B. Between 9 and 11 min, it was kept at 50% B and then maintained at 50% B until 12 min. At 12.2 min, the mobile phase returned to initial conditions, 5% B, and stayed until 16 min. The elution happened at a solvent flow rate of 0.30 mL/min. Quantification of compounds in the sample was accomplished by constructing a calibration curve and determining peak area by matching retention time and UV spectra.

Results

Ethnomedicinal uses

The indigenous inhabitants of Renukaji Wildlife Sanctuary, Sirmaur in Himachal Pradesh rely on locally sourced medicinal plants for their survival. These tribal communities have developed a lifestyle that is largely self-sustaining and culturally approved, centered around the utilization of various plant parts to treat various illnesses, with leaves being a commonly utilized component in medicine preparation. Conditions such as diabetes, jaundice, cold, cough, piles, skin disorders, and stomach problems are prevalent health issues that can be effectively addressed using these local herbal remedies. The medicinal plants were utilized, such as Achyranthes aspera L. leaves are utilized to cure cough and hemorrhoids, leaves extract of Aegle marmelos is used to control the cholesterol level, asthma, indigestion, and hepatitis, Azadirachta indica A. Juss. leaves are utilized to treat head lice, skin disorder, itching, diabetes, intestinal warm killing and blood purifier, root of Berberis lycium Royle is utilized to cure piles, diabetes and jaundice, Cassia fistula L. leaves is used to make decoction and taken for vomiting, nausea, abdominal pain and cramps, Senna tora (L.) Roxb. leaves are cooked and eaten for gastric problems, the leaves extract of Senegalia catechu (L.f.) P.J.H.Hurter & Mabb. is used for dysentery, mouth ulcer and piles, Syzygium cumini (L.) Skeels leaves and fruits are taken orally to treat diabetes and morning sickness, Stem juice of Tinospora cordifolia (Willd.) Hook.f. & Thomson is used to boost immunity, improve digestion, diabetes, jaundice, urinary problems and skin diseases, and extract of leaves Vitex negundo L. is employed on the skin to relieve muscle aches and joint pains (Table 2).

Table 2 Ethnomedicinal uses of plants.
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Mineral analysis

The evaluation of elements within the plants revealed their presence in all sampled plants. These elements play a crucial role in curing various types of diseases. In the present study, maximum nitrogen content was found in Azadirachta indica A. Juss. (3.12 ± 0.06) and minimum in Tinospora cordifolia (Willd.) Hook.f. & Thomson (0.44 ± 0.12), Potassium was found to be highest in Achyranthes aspera L. (1.46 ± 0.11) and lowest in Senegalia catechu (0.22 ± 0.11), Calcium was most abundant in Aegle marmelos (L.) Correa (3.68 ± 0.70) and least abundant in Cassia fistula L. (0.43 ± 0.23). Meanwhile, Phosphorus was most prevalent in Senna tora (L.) Roxb. (0.38 ± 0.15) and least prevalent in Syzygium cumini (L.) Skeels (0.15 ± 0.08), and Sulphur content was most abundant in Achyranthes aspera L. (0.37 ± 0.29) and least abundant in Senegalia catechu (L.f.) P.J.H.Hurter & Mabb. (0.11 ± 0.01) in Table 3; Fig. 2. The highest sodium content in ppm was found in Cassia fistula L. (398.00 ± 14.00), and the lowest was found in Senegalia catechu (L.f.) P.J.H.Hurter & Mabb. (256.33 ± 17.00). Iron was found to be highest in Tinospora cordifolia (Willd.) Hook.f. & Thomson (537.66 ± 10.26) and lowest in Aegle marmelos (L.) Correa (164.00 ± 17.05). Manganese was most abundant in Cassia fistula L. (94.43 ± 0.87) and least abundant in Senegalia catechu (L.f.) P.J.H.Hurter & Mabb. (6.86 ± 0.95). Copper was found to be highest in Cassia fistula L. (28.92 ± 0.89) and lowest in Senegalia catechu (L.f.) P.J.H.Hurter & Mabb. (6.50 ± 0.88). Zinc content was most abundant in Berberis lycium Royle (86.98 ± 0.90) and least abundant in Senegalia catechu (L.f.) P.J.H.Hurter & Mabb. (12.39 ± 0.42) in Table 4.

Table 3 Mineral analysis (%) of ethnomedicinal plants (Mean ± SD).
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Fig. 2
figure 2

Mineral analysis of medicinal plants (Mean ± SD).

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Table 4 Mineral analysis (ppm) of ethnomedicinal plants (Mean ± SD).
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Nutrient analysis

The moisture content varied, with Vitex negundo L. displaying the highest level at 14.803 ± 1.153 and Senna tora (L.) Roxb. exhibiting the lowest at 4.303 ± 0.867. Ash content peaked in Senegalia catechu (L.f.) P.J.H.Hurter & Mabb. at 15.632 ± 0.872 and was lowest in Syzygium cumini (L.) Skeels at 3.384 ± 0.785. Vitex negundo L. had the highest crude fat content at 6.601 ± 0.676, while Aegle marmelos (L.) Correa showed the lowest at 0.827 ± 0.344. Senna tora (L.) Roxb. recorded the highest crude fiber content at 17.343 ± 0.763, whereas Vitex negundo L. displayed the lowest at 5.399 ± 0.524. Cassia fistula L. presented the highest crude protein content at 26.328 ± 0.614, whereas Syzygium cumini (L.) Skeels had the lowest at 1.528 ± 0.136, For carbohydrate content, Syzygium cumini (L.) Skeels exhibited the highest value at 79.237 ± 1.233, whereas Senegalia catechu (L.f.) P.J.H.Hurter & Mabb. displayed the lowest at 28.946 ± 0.243 in Table 5; Fig. 3.

Table 5 Nutrient content (%) of ethnomedicinal plants (Mean ± SD).
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Fig. 3
figure 3

Nutrient analysis of ethnomedicinal plants (Mean ± SD).

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Phytochemical analysis

The phytochemical composition of selected medicinal plants, gathered from the Renukaji Wildlife Sanctuary (Sirmaur), was examined. In the present investigation, the alkaloid content was highest in Aegle marmelos (L.) Correa at 0.493 ± 0.035 and lowest in Azadirachta indica A. Juss. at 0.353 ± 0.100. Flavonoid levels peaked in Cassia fistula L. at 0.630 ± 0.100 and were lowest in Berberis lycium Royle at 0.343 ± 0.110. Saponin content was highest in Aegle marmelos (L.) Correa at 1.530 ± 0.100 and lowest in Syzygium cumini (L.) Skeels at 1.253 ± 0.115 in Table 6; Fig. 4.

Table 6 Phytochemical analysis (%) of ethnomedicinal plants (Mean ± SD).
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Fig. 4
figure 4

Phytochemical analysis of medicinal herbs (Mean ± SD).

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Quantitative analysis of selected ethnomedicinal plants from Renukaji wildlife sanctuary

In the current study, the quantitative analysis of medicinal plants from Renukaji Wildlife Sanctuary (Sirmaur) was carried out using the UPLC - PDA method. From the sanctuary area, commonly used medicinal plants, viz. Berberis lycium Royle, Ahyranthes aspera L., and Vitex negundo L. were collected for quantitative analysis due to their wide availability and frequent use by local communities for medicinal purposes. Their accessibility made it feasible to collect sufficient plant material for detailed phytochemical and mineral analysis. The purpose of studying their nutraceutical value is to confirm their traditional uses, understand their nutritional and medicinal benefits, and encourage their use as natural health supplements while also supporting their conservation and further examining for the presence of polyphenols and flavonoids in methanolic extract using 270 nm wavelength. The consequences of the UPLC analysis are given in Fig. 5.

Fig. 5
figure 5

Chromatogram of standard mixture of polyphenol carried out using UPLC analysis. (1) Gallic acid, (2) Pro-catechuic acid, (3) Vanillic acid, (4) Syringic acid, (5) Caffeic acid, (6) EC (Epicatechin), (7) EGCG, (8) P-coumaric acid, (9) Ferulic acid, (10) Rutin, 11. Quercetin, 12. Kaempferol.

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Berberis lycium royle

The UPLC analysis indicated the presence of Gallic acid and EC (Epicatechin) in the methanolic extract of Berberis lycium Royle collected from three distinct Wildlife Sanctuaries in Himachal Pradesh in Fig. 6A. The data revealed that the roots of Berberis lycium Royle collected from Renukaji Wildlife Sanctuary exhibited the EC (Epicatechin) content (4.187 µg/10 mg) and Gallic acid (0.147 µg/10 mg). The presence of Gallic acid and Epicatechin (EC) in the methanolic extract of Berberis lycium Royle roots, as identified through UPLC analysis, is supported by existing literature on the phytochemistry of this species. Both compounds are well-known phenolic metabolites commonly found in medicinal plants. Gallic acid is a phenolic acid with strong antioxidant and antimicrobial properties, while Epicatechin is a flavonoid known for its antioxidant, anti-inflammatory, and cardioprotective effects. Their occurrence in Berberis lycium aligns with the plant’s traditional medicinal use. The variation in their concentrations, particularly the higher EC content in samples from Renukaji Wildlife Sanctuary, may be attributed to environmental and ecological differences influencing secondary metabolite synthesis.

Achyranthes aspera L.

The UPLC analysis revealed the presence of Gallic acid, Pro-catechuic acid, Ferulic acid, P-coumaric acid, and Rutin in the methanolic extract of Achyranthes aspera L. collected from Renukaji Wildlife Sanctuaries (Sirmaur) in Himachal Pradesh in Fig. 6B. The data indicated that in the leaves of Achyranthes aspera L. collected from Renukaji Wildlife Sanctuary, the Rutin content (1.267 µg/10 mg), Gallic acid (0.183 µg/10 mg), Pro-catechuic acid (0.182 µg/10 mg), P-coumaric acid (0.623 µg/10 mg), Ferulic acid (0.080 µg/10 mg) was observed. The UPLC analysis of Achyranthes aspera L. leaf extract collected from Renukaji Wildlife Sanctuary in Himachal Pradesh confirmed the presence of several important secondary metabolites, including Rutin, Gallic acid, Protocatechuic acid, P-coumaric acid, and Ferulic acid. These compounds are well-known phenolic and flavonoid constituents that play vital roles in the plant’s defense mechanisms and therapeutic potential. Rutin, a potent flavonoid (1.267 µg/10 mg), is known for its strong antioxidant, anti-inflammatory, and vascular-protective effects. Gallic acid (0.183 µg/10 mg) and Protocatechuic acid (0.182 µg/10 mg) are phenolic acids recognized for their antimicrobial and radical-scavenging properties. P-coumaric acid (0.623 µg/10 mg) and Ferulic acid (0.080 µg/10 mg) are hydroxycinnamic acids that contribute to the plant’s antioxidant capacity and may help modulate various metabolic pathways. The presence of these bioactive compounds supports the traditional medicinal uses of Achyranthes aspera and highlights its pharmacological significance.

Vitex negundo L.

The UPLC analysis indicated the presence of Pro-catechuic acid and Vanillic acid in the methanolic extract of Vitex negundo L. collected from Wildlife Sanctuaries in Fig. 6C. The data revealed that in the leaves of Vitex negundo L., the Pro-catechuic acid content (4.156 µg/10 mg) was present and Vanillic acid is not quantified. The UPLC analysis confirmed the presence of Protocatechuic acid and Vanillic acid in the methanolic leaf extract of Vitex negundo L. collected from Wildlife Sanctuaries, as shown in Fig. 6C. Protocatechuic acid was detected at a concentration of 4.156 µg/10 mg, whereas Vanillic acid was detected but not quantified. Both compounds are known phenolic acids with significant pharmacological properties. Protocatechuic acid is widely recognized for its antioxidant, anti-inflammatory, and antimicrobial activities, contributing to the plant’s traditional therapeutic uses. The detection of Vanillic acid, even in trace amounts, supports the presence of diverse bioactive phenolics in Vitex negundo, reinforcing its medicinal value in traditional and modern herbal formulations.

Fig. 6
figure 6

Chromatogram of commonly used plants: (a) Roots of Berberis lycium Royle, (b) Leaves of Achyranthes aspera L., (c) Leaves of Vitex negundo L. analyzed using UPLC analysis collected from Renukaji Wildlife Sanctuary.

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Discussion

In this study, analyzing medicinal plants for their mineral, nutrient, and phytochemical content is crucial for understanding their therapeutic potential and ensuring their safe and effective use in traditional and modern medicine. This comprehensive approach bridges the gap between traditional knowledge and scientific validation, allowing for a better understanding of the pharmacological basis of plant-based treatments. Minerals are crucial nutrients that play vital roles in numerous physiological processes in the human body. Analyzing the mineral content of medicinal plants helps in determining their nutritional value and potential health benefits. Moreover, mineral profiling contributes to identifying plants that can help address micronutrient deficiencies, particularly in undernourished populations. Common minerals analyzed in medicinal plants include calcium, phosphorus, potassium, sulfur, and nitrogen. These minerals contribute to bone health, immune function, enzyme activation, and antioxidant defense mechanisms22. Nutrient analysis involves determining the presence and concentration of moisture, ash, carbohydrates, proteins, fiber, and fats in medicinal plants. Such data is essential in assessing the overall nutritional profile and dietary relevance of these plant species. Carbohydrates provide energy, proteins are crucial for tissue restoration and development, and Fats act as an energy source and assist in absorbing nutrients23. Fiber aids digestion and contributes to gastrointestinal health, while moisture and ash content reflect the plant’s freshness and total mineral content, respectively. Phytochemical analysis proves highly valuable for assessing the active biological compounds present in medicinal plants. Phytoconstituents are bioactive compounds found in vegetation that have medicinal properties and contribute to their therapeutic effects24. These include flavonoids, alkaloids, and saponins. Phytochemicals exhibit antioxidant, antimicrobial, and anticancer activities25. Their diverse biological activities support the use of medicinal plants in treating a wide range of health conditions. Analyzing the phytochemical profile of medicinal plants helps in identifying their active constituents and understanding their mechanisms of action26. The diverse range of phytochemicals and elemental makeup present in plants significantly influences their effectiveness for medicinal purposes. It also assists in the standardization and quality control of herbal formulations, which is necessary for their acceptance in clinical practices. Therefore, it is crucial to examine the phytochemicals, and elements present in medicinal herbs to assess their healing capabilities. Herbs contain all the essential components necessary for human health, making them valuable sources of nutrition and therapeutic remedies27. This dual role enhances their significance in the development of functional foods and nutraceuticals. While the utilization of medicinal plants dates back to ancient times, their popularity has surged in modern years. In the present investigation, the ash, crude protein, and carbohydrate levels were assessed in Berberis lycium L. The ash content in Berberis lycium L. was measured at 5.481%, crude protein at 4.769, and carbohydrate at 59.886%. However, a study conducted in the Western Himalayas on Berberis lycium L. reported ash, crude protein, and carbohydrate levels at 5.40%, 5.84%%, and 27.23%, respectively28. Notably, the ash content was lowest in Berberis lycium L. in the Western Himalaya study, contrasting with our findings of 5.481%. Our study observed a lower crude protein percentage compared to the Western Himalaya study, while the carbohydrate percentage was higher in our study. Such variation could be attributed to differences in environmental conditions, altitude, soil composition, and seasonal factors, which are known to influence the biochemical composition of plants.

Quantitative characterization involves determining the concentration of specific compounds or groups of compounds in medicinal plants. This could include quantifying individual minerals, nutrients, or phytochemicals present in the plant material29. This type of analysis plays a vital role in ensuring consistency in herbal medicine, as the efficacy of these treatments depends on the presence of bioactive compounds in adequate quantities. Quantitative analysis provides valuable information about the potency and consistency of medicinal plants, enabling the standardization of herbal preparations for therapeutic use30. The research revealed that the leaves of the selected plants contain abundant phytochemicals and minerals, indicating their potential as valuable sources for pharmaceutical purposes. Such findings validate the ethnobotanical uses of these plants and open avenues for the discovery of new drug candidates derived from natural sources. Given the substantial presence of easily accessible phyto-constituents and minerals in these plants, they could serve as an effective means to supplement nutrients that are deficient or lacking elsewhere. This supports their integration into dietary interventions and alternative healthcare strategies, especially in resource-limited settings.

Conclusion

The study results indicate that medicinal plants in the research area contain a significant number of phytochemicals and minerals. Mineral, nutrient, and phytochemical analysis, along with quantitative characterization, are integral to evaluating the medicinal properties and safety of herbs utilized in folk and complementary drugs. The presence of essential minerals such as calcium, magnesium, potassium, and iron underscores their importance in promoting overall health and vitality. Moreover, the diverse range of nutrients contributes to their nutritional value and supports various physiological functions within the human body. Furthermore, the abundance of phytochemicals highlights their role in exerting antioxidant, and anticancer properties, thus offering promising avenues for the development of novel pharmaceuticals and nutraceuticals. Harnessing the curative potential of these medicinal herbs not only offers a natural alternative to conventional medicine but also underscores the importance of preserving biodiversity for the advancement of healthcare and wellness. Continued research into the synergistic interactions between these bioactive compounds will undoubtedly pave the way for the discovery of new therapeutic agents and the promotion of holistic approaches to health and well-being.