Research Article

Unlocking the Phytochemical Wealth of Berberis aristata: An Endangered Himalayan Medicinal Shrub

Nisha Tariq, A.R. Malik*, P.A. Sofi, Ishtiyak Ahmad Peerzada, Amerjeet Singh, Tahir Mushtaq, Nazir A. Pala, Mehvish Mushtaq, Moamenla Jamir and Peerzada Tabish Fayaz

Faculty of Forestry, Sher-e-Kashmir University of Agricultural Science and Technology Shalimar Srinagar Kashmir Campus, Benhama Ganderbal -191 201, India.

Received | May 14, 2026; Accepted | June 23, 2026; Published | July 22, 2026

*Correspondence | A.R. Malik, Faculty of Forestry, Sher-e-Kashmir University of Agricultural Science and Technology Shalimar Srinagar Kashmir Campus, Benhama Ganderbal -191 201, India; Email: [email protected]

Citation | Tariq, N., A.R. Malik, P.A. Sofi, I.A. Peerzada, A. Singh, T. Mushtaq, N.A. Pala, M. Mushtaq, M. Jamir and P.T. Fayaz. 2026. Unlocking the phytochemical wealth of Berberis aristata: An endangered Himalayan medicinal shrub. Journal of Plant Health, Ecology and Evolution, 2(1): 24-34.

DOI | https://dx.doi.org/10.17582/journal.pjar/2026/2.1.24.34

Keywords | Berberis aristata, Phytochemical profiling, Extraction yield, Secondary metabolites, Spectrophotometric analysis, Medicinal plants, Sustainable utilization

Copyright: 2026 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

Introduction

The Himalayan region is recognized as one of the hotspots of global biodiversity, where ecological, phytogeographical and evolutionary factors favor species richness. The bio-geographically unique nature of the region supports the maximum number of endemic species, harbouring about 18, 440 species of plants, of which 25.3% are endemic to the Asian region, a high proportion of which possesses unique medicinal properties (Malik et al., 2024a). India is a storehouse of a wide spectrum of ethno-medicinally important plant species and is one of the 18 mega biodiversity with more than 2500 plant species applied in various alternative and complementary medicines of medicinal importance (Mipun et al., 2023a). Berberis belongs to the dicotyledonous family; Berberidaceae and is the largest genus among the 16 genera, constituting about 70-80% of total species in the family (Khan et al., 2016a). Berberis has approximately 500 species distributed across the globe (Europe, Siberia, China, Afghanistan, India, Nepal and North-South America) out of which 60 species have been reported from the Indian Himalayas (Kaur et al., 2023). In homeopathy and ethnic medicine, the species have been used as medicine for their anti-inflammatory, anti-bacterial, anti-cancer, antiarrhythmic, hypoglycemic and hepatoprotective properties (Khan et al., 2016a). Berberis aristata commonly known as “Daru Haldhi, Kashmoi, Kashmal, Chitra, Kawdachh” is an endangered spinous medicinal shrub of western Himalayan region occurring in rare and sporadic conditions at an altitudinal range between 1800-2800 m above mean sea level (Thakur et al., 2019; Nisha, 2023a). The plant is widely distributed from the western Himalayas to Srilanka, Bhutan and hilly areas of Nepal (Choudhary et al., 2021). It is also found in the Nilgiri hills in south India (Sharma et al., 2018). The plant has glossy dark green and ovate leaves, stalked flowers and woody, yellowish brown roots with a thin covering of bark (Yadav, 2018a). Fruits are globose to ovoid; usually cover with bloom as in plums. The fruit colour is aconite violet (Komal et al., 2011a). Berberis aristata has been recognized for its pharmaceutical and medicinal properties (Alvarez et al., 2009; Nisha, 2023). It is also used in the printing and dyeing industry (Malik et al., 2024a). The roots, stems, leaves and fruits of Berberis aristata are traditionally used to treat wounds, diabetes, inflammations and jaundice. The extracts have been reported for antibacterial, antiviral, antifungal, antiarrhythmic, anticancer, anti-inflammatory and antidiabetic profiles (Yadav, 2018). The major chemical constituent of Berberis aristata is berberine, a quaternary isoquinoline alkaloid that is typically found in the roots and stems with numerous pharmacological activities (Komal et al., 2011a). A very valuable ayurvedic preparation ‘Rasaut’ an extract of either leaves, stem or root is prepared from this plant (Moin et al., 2023a). It is utilized as a blood cleaner, tonic and purgative for kids. It is also prescribed for skin conditions and jaundice (Alamzeb et al., 2013). The unbated exploitation of B. aristata roots and stem for its berberine alkaloid (Verma et al., 2017) and the non-availability of propagation protocol, agronomic ignorance, habitat loss, lack of motivation among stakeholders and complete dependence on natural population have rendered the species under immense threat in nature and IUCN has categorized it as endangered species of Indian Himalayan Region (Ali et al., 2008).

Although Berberis aristata is widely valued for its root-derived phytochemicals, its predominant use has led to extensive root harvesting, raising concerns regarding its conservation status. Despite this, limited attention has been given to the comparative evaluation of phytochemical constituents in other plant parts, particularly leaves and stem bark, which may serve as potential alternatives. This gap restricts the development of sustainable utilization strategies and highlights the need for comprehensive quantitative assessment of phytochemical constituents across different plant parts. Therefore, the present study reports a comparative and standardized quantitative assessment of major phytochemical constituents in the root, leaves and stem bark of Berberis aristata.

Materials and Methods

The root, stem bark, and leaves of Berberis aristata DC. were collected from Sindh Forest Range, Ganderbal, Jammu and Kashmir, during the autumn season (October–December). The plant material was collected in duplicate to minimize sampling errors. The collected specimens were identified and authenticated at the Centre for Biodiversity and Taxonomy, Department of Botany, University of Kashmir, Hazratbal, Kashmir. Proper voucher specimens were prepared and deposited in the herbarium of the Department of Botany, University of Kashmir, under Voucher No. 9354-KASH. Fresh root, stem bark, and leaf samples were shade dried at room temperature (20 ± 2°C) and pulverized into coarse powder. The powdered material of each plant part was successively extracted with ethanol, methanol, and distilled water using a mechanical shaker. The extracts obtained were filtered and concentrated under reduced pressure using a rotary evaporator with a water bath maintained at 50°C. The concentrated extracts were transferred into sterile vials and stored in a refrigerator at 4°C until further use.

Physicochemical analysis

Physicochemical analysis of root, stem bark, and leaf samples was carried out following standard procedures described in the Ayurvedic Pharmacopoeia of India (Government of India, 2007) and WHO guidelines for medicinal plant materials (WHO, 1998).

Total ash content was calculated using the formula:

Moisture content was determined as:

Acid insoluble ash was determined as:

Qualitative analysis

The plant extracts obtained by using different solvent extraction process and is subjected to different phytochemical tests to identify the plant constituents by using standard following methods Sofowara (1993) and Harborne and William (1973).

Quantitative estimation

Total flavonoid content (TFC): Using a colorimetric technique with the aluminium chloride, total flavonoid contents were determined (Kumar et al., 2008). Aqueous and ethanolic extracts that were adjusted to fall within the linearity range i.e. (400μg/ml), or Aliquots of extract solutions were taken and made up the volume 3ml with methanol and various dilutions of standard solution of the standard solution of Quercetin (10-100μg/ml) were added to 10ml volumetric flask. The aforementioned combination received 0.3ml of 5% NaNO2. 0.3ml of 10% AlCl3 was added after 5 minutes. After 6 min, 2ml of 1 M NaOH was added and distilled water was used to get the volume up to 10ml. The absorbance was then measured at 520 nm in comparison to a freshly made reagent blank

Test	Procedure	Observations (Indicating positive test)
Detection of Carbohydrates
Benedict’s test	0.5 mL filtrate + 0.5 mL Benedict’s reagent; heated in boiling water bath for 2 min	red precipitate
Detection of protiens
Millon’s and Ninhydrin test	1 mL plant sample + 1 mL 40% NaOH + few drops CuSO₄	violet/pink color
Detection of Alkaloids
Dragendroff’s	Few mL filtrate + 1-2 mL Dragendroff’s reagents	A reddish-brown precipitate
Detection of Tannins
Ferric chloride test	1mL filtrate + 3mL distilled water + 3 drops 10% Ferric chloride solution	Blue green colour
Detection of saponins
Foam test	0.5gm plant extract + 2mL water (vigorously shaken)	Persistent foam for 10 min
Detection of Flavonoids
Ammonia test	Few mL aqueous filtrate + conc. H₂SO₄ + 5 mL dilute NH₃	yellow color
Detection of phenolic compounds
Ferric cyanide test	3 mL ethanolic extract + few drops ferric cyanide (warm in water bath)	greenish to black color
Detection of terpenoids
Salkowski test	5 mL extract + 2 mL chloroform + 3 mL conc. H₂SO₄	reddish-brown
Detection of phytosterols
Acetic anhydride test	0.5 g ethanolic extract + 2 mL conc. H₂SO₄ + 2 mL acetic anhydride	violet/blue/green colour

 

after the solution had been well mixed. The extracts total flavonoid concentration was calculated as a percentage of Quercetin equivalent per 100 g dry weight of sample.

Total phenols content (TPC)

Total phenolic content was determined using the Folin–Ciocalteu colorimetric method as described by Singleton and Rossi (1965). An aliquot of the plant extract was mixed with diluted Folin–Ciocalteu reagent and allowed to react for a few minutes. Subsequently, sodium carbonate solution was added to the mixture to neutralize the reaction and facilitate color development. The reaction mixture was then incubated at room temperature for a specified period to allow the formation of a blue-colored complex. The intensity of the color, which is proportional to the phenolic content present in the sample, was measured spectrophotometrically at 765 nm against a suitable blank. Gallic acid was used as the standard for calibration, and the results were expressed as milligrams of gallic acid equivalents (mg GAE/g) of the sample.

Total alkaloid content (TAC)

The methodology of Harborne (1973) was employed to determine alkaloid in the species. 200 ml of 10% acetic acid in ethanol was added to 5 g of the sample, which was then weighed into a 250 ml beaker. The mixture was then covered and left to stand for 4 h. After filtering, the extract was concentrated on a water bath to one-quarter of the original volume. Concentrated ammonium hydroxide was added drop by drop to the extract. After allowing the whole solution to settle, the precipitated material was collected, cleaned with dilute ammonium hydroxide and then filtered. The alkaloid, which was dried and weighed, is the residual.

Total tannin content (TTC)

A plastic bottle measuring 50 ml was filled with 500 mg of the sample. A mechanical shaker was used to mix in 50 ml of distilled water for an hour. This was built up to specification and filtered into a 50 ml volumetric flask. Then, 2 ml of 0.1 M FeCl3 in 0.1N HCl and 0.008 M potassium ferro cyanide were added to 5 ml of the filtered and pipetted into a test tube. Within 10 minutes, the absorbance at 700 nm was determined. The determination of tannin was done by Van-Buren and Robinson (1981) methodology.

Total saponin content (TSC)

The Obadani and Ochuko (2002) methodology was adopted. 100 cm3 of 20% aqueous ethanol and 20 g of each sample were added to a conical flask after the samples had been grounded. The samples were heated over a hot water bath for 4 h while being continuously stirred at about 550˚C. Filtering the mixture allowed the residue to be extracted again using 200 ml of 20% ethanol. Over a water bath at around 90 ˚C, the mixed extracts were reduced to 40 ml, transferring the concentration into a 250 ml separatory funnel, 20 ml of diethyl ether was added and shaken vigorously. While the ether layer was discarded, the aqueous layer was recovered. The purification process was repeated. n-butanol 60 ml was added. Two separate washes of 10 ml of 5% aqueous sodium chloride were performed on the combined n-butanol extracts. In a water bath, the residual solution was warmed. The samples were dried in oven to a constant temperature following evaporation.

Statistical analysis

All data are expressed as mean ± SD (n= 3). Differences among plant parts were assessed by one-way ANOVA followed by Tukey’s HSD post-hoc test (α = 0.05).

Results and Discussion

Extraction yield

The extraction yield is a measure of the solvent efficiency to extract specific components from the original material. The results were reported in Table 1 and illustrated in Figure 1. Our results showed that maximum percent yield was obtained in leaves (5.0%), when Berberis aristata was extracted by methanol with followed by root (4.6%) and stem bark (4.6%). While the Ethanol extraction showed comparatively lower yields ranging with leaves (4.0%), followed by root (2.6%) and stem bark (2.0%). B. aristata when extracted by aqueous solvent gave the minimum yield per centage with leaves and root (3.8%) followed by stem bark (2.6%). Our results showed that methanol was efficient in extracting phytochemicals more than other solvents attributed to its higher polarity and better solubilizing capacity for a wider range of phytochemicals in the species. The same results have been seen in other Berberis species, as well (Bhatt et al., 2012). These findings are in agreement with previous studies that have shown that solvents like methanol, which are polar solvents, are more efficient at extracting the bioactive compounds from the other plant materials (Ncube et al., 2008).

 

Table 1: Percentage extraction yield of different solvent extracts from various plant parts of B.aristata.

Solvent	Plant part	Extraction yield (%)
Ethanol	Stem bark	2.0
	Leaves	4.0
	Root	2.6
Methanol	Stem bark	4.6
	Leaves	5.0
	Root	4.6
Aqueous	Stem bark	2.6
	Leaves	3.8
	Root	3.8

 

Physicochemical parameters

The results of physicochemical properties of B. aristata shown in Table 2 reveals that the ash content of the various plant sections was comparatively similar among different parts of the species, with values of 2.17%, 2.19%, and 2.11%, respectively for the stem bark, leaves, and root. However, the leaves had the highest content of acid insoluble ash (0.62%), followed by roots (0.50%) and stem bark (0.39%). Similarly, the leaves had the highest moisture content (76.00%), followed by stem bark (63.04%) and root (52.38%). The observed results of moisture content recorded for B. aristata leaves in this study is can be compared to that reported by Chauhan et al. (2014), who found values between 58–78% in fresh leaves of different Berberis species.

Preliminary phytochemical screening

The preliminary qualitative screening of the extracts is presented in Table 3 and showed that alkaloids were present with the well-documented presence of berberine, an isoquinoline alkaloid, as the principal bioactive constituent of B. aristata (Katiyar et al., 2012). Tannins and saponins were the most common compounds identified in most of the extracts especially the methanol and aqueous fractions which is expected since they are polar compounds. Flavonoids were found to be present in extracts of stem bark and leaf. Flavonoids have been reported to possess antioxidant and anti-inflammatory properties in many studies (Kumar and Pandey, 2013). Some metabolites might not have been found in the aqueous extract because of polarity; water is not able to dissolve polar compounds well. These findings justify the reason for pharmaceutical application of B. aristata and hence validating its use. Similar results were observed in previous phytochemical screenings conducted on the same plant species (Sutalangka et al., 2013).

 

Table 2: Comparison of physiochemical parameters of different plant parts of Berberis aristata.

Parameter	Stem bark (% w/w)	Leaves (% w/w)	Root (% w/w)
Total ash content	2.17	2.19	2.11
Acid insoluble ash	0.39	0.62	0.50
Moisture content	63.04	76.00	52.38

 

Quantitative analysis

Total alkaloid content: The results of the quantitative evaluation of alkaloids, phenols, tannins, flavonoids and saponins are presented in Table 4. Alkaloids are important group of secondary metabolites of Berberis genus, of which previous studies attributed some of the medicinal properties of barberry to the presence of these compounds (Zhaleh et al., 2025). In fact, barberry is known to be a rich source of various types of alkaloids with antioxidant and therapeutic

 

Table 3: Preliminary phytochemical screening of different solvent extracts of Berberis aristata.

Metabolite	SB-E	SB-M	SB-A	L-E	L-M	L-A	R-E	R-M	R-A
Carbohydrates	+	+	-	-	-	+	+	+	-
Proteins	-	-	+	-	-	+	+	-	+
Alkaloids	+	+	-	+	-	-	+	+	+
Tannins	-	+	+	+	+	+	+	+	+
Saponins	-	+	+	+	+	+	+	+	+
Flavonoids	+	+	-	+	-	-	-	+	+
Phytosterols	+	+	+	-	-	-	+	+	-
Terpenoids	+	+	+	-	-	+	-	+	-
Polyphenols	-	+	-	+	-	-	-	+	-

 

SB= Stem bark, L= Leaves, R= Root, E= Ethanol, M= Methanol, A= Aqueous (+) Presence, (-) Absence

 

Table 4: Quantitative phytochemical content (mean ± SD, n = 3) of leaves, stem bark, and roots of Berberis aristata.

Compound	Leaf	Bark	Root	Anova
Alkaloid (mg AE/g)	10.80 ± 1.20a	14.36 ± 0.54b	18.75 ± 0.65c	F = 99.38 p < 0.001
Tannin (mg GAE/g)	185.25 ± 1.75c	34.79 ± 0.30a	109.79±0.29b	F= 23606.32 p < 0.001
Total saponin content (mg DE/g)	8.96±0.24a	38.78±0.82b	11.56±2.2a	F = 661.23 p < 0.001
Total flavonoid content (mg QE/g)	54.95±1.2c	9.69±0.3a	35.27±0.6b	F = 3678.70 p < 0.001
Total phenol content (mg GAE/g)	62.23±0.63c	4.23±0.9a	29.15±1.4b	F = 3608.71 p < 0.001

 

Different lowercase letters within a row indicate significant differences among plant parts (Tukey’s HSD, α = 0.05). AE = atropine equivalent; GAE = gallic acid equivalent; DE = diosgenin equivalent; QE = quercetin equivalent.

 

 

 

properties (Och et al., 2021). Berberine is considered one of the main alkaloids of Berberis genus, many studies have focused on this valuable alkaloid in various Berberis species. Garhwal et al. (2010), by studying B. asiatica, B. aristata and B. lycium, revealed that the berberine content of these three species is different and also that their roots contain much more berberine content compared to the stem. The results of the current study display that total alkaloid content differed significantly among the three plant parts (F= 99.38, p < 0.001; Table 4). The highest content of total alkaloid was observed in the root (18.75 ± 0.65 mg AE/g), followed by bark (14.36 ± 0.54 mg AE/g), while leaves showed the lowest content (10.8 ± 1.2 mg AE/g) of total alkaloid (Figure 2). All three plant parts were significantly different from one another (Tukey’s HSD, p < 0.01; CLD: c, b, a for roots, bark, and leaves, respectively). This distribution corroborates with previous studies reporting maximum alkaloid accumulation in roots and stem bark of Berberis species due to the localization of berberine and related isoquinoline alkaloids in ground tissues (Imenshahidi and Hosseinzadeh, 2016; Zhaleh et al., 2025).

Total tannin content

The term “tannin” originates from the application of these compounds to tan animal hide into leather (Pizzi, 2021). Tannins are naturally occurring substances which can be found in almost all parts of most plant life. These compounds have been shown to play an important part in protecting the plant against fungal, bacterial and insect attacks, and also aid in maintaining plant health when water availability decreases during times of drought (Pizzi and Cameron, 1986). In the present study, tannin content showed the greatest absolute variation among the phytochemicals examined (F₂,₆ = 23606.32, p < 0.001; Leaves (185.25 ± 1.75 mg GAE/g) exhibited the highest level of tannins within B. aristata, followed by the tannin content of roots (109.79 ± 0.29 mg GAE/g), and the lowest tannin content was detected in the bark (34.79 ± 0.30 mg GAE/g) (Figure 3). All three plant parts differed significantly from one another (Tukey’s HSD, p < 0.001; CLD: c, a, b for leaves, bark, and roots, respectively). Although the levels of tannins appear to be relatively low in most species of Berberis, the B. aristata root samples (10.97%) were found to contain significantly higher percentages of tannins compared to that found in B. microphylla (9.53%), B. asiatica (1.7%), B. chitria (0.73%), and B. lycium (0.96%) (Srivastava et al., 2013b). Tannins within the Berberis genus are primarily used for their pharmacological and therapeutic benefits, such as being both astringents and antimicrobials, in traditional medicinal practices including Ayurveda (Sharma et al., 2024a). Therefore, given the significant amount of tannins present throughout the various parts of B. aristata, it could represent a strong natural compound with significant potential for the development of new pharmaceutical products.

 

 

Total saponin content

A significant effect of plant part on saponin content was observed (F = 661.23, p < 0.001; Table 4) Stem bark contained the highest total saponin content (38.78 ± 0.82 mg DE/g), substantially exceeding root (11.56 ± 2.2 mg DE/g) and leaf (8.96 ± 0.24 mg DE/g) tissues (Figure 4). Stem bark differed significantly from both leaves and roots (Tukey’s HSD, p < 0.001), whereas saponin concentrations in leaves and roots were not significantly different from each other (p = 0.065; CLD: a, b, a for leaves, bark, and roots, respectively). This preferential accumulation may reflect the involvement of saponins in protecting structural tissues against microbial invasion and insect predation, particularly in the outer protective layers of the plant (Sparg et al., 2004). Moreover, the concentration of saponins is greater in B. aristata than in B. microphylla (36.0 mg DE/g) and B. vulgaris (3.0 mg DE/g) (Furrianca et al., 2017; El-Sayed et al., 2011). From a medicinal perspective, saponins from Berberis species have shown antimicrobial, anti-inflammatory, and antioxidant activities in various studies, which is consistent with how these plants have been used traditionally across South Asia and the Middle East for centuries (Srivastava et al., 2013b). Some researchers have also pointed toward possible hepatoprotective and immunomodulatory effects, which is particularly relevant today given the growing interest in plant-based alternatives or complements to synthetic drugs (Sharma et al., 2024a). Beyond medicine, the foaming and emulsifying behaviour of saponins has also attracted attention in the food and cosmetic industries (Francis et al., 2002).

 

Total flavonoid content

Flavonoids are one of the most studied groups of secondary metabolites in higher plants because they are the main constituents of plant pigments (Awal et al., 2025). Flavonoids are potent antioxidants because of their free radical scavenging activity (Pal et al., 2009). Flavonoid content varied significantly among plant parts (F= 3678.70, p< 0.001; Table 4) As shown in Figure 5, the highest level of total flavonoid was found in leaves (54.95 ± 1.2 mg QE/g) and then showed a decreasing trend in the root (35.27 ± 0.6 mg QE/g) and stem bark (9.69 ± 0.3 mg QE/g). All pairwise comparisons were statistically significant (Tukey’s HSD, p < 0.001; CLD: c, a, b for leaves, bark, and roots, respectively). This pattern is in agreement with the physiological role of flavonoids in photoprotection, UV filtration, and antioxidant defence in photosynthetically active tissues (Fernández-Poyatos et al., 2019a). Comparable leaf-dominant flavonoid accumulation has been reported in Berberis and other medicinal taxa (Gul et al., 2023).

Total phenol content

Total phenol content differed significantly among plant parts (F= 3608.71, p < 0.001; Table 4, The total phenolic content varied from 4.23 to 62.23 mg GAE/g. Leaf extract of B. aristata contained the highest phenolic content, followed by roots. Stem bark of B. aristata contained the least amount of phenol (4.23 ± 0.9 mg GAE/g) (Figure 6). While comparing different plant parts of Berberis species, greater amounts of phenols and flavonoids in leaves of B. vulgaris and B. croatica were reported (Koncic et al., 2010). Higher phenolic quantity in leaves is expected due to their function as major antioxidants and photoprotective compounds in metabolically active aerial tissues (Fernández-Poyatos et al., 2019a). In line with these observations (Zhaleh et al., 2025), Berberis integerrima has also been shown to preferentially accumulate phenolics in its leaves compared to roots and stems.

 

Conclusion

In summary, the occurrence patterns of alkaloids, saponins and phenolic compounds in various organs of B. aristata show differences and there are certain organ-preference in these compounds. It has been shown that roots have the highest level of alkaloids. The dominant compounds in the leaf were total flavonoids, tannin and phenol while saponins occurred in the highest concentration in the stem bark. The findings suggest that aerial organs including the leaf and stem bark which are usually not much utilized could be good substitutes for roots in the extraction of specific compounds. The information is important in the view of sustainable use and conservation of the species as B. aristata have been considered to be critically endangered in the face of overexploitation of roots. In conclusion, the research makes a contribution towards improving our knowledge about B. aristata in terms of its phytochemistry and helps pave the way for alternative exploitation of certain organs for medicinal or industrial purposes. Further research is necessary on compound identification and bioactivity validation.

Acknowledgments

This work was supported by the Centre of Excellence in Herbal Technology, Forest Products and Utilization (FPU), Faculty of Forestry (FOF), Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, which provided the necessary laboratory facilities and technical support.

Novelty Statement

The present study bridges a critical knowledge gap by providing an integrated phytochemical and phsicochemical assessment of different plant parts of Berberis aristata using multiple extraction solvents. The findings establish leaves and stem bark as valuable reservoirs of bioactive compounds, offering a sustainable substitute for root exploitation. This work not only advances the phytochemical understanding of an endangered Himalayan medicinal plant but also provides a scientific foundation for its conservation and sustainable utilization in the pharmaceutical and herbal industries

Author’s Contribution

Nisha Tariq: Conceptualization, field investigation, data collection, data analysis, and manuscript writing (original draft preparation).

A.R. Malik: Supervision, study design, project administration, and critical review and editing of the manuscript.

Nazir A. Pala: Assistance in data interpretation and manuscript editing.

Peerzada Ishtiyak Ahmad: Provision of laboratory facilities and technical support.

P.A. Sofi: Management and coordination responsibility for the research activity planning and execution.

Tahir Mushtaq: Visualization and data presentation.

Amerjeet Singh: Verification of the overall replication, reproducibility of results, and other research outputs.

Mehvish Mushtaq and Moamenla Jamir: Manuscript review and editing.

Peerzada Tabish Fayaz: Providing technical assistance during laboratory work.

Generative AI and AI assisted technology statement

The authors declare that no generative AI and AI assisted technology was used in the creation of this manuscript.

Conflict of interest

The authors have declared no conflict of interest.

References

Alamzeb, M., Khan, M.R., Ali, S., Shah, S.Q. and Rashid, M.U., 2013. Antimicrobial properties of extracts and compounds isolated from Berberis jaeschkeana. Bangladesh J. Pharmacol., 8(2): 107-109. https://doi.org/10.3329/bjp.v8i2.13551

Ali, M., Malik, A.R. and Sharma, K.R., 2008. Vegetative propagation of Berberis aristata DC. an endangered Himalayan shrub. J. Med. Plants Res., 2(12): 374-377.

Alvarez, M.A., Eraso, N.F., Pitta-Alvarez, S.I. and Marconi, P.L., 2009. Two-stage culture for producing berberine by cell suspension and shoot cultures of Berberis buxifolia Lam. Biotechnol. Lett., 31(3): 457-463. https://doi.org/10.1007/s10529-008-9875-2

Awal, R., Pant, D.R. and Joshi, G.P., 2025. Impact of elevation and soil characteristics on phytochemical constituents and antioxidant potential of Berberis aristata and Berberis asiatica. Banko Janakari, 35(1): 12-21. https://doi.org/10.3126/banko.v35i1.64913

Bhatt, I.D., Rawat, S., Rawal, R.S. and Dhar, U., 2012. Antioxidant properties of Berberis asiatica roots and its extracts. J. Food Composit. Anal., 25(2): 220–224.

Chauhan, N.S., Sharma, V. and Thakur, M., 2014. Phytochemical screening and moisture content analysis of selected Berberis species. Int. J. Pharma. Sci. Rev. Res., 27(2): 150–154.

Choudhary, S., Kaurav, H., Madhusudan, S., and Chaudhary, G., 2021. Daruharidra (Berberis aristata): Review based upon its Ayurvedic properties. International Journal for Research in Applied Sciences and Biotechnology., 8(2): 98-106. https://doi.org/10.31033/ijrasb.8.2.12

El-Sayed, M., Ghareeb, D., Sarhan, E. and Khalil, A., 2011. Therapeutic bio-screening of the bioactive ingredients of Berberis vulgaris. Funct. Plant Sci. Biotechnol., 5(1): 63-68.

Fernández-Poyatos, M.D.P., Ruiz-Medina, A., Zengin, G. and Llorent-Martínez, E.J., 2019a. Phenolic characterization, antioxidant activity, and enzyme inhibitory properties of Berberis thunbergii leaves. Molecules, 24(22): 4171. https://doi.org/10.3390/molecules24224171

Francis, G., Kerem, Z., Makkar, H.P.S. and Becker, K., 2002. The biological action of saponins in animal systems: A review. Br. J. Nutr., 88(6): 587–605. https://doi.org/10.1079/BJN2002725

Furrianca, M.C., Alvear, M., Zambrano, T., Fajardo, V. and Salazar, L.A., 2017. Phytochemical constituents of the root of Berberis microphylla. Asian J. Pharma. Clin. Res., 10(6): 225-227. https://doi.org/10.22159/ajpcr.2017.v10i6.17803

Garhwal, S., Singh, R. and Bansal, V., 2010. Estimation of berberine content in different species of Berberis by HPLC. J. Med. Plants Res., 4(23): 2490–2493.

Government of India. (2007). The Ayurvedic Pharmacopoeia of India (Part I, Vol. VI). Ministry of Health and Family Welfare, Department of AYUSH. Controller of Publications, New Delhi.

Gul, Z., Akbar, A., Naseem, M., Achakzai, J.K., Rehman, Z.U. and Khan, N.A., 2023. Phytonutrient and antinutrient components profiling of Berberis baluchistanica Ahrendt bark and leaves. J. King Saud Univ. Sci., 35(2): 102517. https://doi.org/10.1016/j.jksus.2022.102517

Harborne, J.B. and Williams, C.A., 1973. A chemotaxonomic survey of flavonoids and simple phenols in leaves of the Ericaceae. Botan. J. Linn. Soc., 66(1): 37-54. https://doi.org/10.1111/j.1095-8339.1973.tb02159.x

Imenshahidi, M. and Hosseinzadeh, H., 2016. BerSberis vulgaris and berberine: An update review. Phytother. Res., 30(11): 1745–1764. https://doi.org/10.1002/ptr.5693

Katiyar, S.K., Singh, R.K. and Ali, M., 2012. Berberis aristata: A review of its pharmacological activities and phytochemistry. Int. J. Pharma. Sci. Res., 3(12): 4563–4572.

Kaur, R., Urvashi and Kumar, A., 2023. Species of the Berberis genus found in the western Himalayas. In: (eds. Sharma, A., Nayik, G.A.) Immunity Boosting Medicinal Plants of the Western Himalayas pp. 107-143 Springer, Singapore. https://doi.org/10.1007/978-981-19-9501-9_5

Khan, I., Najeebullah, S., Ali, M. and Shinwari, Z.K., 2016. Phytopharmacological and ethnomedicinal uses of the Genus Berberis (Berberidaceae): A review. Trop. J. Pharma. Res., 15(9): 2047-2057. https://doi.org/10.4314/tjpr.v15i9.33

Komal, S., Ranjan, B., Neelam, C., Birendra, S. and Kumar, S.N., 2011a. Berberis aristata: A review. Int. J. Res. Ayurveda Pharmacol., 2(2): 383-8.

Končić, M.Z., Kremer, D., Karlović, K. and Kosalec, I., 2010. Evaluation of antioxidant activities and phenolic content of Berberis vulgaris L. and Berberis croatica Horvat. Food Chem. Toxicol., 48(8–9): 2176–2180. https://doi.org/10.1016/j.fct.2010.05.025

Kumar, N., Bhandari, P., Singh, B., Gupta, A.P. and Kaul, V.K., 2008. Reversed phase-HPLC for rapid determination of polyphenols in flowers of rose species. J. Separat. Sci., 31(2): 262-267. https://doi.org/10.1002/jssc.200700372

Kumar, S. and Pandey, A.K., 2013. Chemistry and biological activities of flavonoids: An overview. Sci. World J., pp. 162750. https://doi.org/10.1155/2013/162750

Malik, A.R., Tariq, N., Sofi, P.A., Singh, A., Peerzada, I.A., Mushtaq, T. and Ayoob, B., 2024a. Berberis aristata: An Endangered High Value Medicinal Shrub of Western Himalayas in Peril. Adv. Concepts Pharma. Res., 9: 1-15. https://doi.org/10.9734/bpi/acpr/v9/3172G

Mipun P., Rio, P., Nabanita, H., Hiranjyoti, D., Amenuo, S.K., Puranjay, M., Petekhrienuo, R., Mayor, M.P. and Debajit, K., 2023. Phytochemicals in medicinal plants: Biodiversity, bioactivity and drug discovery, pp. 1

Moin, M.S., Siddiqui, J.I., Khan, S., Alam, M.A., Khatoon, F. and Alam, A., 2023. Therapeutic potential and traditional values of frequently used medicinal plant Berberis aristate DC. (Rasaut): A comprehensive review. Res. J. Pharma. Technol., 16(4): 2035-2043. https://doi.org/10.52711/0974-360X.2023.00335

Ncube, N.S., Afolayan, A.J. and Okoh, A.I., 2008. Assessment techniques of antimicrobial properties of natural compounds of plant origin: Current methods and future trends. Afr. J. Biotechnol., 7(12): 1797–1806. https://doi.org/10.5897/AJB07.613

Nisha, T., 2023. Population assessment and phytochemical characterization of Berberis aristata DC. in Kashmir Himalayas. M. Sc. thesis, Faculty of Forestry, SKUAST-K, Benhama Ganderbal, India.

Obadoni, B.O. and Ochuko, P.O., 2002. Phytochemical studies and comparative efficacy of the crude extracts of some haemostatic plants in Edo and Delta States of Nigeria. Glob. J. Pure Appl. Sci., 8(2): 203-208. https://doi.org/10.4314/gjpas.v8i2.16033

Och, A. and Nowak, R., 2021. Barberry (Berberis vulgaris) traditional and contemporary use. In: Ekiert, H.M., Ramawat, K.G., Arora, J. (eds), Medicinal Plants: Sustainable Development and Biodiversity, vol. 28, 1st edn. pp. 797-825. Springer, Cham., https://doi.org/10.1007/978-3-030-74779-4_24

Pal, D., Sannigrahi, S. and Mazumder, U.K., 2009. Analgesic and anticonvulsant effects saponin isolated from the leaves of Clerodendrum infortunatum Linn. in mice. Indian J. Exper. Biol., 47(9): 743-747. https://doi.org/10.3742/OPEM.2009.9.2.128

Pharmacopoeia, I., 1996. Ministry of health and family welfare. Government of India, 2: 350.

Pizzi, A. and Cameron, F.A., 1986. Flavonoid tannins structural wood components for drought-resistance mechanisms of plants. Wood Sci. Technol., 20(2): 119-124. https://doi.org/10.1007/BF00351023

Pizzi, A. (2021). Tannins: Medical/pharmacological and related applications—A critical review. Sustainable Chemistry and Pharmacy, 22, 100481

Sharma, S., Gupta, S., Dhar, M. K., & Kaul, S. (2018). Diversity and Bioactive Potential of Culturable Fungal Endophytes of Medicinal Shrub Berberis aristata DC.: A First Report. Mycobiology, 46(4), 370–381.

Sharma, S., Chaitanya, M.V.N.L., Sharma, S., Kumar, S., Rustagi, S., Singh, S. and Yadav, A.N., 2024. The medicinal plant Berberis aristata and its endophytes for pharmacological applications: Current research and future challenges. J. Appl. Biol. Biotechnol., 12: 37-46. https://doi.org/10.7324/JABB.2024.167591

Singleton, V.L. and Rossi, J.A., 1965. Colorimetry of total phenolics with phosphomolybdic–phosphotungstic acid reagents. Am. J. Enol. Viticult., 16(3): 144–158. https://doi.org/10.5344/ajev.1965.16.3.144

Sofowara, A., 1993. Recent trends in research into African medicinal plants. J. Ethnopharmacol., 38(2-3): 197-208. https://doi.org/10.1016/0378-8741(93)90017-Y

Sparg, S.G., Light, M.E. and Van Staden, J., 2004. Biological activities and distribution of plant saponins. J. Ethnopharmacol., 94(2): 219–243. https://doi.org/10.1016/j.jep.2004.05.016

Srivastava, S., Srivastava, M. and Rawat, A.K.S., 2013b. Comparative phytochemical and antioxidant analysis of different Berberis species. J. Med. Plants Res., 7(15): 1024–1030.

Sutalangka, C., Wattanathorn, J., Muchimapura, S. and Thukham-mee, W., 2013. Screening of antioxidant activity and phytochemical constituents of Berberis species. J. Med. Plants Res., 7(9): 556–563. https://doi.org/10.1155/2013/695936

Thakur, M., Sharma, P. and Anand, A., 2019. Seed priming-induced early vigor in crops: An alternate strategy for abiotic stress tolerance. In: Hasanuzzaman, M., Fotopoulos, V. (eds), Priming and Pretreatment of Seeds and Seedlings: Implication in plant stress tolerance and enhancing productivity in crop plants, 1st edn. pp. 163-180. Springer, Singapore. https://doi.org/10.1007/978-981-13-8625-1_8

Van Buren, J.P. and Way, R.O., 1981. Tannin hazes in deproteinized apple juice. J. Food Sci., 43(4): 1235-1237. https://doi.org/10.1111/j.1365-2621.1978.tb15276.x

Verma, N., Singh, J.P., Mishra, D., Nath, A., Kashyap, P. and Dutta, D., 2017. Indian barberry (Berberis aristata DC). In: Ghosh, S.N., Singh, A., Thakur, A. (eds), Underutilized Fruit Crops: Importance and Cultivation, 1st edn. pp. 143-154. Jaya Publishing House, New Delhi, India.

World Health Organization, 1998. Quality control methods for medicinal plant materials. World Health Organization.

Yadav, R., 2018. The pharmacognostical studies of Berberis aristata DC. International Journal of Green Pharmacy, 12(2).

Zhaleh, N., Sharifi, M., Samari, E., Halvagar, M.R. and Zeidi, S.H., 2025. A comparative study of alkaloid and phenolic compounds in different organs and tissues of Berberis integerrima. PLoS One, 20(5). https://doi.org/10.1371/journal.pone.0321255