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  • Scientific DSR Validation, Pharmacognostical, Biodiversity, Toxicological Research Studies of Nyctanthes Arbortristis L.- Aerial Leaves Part and Their Pharmacological, Therapeutic Medicinal Values

  • Photo Dr.Pawan Kumar Sagar* Photo Sajwan S. Photo Sri P. M. D. Photo Kashyap S. Photo Venkatesan K. Photo Jayanthy A. Photo Kumar M.

  • 1,2,4 Drug Standardization Research Institute, (Under CCRUM, Ministry of AYUSH., Govt. of India), PCIM&H Campus, IInd Floor, Kamla Nehru Nagar, Ghaziabad, U.P., India.
    3,5Regional Research Institute of Unani Medicine, (Under CCRUM, Ministry of AYUSH) Govt. of India, Royapuram, Chennai, TN, India.
    6,7Pharmacopoeia Commission for Indian Medicine & Homoeopathy (PCIM&H), Kamla Nehru Nagar, Ghaziabad UP. India (Ministry of AYUSH, Govt. of India)

Abstract

Botanical, Pharmagconistical, Biodiversity and Toxicological research studies of ASU herbal products remains a big challenging task on global levels. There needs to be more than the advance investigation research studies and screening parameters to validation, authenticate, identification and differentiate adulterants.. Nyctanthes arbortristis L. is one of the herbs used to treat various health wellness and therapeutic illness of public mankind from since ancient time. This study aims to evaluate the Botanical, Pharmacognostical, Biodiversity and Toxicological research studies of the plant of NAT. The Botanical, Pharmacognostical, Biodiversity and Toxicological authenticate identification, quality control research studies of the plant of NAT powder were carried out using standard methods. The studies explored of quality, safety and toxicity effects of the tested drug samples were also investigated applied standard methods WHO/AOAC/AYUSH pharmacopeias. The Botanical, Pharmacognostical, Biodiversity, Toxicological QC. QA. Properties of NAT have shown that all the investigated parameters were within the permissible limits. The tested drug samples showed significant quality, safety and toxicity studies against certain pathogens organisms and promising anti-pathogenic activity. In the investigated studies of Botanical, Pharmacognostical, QC. Toxicological research findings revealed that the revalidated test drug was free from adulterations and toxic contaminations. This investigated herb research data confirmed to revalidated of drug developed standard Atlas, Pharmacovigilance and therapeutic medicinal values may treat that the drug is safe for internally use and cures in as Antiarthritis, Antistress, Anti-inflammatory, Antimicrobial, Antibacterial, Antifungal, Hypoglycemic and hypolipidemic activity, Antiviral activity, Antiulcer activity, Analgesic activity, Anticancer activities, Cytotoxic activities, Obstinate Sciatica disorder etc.

Keywords

Nyctanthes arbortristis L. Botanical, Pharmacognostical, Biodiversity, QC. and QA, Toxicological, Pharmacological Quality and Safety studies, therapeutic medicinal values.

Introduction

The World Health Organization (WHO) reports that herbal medicines are able to fulfill the health requirements of approximately 80% of the global population, particularly those residing in rural regions of developing nations. The quality assurance and quality control of herbal crude drugs and formulated products are important in justifying their acceptability in modern system of medicine. Hence it is required to conduct the advance research on drugs standardization and product validation to provide effective, curable and safe drugs to the needy mass suffering from various ailments.(1-2),(9-15),(19-20),(27) As per Hindu mythology, the Harsinghar/ Parijata tree is recognized as one of the five wish-granting trees associated with Devaloka.(6),(17)  N. arbor-tristis is commonly referred to as “Night Jasmine or Harsinghar.” The genus name “Nyctanthes” is derived from the Greek phrases “Nykhta,” meaning night, and “anther,” meaning flower. Within the botanical family of Oleaceae, Night Jasmine genus encompasses more than 600 species of little trees and vines.(6),(17) A species of plant known as N. arbortristis and these glabrous twining shrubs are commonly found in tropical Asia and warm temperate regions of Europe and Africa. Additionally, they are extensively cultivated in gardens within these areas. [6],[82] The plant is widely recognized across the country for its aromatic white flowers. The plant is distributed over several regions, namely southern India, northern Pakistan, Thailand, Malaysia, and Indonesia. Its original habitat is the subtropical Himalayas of Nepal and India. (6),(82) The species is distributed over several regions in India, including the outer Himalayas, the Jammu and Kashmir region, East Assam, Bengal, Tripura, and the central region extending to the Godavari River in the southern part. The plant flourishes in soil that is reddish-black in color and has a pH level between 5.6 and 7.5. It is well-suited for dry and semi-arid conditions.[6],[8],[26] At present, there is a desperate need for the development of quality parameters for herbal drugs and raw plant materials. The WHO has emphasized the need to ensure the quality of medicinal plant products by using any advanced controlled techniques. (118),(121)          

Nomenclature:

Sanskrit:

Parijatha/ Sephalika, Rajanikasa

Marath:

Parijathak/ Khurasli/ Partaka

Hindi:

Harsingar / Siharu

Gujarathi:

Jayaparvati

English:

Night Jasmine/ Coral jasmine/ Weeping nyctanthes

Telugu:

Pagadamalle/ Shwetasurasa

Bengali:

Sephalika / Singhar

Oriya:

Gangasiuli

Malayalam:

Parijatakam/ Manpumaram

Kannada:

Parijatha.

Punjabi:

Kuri/Laduri

Tamil:

Pavalamalligai

Urdu:

Harsingar

 

 

Taxonomical Classification:

 

Kingdom:

Plantae

Divisin:

Magnoliophyta

Class:

Magnoliopsida

Order:

Lamiales

Family:

Oleaceae

Genus:

Nyctanthes

Species:

Arbortristis

Binomial Name:

Nyctanthes arbortristis Linn.

Bio diversity of herbaceous medicinal plant Nyctanthes arbortristis L.:

 

Name of plant

 

Worldwide Biodiversity and introduced

from native regions

References

Nyctanthes arbortristis L.

 

 

 

Commonly found in tropical Asia and warm temperate regions of Europe and Africa, distributed over several regions, namely southern India, northern Pakistan, Thailand, Malaysia, and Indonesia. distributed over several regions in India, including the outer Himalayas, the Jammu and Kashmir region, Andhra Pradesh, East Assam, Bengal, Tripura, Orissa, Burma, Central India like Chhatanagpur, Rajasthan, Madhya Pradesh and southwards to Godavari and the central region extending to the Godavari River in the southern part and Asian region of Nepal and Sri Lanka.

 

Kaliyaperumal et al., 2024(6); Panda et al., 2024(8); Rawat et al., 2021(26); Acharya, 2011 (82); Sasmal et al.,2007 (95); Kirtikar and Basu, 2000 (117); Anonymous, Wealth of India,1997 (118) ; Varier,1995(124); Kiew and Bass, 1984 (135) ; Nadkarni,1982 (136)

NAT Investigated herbasious medicinal plant Whole plant parts Dried Stem barks, Leaves, Flower, Fruits with seeds parts , Dried Seeds and their investigated studies Graphical Illustration, investigated plant Confirmation and identification by Herbarium sheets shown in Fig.-1, Fig.-2, Fig.-3, and Fig.-4  respectively.

       
            Fig.-1, Graphical Illustration.png
       
Fig.-1, Graphical Illustration

       
            Botanical authentication of  N. arbortristis by Authentic Specimen Herbarium Sheet basis.png
       
Fig.-2, Botanical authentication of  N. arbortristis by Authentic Specimen Herbarium Sheet basis.

       
            Dried Steam Bark. Leaves, Flowers part of N. arbortristis.png
       
Fig.-3, Dried Steam Bark. Leaves, Flowers part of N. arbortristis

       
            Fresh and Dried Seeds part of N. arbortristis.png
       
Fig.-4, Fresh and Dried Seeds part of N. arbortristis

MATERIAL AND METHODS:

Drug Collection and authentication, plant collection source: (1-2),(9-13),(19-20),(24),(41),(104-106),(116),(118),(126-127)    

The raw drugs samples were procured and collected of the plant parts of NAT from the Central North - West region, Herbal Garden, PCIM&H Campus, Pocket-II, Scientific Block, Kamla Nehru Nagar, Ghaziabad UP. India, South - North  Region, RRIUM, Herbal Garden, Tamil Nadu State, India and North western Himalayas -  Region, Shivalik Nagar, L-Block, Haridwar, Uttarakhand State, India and investigated plant parts of NAT samples authenticated and confirmed by Botany and Pharmacognosy Laboratory Section, Researcher Scientific Staff of Regional Research Institute of Unani Medicine, Royapuram, TN. State, Drug Standardization Research Institute, Botany Section, Ghaziabad, UP. and re-authenticated and reconfirmed by PCIM&H, Botany Department ,Ghaziabad UP. India by the Authenticated confirmation of Herbariums Specimen Sheet basis. Researcher staffs using phamacognostical standard methods after proper authentication, plant parts of NAT samples were washed thoroughly with clean water and dried under a gentle steam of air in the laboratory till no loss in weight temperature 30°C and powdered in an electric grinder.

Botanical and Pharmacognostical Studies: (1-2),(9-13),(19-20),(24),(116-117),(126-127) The dried Arial part of NAT subjected to macroscopic studies as per approved format of API/UPI,AYUSH India standard methods and evaluated systematically. Thin transverse section were taken from seeds samples stained with safranin and mounted in glycerin by following the micro technique methods. Microphotography was performed for the drug.(13),(104),(126) (127),(131) Quantitative Microscopy: The cleared materials were washed thoroughly and stained with safranin for quantitative microscopic studies. Maceration Study: Shade dried and coarsely powdered plant was treated with Jeffrey’s reagent for a few hours. The action of the macerating fluid was stopped before the complete separation of all cells. Then the macerated tissue was carefully washed in distilled water to remove as much of the acid as possible and then transferred to 50% alcohol for study. Slides were made by placing small quantities of cells in water on a slide. The excess water was evaporated, mounted in glycerin and observed through microscope.

Toxicological investigation study: (1-2),(9-15),(19-20),(22-23),(24),(104-106),(118),(121), (126-128),(131),(134) Standard Methods applied for detection and investigation of Toxicology study parameters, WHO/AOAC/AYUSH. Pharmacopeial permissible standard Limits basis, All investigated and tested samples of NAT have been carried out as per WHO/AOAC/AYUSH-API/UPI Standard methods. Botanical, Pharmacognostical powder microscopical analysis, authentication and identification of fresh and dried Arial parts NAT.:

       
            FIG-5.png
       
    
       
            Nyctanthes arbortritis Linn..png
       
Fig.-5, Nyctanthes arbortritis Linn. Aerial portion Microscopy

       
            Powder Microscopy.png
       
Fig.-6, Powder Microscopy

        
            FIG-7.png
       

RESULT AND DISCUSSION: 

Macroscopic investigation: Investigated drug NAT occurred in pieces of varying thickness ranging from 0.4 to 0.6cm diameter and upto 7cm length; young stems green with smooth surfaces and swelling at nodes, older ones show a light brown; leaves simple, petiolate, petiole very small upto 0.7cm; lamina ovate with acute leaf tip, 3 to 12cm in length and 1.5 to 8cm in breadth, margin entire to slightly serrated, venation unicostate reticulate with pinnate incision; leaves dorsiventral, upper surface deep green in colour, slightly rough, hairs or trichomes very fine but firm (hirsute), lower surface light green in colour, smooth and soft; odour herbaceous, taste bitter and slightly astringent.

Microscopic investigation: Investigated drug NAT occurred in Arial stem part : T.S. of stem shown anomalous secondary structure; circular in outline with quadrangular ridges; presence of cortical vascular bundles apart from normal vascular bundle; epidermis consisting of single layer of thick walled parenchyma cells covered with cuticle; trichomes numerous, long thick walled cells with tapering ends, cortex consisting of few layers of collenchyma, chlorenchyma and parenchyma cells; pericycle consisting of patches of few sclerenchyma fibres;  normal vascular bundles ring shaped occur in the central region with xylem elements towards the centre and phloem toward outside; four inversely oriented cortical vascular bundles present at four ridges with xylem towards outside and phloem towards inside;  the vascular bundles collateral and open;  pith present in the centre. Petiole: T.S. of petiole shown prominent notch towards the middle of upper peripheral zone; epidermis consisting of single layer of thick walled parenchyma cells covered with cuticle; trichomes numerous long thick walled tapering cell; cortex consisting of few layers of collenchyma, chlorenchyma and parenchyma cells; vascular bundles 3 to 7 in number, bigger vascular bundles in the centre, where as lateral bundles comparatively smaller in size; each bundle consists of xylem towards the upper side and phloem towards the lower side of the bundle; a few sclerenchyma fibres present on the lower side of the vascular bundle. Leaf through Midrib: T. S. of leaf through midrib shows epidermis consisting of single layer of thick walled parenchyma cells with cuticle, numerous unicellular trichomes present on both the upper and lower side; cortex consisting of few layers of collenchyma, chlorenchyma and parenchyma cells; vascular bundle sickle shaped in the centre with xylem towards the upper side and phloem towards the lower side of the bundle; a few sclerenchyma fibres on the lower side of the vascular bundle. Lamina: T.S. of lamina shown dorsiventral; epidermis consisting of single layer of thick walled parenchyma cells with cuticle; trichomes of variable sizes present on both the upper and lower epidermis; palisade parenchyma consisting of two layers of elongated parenchyma cells on the upper side followed by 8 to 9 layers of spongy parenchyma on the lower side; numerous scattered stomata present on the lower epidermis; epidermal cells in surface view consisting of polygonal parenchyma cells with angular walls; anisocytic stomata present only in the lower epidermis; stomatal number of the lower epidermis 50 to 55/sq mm and stomatal index of the lower epidermis 40 to 42/sq mm; palisade ratio 9 to 11 and vein islet number 14 to 19. Shown in Fig.-5 respectively.   

Powder Microscopy: Grayish brown; numerous elongated thick walled mostly unicellular trichomes upto 400? with gradually tapering tips; vessels with pitted thickening up to 45µ; thick walled elongated cells of fibers and tracheids up to 1300µ and width upto 35µ; xylem parenchyma cells; upper epidermal cells in surface view with only trichomes; lower epidermal cells angular walls in surface view with anisocytic stomata and trichomes; palisade parenchyma cells in double layers and parenchyma cells in surface view. Shown in Fig.-6 respectively.   

Ethno-medicinal uses and Biodiversity of investigated plant NAT: (4),(6),(8),(18),(26),(82),(95),(117),(121),(122),(133-134)

  The investigated herbaceous medicinal plant NAT was worldwide, ethno-medicinal uses and biodiversity occurred and found in tropical Asia and warm temperate regions of Europe and Africa, distributed over several regions, namely southern India, northern Pakistan, Thailand, Malaysia, and Indonesia. distributed over several regions in India, including the outer Himalayas, the Jammu and Kashmir region, Andhra Pradesh, East Assam, Bengal, Tripura, Orissa, Burma, Central India like Chhatanagpur, Rajasthan, Madhya Pradesh and southwards to Godavari and the central region extending to the Godavari River in the southern part and Asian region of Nepal and Sri Lanka uses by these Tribal’s population’s which have lived in these forest region from since ancient time that was found in the study plant used by Tribal’s population’s very broad level’s, already gained of knowledge, well aware and well known upon practically therapeutic uses of study plant of NAT and well aware about their medicinal efficacy and medicinal values,  wealthiest aspects and curable potent properties as Anti-inflammatory, Anti-helmintic, Antibilious, Antibacterial, Antifungal activities and Analgesic activity, Cold, Joint and Sciatica pain potential, illness, disorder’s for curable of our health and these shown have beneficial medicinal curing potential developed of NAT from since ancient time. The study plant NAT had occurred ethno botanically, climatically and biodiversity appeared world wide occurrences presence in rich form of investigated herbaceous medicinal plant. Investigated medicinal plant NAT has been present and confirmed bioactive phytochemical constituents having numerous secondary metabolites, detail shown in Table-1 respectively.    

Toxicological investigated study : (1-2),(9-15),(19-20),(22-23),(24),(28-29),(41),(46),(104-1061),(118),(131) Studies Medicinal plant NAT has been using and applied of Advance sophisticated Instruments and operating parameters detections and investigations of Heavy Metals - Pb, As, Cd, Hg in ppm. levels by Thermo Fisher M Series, 650902 V1.27 model Atomic Absorption Spectrometer with Graphite Furnace (AAS-GF), Aflatoxins - B1.B2 and G1,G2 in ppm. levels were estimated by Kobra cell techniques using CAMAG or Anchrom HPTLC instrument, Revolution Front (Rf) values detect in cm. range, Detector - UV-Visible detector, Detector temperature :25°C, Sample injector volume range - 5.0µl to 20µl range, Pesticide residues in mg/kg and bio active components were  analyzed using Gas Chromatography Mass Spectra (GC-MS) (Instrument- Thermo Scientific, Model TSQ9000), detector-mass selective detector or Triple Quadrupole mass analyzer detector, column specification-TG-5MS /  Run time- usually for the investigated compounds vary according to the method of GC and temperature programming whereas roughly all the  all the  relevant bio active phyto-chemical constituents usually appear in an around 0-50 minutes in the GC column i.e. the Retention time, Run time usually varies with the method and GC temperature program. Mass value range - 0.0 to 650amu, Sample injector volume range - 1.0 µl to 5.0 µl. Microbial Load contamination - TBC/TFC detect in cfu/gm. Escherichia coli, Salmonella typhai Spp. Staphylococcus aurous author pathogenic, detection and estimation in Heavy Metals-Pb, Hg, Cd and As, Aflatoxins- B1.B2 and G1, G2 and  Pesticide Residues - Organo chlorine, pesticides, Organo phosphorus pesticides, Pyrethroids etc, in ppm levels concentrations. NAT. having investigated bioactive phytochemical constituents with immense, remarkable therapeutic medicinal potential values, pharmacological action properties, shown in Table-1 respectively, and Toxicologically investigated of collected samples of various 3 regions from India, arial plant part 3 samples of NAT. medicinal plant shown, toxicological QC., QA. research data’s. Toxicology investigated research parameters revealed results were shown with in prescribed WHO/AYUSH Pharmacopeial permissible standard Limits in as Microbial Load contaminations - TBC/TFC detect in cfu/gm. Escherichia coli , Salmonella typhai Spp. Staphylococcus aurous author pathogenic, detection and in Heavy Metals-Pb, Hg, Cd and As, Aflatoxins B1,B2 and G1,G2 and  Pesticide Residues - Organo chlorine, pesticides, Organo phosphorus pesticides, Pyrethroids etc. in ppm. Level’s and potentially toxic elements detections in ppb. levels. Resulted the all investigated Toxicology research parameters found in the study medicinal plant investigated 3 samples shown complies and not found of any hazardous or highly toxic contamination in the investigated drug samples, It had fit for internal used. Investigated results shown in Table’s - 4,5,6 and 7 respectively.

 

Table -1, Investigated Major Active- Phyto-chemical Constituents in NAT:

 

Plant

 

Part Used

Major Active- Phytochemical Constituents

References

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Nyctanthes arbortristis Linn. (Night Jasmine)

 

 

Leaves

Hexadecanoic acid (26.4%), phytol (13.6%), Hexenyl benzoate (11%), Linalool (11.3%), Octadecanoic acid (6.2%), Methyl salicylate (5.6%), n-Dodecanol (5.5%), Alpha-Terpineol (4.7%), and Geraniol (3.7%), D-mannitol, ?-sitosterole, astragaline, nicotiflorin, oleanolic acid, nyctanthic acid, tannic acid, ascorbic acid, methyl salicylate, carotene, friedeline, lupeol, mannitol, glucose and fructose, iridoid glycosides, Flavanol glycosides, benzoic acid, derivative of kaempferol, carotene.

Swain et al.,2024(4)   ;Kaliyaperumal et al.,2024(6)   ; Kushwah et al., 2023 (18)   ; Satyal et al., 2012(80)   ; Hukkeri , et al., 2006(107)   .

Stem Barks

Hexadecanoic acid (34.3%), Alpha-Eudesmol (8.7%), Beta-Eudesmol (17.1%), Elemol (5.8%), Cryptomeridiol (4.8%), Octadecanoic acid (3.9%), %), n-Dodecanol (6.8%), Methyl palmitate (1.8%), Methyl stearate (1.4%), and 1,8-Cineole (1.3%),Glycoside-naringenin-4?-0-?-glucapyranosyl-?-xylo pyranoside and ?-sitosterol, Glycosides and alkaloids.

Swain et al.,2024(4)   ;Kaliyaperumal et al., 2024 (6)   ;Kushwah et al., 2023 (18)   ;  Satyal et al., 2012 (80)   ;Vats et al., 2009 (90)   ; Girach et al., 1994 (124)

Flowers

1-octanol (74.81%), 2-hexadecen-1-ol, 3,7,11,15-tetra (6.8%), Bis (2-ethylhexyl) phthalate (5.88%), 2,4-cycloheptadiene-1-one, 2,6,6-trimethyl (4.23%), Hexadecanoic acid methyl ester (2.07%), and Benzoic acid (1.21%), Phytol (32.2%), Methyl palmitate (14.7%), cis-9-Tricosene (3.6%), Geranylgeraniol (2.7%), n-Nonadecene (2.2%), Phytone (1.4%), Methyl stearate (1.1%), n-Benzyl salicylate (1.1%), and Heptacosane (0.8%), Essential oil, Nyctanthin, D-mannitol, Tannin and Glucose, Carotenoid, glycosides viz ?-monogentiobioside ester of ?-crocetin (or crocin-3), ? monogentiobioside-?-D monoglucoside ester of ?-crocetin, ?-digentiobioside ester of ?-crocetin. (or crocin-1), 4- hydroxy hexahydrobenzofuran–7-one.

Swain et al.,2024(4)   ;Kaliyaperumal et al.,2024(6)   ; Kushwah et al., 2023 [18]   ; Karthick et al., 2019(33)   ; Sriwardena et al., 2014(52)   ; Thangavelu et al., 2010 (83)

Seeds

Arbortristoside A&B, Pale Yellow Brown Oil (15%),  Glycerides of linoleic oleic, Lignoceric, Stearic, Palmitic and Myristic acids, Nyctanthic acid, Nyctoside A, ? -sitosterol, 3-4 secotriterpene acid and A water soluble polysaccharide composed of D-glucose and D-mannos

Swain et al.,2024(4)   ;Kushwah et al., 2023 (18)   ; Adebajo et al.,2007(96)

Roots

?-Sitosterol and Oleanolic acid

 

 

Swain et al.,2024(4)   ;Jain et al.,2011(77)

Pharmacological and Therapeutics medicinal potent values of NAT: 

Nyctanthes arbortristis,(NAT) also known as Harsinghar / Parijata /coral jasmine/night jasmine, is a plant that has gained attention for its therapeutic properties and medicinal applications. NAT various plant parts has been investigated in Leaves, Flowers, Stem Bark, Seeds, Fruits In-vitro, In-vivo pharmacological studies and confirmation, explored their remarkable therapeutic and medicinal potential.(3),(6),(30) Respectively Shown in Table -2.  Anticancer activities has been investigated in  Leaves, Flower’s, Dried Fruit’s, Fruits, leaves, and Stem barks parts of NAT.(4-5),(6),(18),(31) Tumour necrosis factor Depleting activity has been investigated in Leave’s part of NAT.(30) Cytotoxic activities has been investigated in Flower’s part of NAT. (6),(30),(111),(115) Antiproliferative, Anticancer activities in Flower’s part of NAT. (4) and Cytotoxic activities has been investigated in Leaves and Stem, Stem, leaves, and Fruits parts of NAT. (30) Respectively Shown in Table -3. 

Table -2, Investigated Pharmacological Activities in In-vitro & In-vivo NAT studies:

 

Investigated

Plant parts

Used in Studies Extracts

Study plan

Pharmacological

Activities

References

Leaves

Ethanolic extract

In-vitro

Antibacterial activity

Kaliyaperumal et al.,2024 [6]; Gahtori et al., 2024[7]

Leaves

Aqueous extract

In-vitro

Antibacterial activity

Kaliyaperumal et al.,2024 [6]; Rani et al., 2023[17]

Leaves

Ethanolic, Methanolic, Petroleum ether, and Aqueous extracts

In vitro

Antibacterial activity

Parekh et al.,2020[30] ;

Vyas et al.,2013[65]

Leaves

Ethanolic extract

In vitro

Antibacterial activity

Parekh et al.,2020[30] ;

Show et al.,2014[54]

Leaves

Ethyl acetate extract

In-vitro

Anti-fungal activity

Kaliyaperumal et al.,2024 [6]

Leaves

Ethanolic extract

In-vitro

Antioxidant activity

Kaliyaperumal et al.,2024 [6]; Gahtori et al., 2024[7]

Leaves

Aqueous extract

In-vitro

Antioxidant activity

Kaliyaperumal et al.,2024 [6]; Rani et al., 2023[17]

Leaves

Petroleum ether extract

In-vitro

Antimicrobial activity

Kaliyaperumal et al.,2024 [6]; Pandey et al.,2016[14]

Leaves

Betulinic acid

In-vitro

Antioxidant activity

Kaliyaperumal et al.,2024 [6]; Karan et al.,2019[32]

Leaves

?-sitosterol isolated from petroleum ether extract

In-vivo

Analgesic activity

Kaliyaperumal et al.,2024 [6]; Nirmal et al.,2012[74]

Leaves

Petroleum ether extract

In-vivo

Anti-inflammatory activity

Kaliyaperumal et al.,2024 [6]; Nirmal et al.,2012[74]

Leaves

Ethanolic extract

In-vivo

Hypoglycemic and hypolipidemic activity

Kaliyaperumal et al.,2024 [6];Mousum et al.,2018[35]

Leaves

Petroleum ether extract

In-vivo

Hepatoprotective activity

Kaliyaperumal et al.,2024 [6] ;Chaudhary et al.,2018 [36]

Leaves

Methanol extract and Chloroform extract

In-vivo

Larvicidal activity

Kaliyaperumal et al.,2024 [6]; Pandey et al.,2016[42]

Leaves

Ethanolic Extract

In-vivo

Antiviral activity

Parekh et al.,2020[30]   ; Kannan et al.,2007[99]

Leaves

Aqueous Extract

In-vivo

Antiviral activity

Parekh et al.,2020[30] ;

Kannan et al.,2010[99]

Leaves

Water-soluble fraction

In-vivo

Immunostimulatory activity

Parekh et al.,2020[30];

Devasree et al.,2014 [53]

Leaves

?-sitosterol isolated from petroleum ether extract

In-vivo

Immunostimulatory activity

Kaliyaperumal et al.,2024 [6]; Parekh et al.,2020[30]; Nirmal et al.,2012[74]

Leaves

95 % ethanolic extract

In-vivo

Anti-inflammatory activity and Analgesic activity

Parekh et al.,2020[30] ;

Saxena et al.,1987[129]

Leaves

90% ethanolic extract

In-vivo

Anti-inflammatory activity and Analgesic activity

Parekh et al.,2020[30] ;

Pattanayak et al.,2013[61]

Leaves

90% ethanolic extract

In-vivo

Cognitive impairment

Parekh et al.,2020[30];

Phanindhra et al.,2015[47]

Leaves

Water-soluble extract

In-vivo

Ulcerogenic activity and Antipyretic

Parekh et al.,2020[30] ;

Saxena et al.,1987[129]

Leaves

Aqueous extract

In-vivo

Antipyretic

Parekh et al.,2020[30];

Bhatia et al.,2001[113]

Leaves

95% Ethanolic, 50% hydro-alcoholic

In-vivo

Immuno-modulator/ Immunorestorative activity

Parekh et al.,2020[30];

Agrawal et al.,2013[62]

 

Leaves

95% ethanolic extract

In-vivo

Antiarthritic activity

Parekh et al.,2020[30];

Goyal et al.,2013[64]

Leaves

Ethyl acetate extract

In-vivo

Antiarthritic activity

Parekh et al.,2020[30] ;

Uroos et al.,2017 [38]

Leaves

Methanolic extract

In-vivo

Hepato-protective activity

Parekh et al.,2020[30];

Vishwanathan et al.,2010[89]

Leaves

Methanolic extract

In-vivo

Hepato-protective activity

Parekh et al.,2020[30];

Mahida et al.,2007[102]

Leaves

Ethanolic extract

In-vivo

Hepato-protective activity

Parekh et al.,2020[30];

Sathiya et al.,2008[94]

Leaves

50% Ethanolic extract

In-vivo

Antidiabetic activity

Parekh et al.,2020[30];

Husain et al.,2010[84]

Leaves

Ursolic acid

In-vitro

Antifilarial activity

Parekh et al.,2020[30];

Saini et al.,2014[56]

Leaves

Herbal Formulation preparation (250mg powder/5 ml suspension

In- vivo

Antifilarial activity

Parekh et al.,2020[30];

Ghiware et al.,2007[98]

Leaves

Fresh paste of leaves

In-vivo

Antifilarial activity

Parekh et al.,2020[30];

Karnik et al.,2008[93]

Leaves

Ethanolic extract

In-vitro

Antifilarial activity

Parekh et al.,2020[30];

Kumari et al.,2012a[78]

Leaves

Fresh preparation of leaves paste

In-vivo

Antimalarial activity

Parekh et al.,2020[30];

Godse et al.,2016[45]

Leaves

Methanolic extract

In-vivo

Wound healing activity

Parekh et al.,2020[30];

Sopi et al.,2013[58]

Leaves

Ethanolic extract

In-vivo

Hypoglycemic activity

Kaliyaperumal et al.,2024 [6]; Mobiya et al.,2023[16]

Leaves

Aqueous extract

In-vitro

Antioxidant activity

Parekh et al.,2020[30] ;

Dasgupta et al.,2007[97]

Leaves, Stem

Ethanolic extract

In-vitro

Immuno-modulator/ Immunorestorative activity

Parekh et al.,2020[30] ;

Thomas et al.,2013[63]

Flowers

Dry flower aqueous extract

In-vitro

Antioxidant activity

Parekh et al.,2020[30] ;

Vankar et al.,2008[92]

Flowers

Ethanolic, Ethyl acetate, and Aqueous extract

In-vitro

Antioxidant activity

 

Kaliyaperumal et al.,2024 [6]; Mishra et al.,2016[43]

Flowers

Ethanolic extract

In-vitro

Antibacterial activity

Kaliyaperumal et al.,2024 [6]; Gahtori et al., 2024[7]

Flowers

Ethanolic extract

In vitro

Antibacterial activity

Parekh et al.,2020[30];

Srinivasan et al.,2011[81]

Flowers

Alcoholic extract utilized for the synthesis of silver nanoparticles

In vitro

Antibacterial activity

Parekh et al.,2020[30] ;

Gogoi et al.,2015[18]

Flowers

Water-soluble fraction of 70% ethanolic extract

In-vivo

Antiarthritic activity

Parekh et al.,2020[30];

Deshmukh et al.,2007[101]

Flowers

Aqueous extract

In-vitro

Anti-fungal activity

Kaliyaperumal et al.,2024 [6] ; Jamdagni et al.,2018[37]

Flowers

Crude aqueous extracts -Methanol fraction

Hexane fraction

In-vitro

Anti-proliferative activity

Kaliyaperumal et al.,2024 [6] ; Heendeniya et al., 2020[31]

Flowers

Aqueous extract

In-vitro

Hypoglycemic and Hypolipidemic activity

Kaliyaperumal et al.,2024 [6]; Rangika et al.,2015[49]

Flowers

Aqueous extract

In-vitro

Hepato-protective activity

Kaliyaperumal et al.,2024 [6]; Wagh et al.,2010[85]

Flowers

Petroleum ether, Chloroform, and Ethyl acetate extracts

In vitro

Hepato-protective activity

Parekh et al.,2020[30] ;

Khatune et al., 2001[114]

Flowers

Chloroform extract

In-vitro

Larvicidal activity

Kaliyaperumal et al.,2024 [6]; Sah et al.,2012[75]

Flowers

Ethanolic extracts, Rengyolone 1 and its acetate derivative

In-vitro

Antifilarial activity

Parekh et al.,2020[30]   ;

Tuntiwachwuttikul et al., 2003[110]

Flowers

Zinc oxide nanoparticles synthesized using aqueous extract

In-vitro

Anti-fungal activity

Parekh et al.,2020[30]   ;

Jamdagni et al.,2018[37]

Flowers

Aqueous extracts

In-vivo

Hypoglycemic and hypolipidemic activity

Kaliyaperumal et al.,2024 [6] ; Parekh et al.,2020[30]   ; Rangika et al.,2015[49]

Stem Bark

Ethanol extract

In-vivo

Anti-diabetic activity

Kaliyaperumal et al.,2024 [6]; Suresh et al.,2010[86]

Stem Bark

Methanolic extract

In-vivo

Anti-inflammatory activity and Analgesic activity

Parekh et al.,2020[30] ;

Kakoti et al.,2013[60]

Stem Bark

Ethyl acetate extract

In-vivo

Anti-arthritic activity

Parekh et al.,2020[30] ;

Puri et al.,1994[125]

Stem Bark

Petroleum ether, Chloroform, and Ethanol extracts

In-vivo

Hepato-protective activity

Parekh et al.,2020[30] ;

Manisha et al.,2009[91]

Stem Bark

Ethanolic extract

In- vivo

Antibacterial activity

Parekh et al.,2020[30] ;

Suresh et al.,2010[86]

Stem Bark

Aqueous/Methanolic extract

 

Antioxidant activity

Parekh et al.,2020[30] ;

Thakur et al.,2017[40]

Seed-kernel

Iridoid glucosides

In-vitro

Antimalarial activity

Parekh et al.,2020 [30] ;

Shukla et al.,2012 [76]

Seeds

n-Butanol fraction of 50% ethanolic extract, Arbortristoside-A, and Arbortristoside C

In-vivo

Antiviral activity

Kaliyaperumal et al.,2024 [6]; Parekh et al.,2020 [30] ;

Gupta et al.,2005[108]

Seeds

Arbortristoside-A (AT) and 7-O-trans-cinnamoyl-6-hydroxyloganin (6-HL) ethanolic extract

In-vitro

& In-vivo

Antiulcer activity

Kaliyaperumal et al.,2024 [6] ; Mishra et al.,2013[59]

Seeds

Methanolic extract

In-vivo

Immunostimulatory activity

Kirubakaran et al.,2016 [6]; Parekh et al.,2020 [30]

Seeds

Chloroform extract

In-vivo

Immuno-modulator/ Immunorestorative activity

Kirubakaran et al.,2010 [6] ;

Parekh et al.,2020[30]

Seeds

Arbortristiside-A and 7-O-trans-cinnamoyl6?-hydroxyloganin

In-vivo

Anti-ulcerogenic activity/ Ulcer healing property

Parekh et al.,2020[30] ;

Mishra et al.,2013[59]

Fruits

Water-soluble fraction of 50% ethanolic extract

In-vivo

Antistress activity, Anti-oxidant activity, Antibacterial activity

Parekh et al.,2020[30] ;Tripathi et al.,2013[67]

Fruits

Methanolic extract

In-vitro

Anti-oxidant activity

Parekh et al.,2020[30] ; Kumari et al.,2012b[73]

Fruits

Petroleum ether and Methanolic extracts

In-vitro

Antibacterial activity

Parekh et al.,2020[30] ;

Shinde et al.,2014[55]

Leaves and Fruits

99% ethanolic extract

In-vitro

Antifilarial activity

Parekh et al.,2020[30] ;

Simonsen et al.,2001[112]

Fruits, Seeds, and Leaves

Water-soluble ethanolic extract

In-vivo

Immuno-modulator/ Immunorestorative activity

Parekh et al.,2020[30] ;

Rathore et al.,2007 [100]

Leaves and Stem Bark

Hydro-alcoholic extract

In-vitro

Antibacterial activity

Parekh et al.,2020[30] ;

Satyal et al.,2011[80]

Leaves, Flower, Fruits, and Seeds

Ethyl acetate and Chloroform extract

In-vivo

Hepato-protective activity and Antibacterial activity

Parekh et al.,2020[30];

Priya et al.,2007[103]

Flowers, leaves and Seeds

50% ethanolic extract of seeds, flowers and leaves, aqueous fractions

In-vivo

Immuno-stimulant activity

Rajput et al.,2024[3] ;

Puri et al.,1994[125]

Leaves, Seed , Flower, Stem, and Root

50% ethanolic extract

In-vivo

Antipyretic

Parekh et al.,2020[30] ;

Khan et al.,1995[123]

Whole plant

80% methanolic extract

In-vivo

Immunostimulatory activity

Parekh et al.,2020[30];

Bhalerao et al., 2011[78]

Whole plant material

Aqueous, Ethanol, Benzene, Petroleum ether, and Chloroform extracts

In vitro

Antibacterial activity

Parekh et al.,2020[30];

Aggarwal et al.,2013[66]

Root barks

Aqueous, Ethanolic, Petroleum ether, and Chloroform extracts

In vitro

Hepato-protective activity

Parekh et al.,2020[30] ;

Verma et al.,2011[78]

Table -3, Anticancer and Antitumor, Cytotoxic activity of flowers aqueous extract of NAT:

 

Investigated

Plant parts

Used in Studies Extracts

Study plan

Pharmacological

Activities

References

Leaves

Ethanolic extract

In-vivo

Tumour necrosis factor Depleting activity

Parekh et al.,2020[30] ;

Paul et al.,1997[120]

Leaves

Methanolic extract

In-vitro, A-549 cancer cell line

Anticancer activities

Parekh et al.,2020[30] ;

Kumari et al.,2017[39]

 

Flowers

Crude aqueous extracts- Chloroform  fraction,

Ethyl acetate fraction,

Hexane fraction

In-vitro

Anticancer activities

Kaliyaperumal et al.,2024 [6] ; Heendeniya et al., 2020[31]

Flowers

Ethyl acetate, Ethanolic and aqueous extracts

In-vitro

Anticancer activities

Parekh et al.,2020[30] ;

Khanapur et al.,2014[57]

Dried fruit

Methanol extract

In-vivo

Anticancer activities

Kaliyaperumal et al., 2024 [6] ; Gulshan et al.,2015[50]

Fruits, leaves, and Stem barks

Methanol extract

In-vivo

Anticancer (human breast cancer) activities

Kushwah et al.,2023[18] ;

Khatu et al.,2001[114]

Flowers

Flower extracts synthesized ZnO nanoparticle

In vitro

Anticancer activities

Swain et al.,2024[4] ;

Chabattula et al.,2024[5]

Flowers

Petroleum ether, Chloroform, and Ethyl acetate extracts

In vitro

Cytotoxic activities

Kaliyaperumal et al., 2024 [6]; Parekh et al.,2020[30];

Khatune et al.,2001[115]

Flowers

Ethyl acetate and ethanol extracts

In-vivo

Antiproliferative, Anticancer activities

Swain et al.,2024[4] ;

Khanapur et al.,2014[57] ; Susan et al.,1986[132]

Flowers

Crude aqueous extracts

In vitro

Cytotoxic activities

Naznin et al.,2001[111]

Leaves and Stem

Successive extraction using hexane and ethanol

In-vitro

Cytotoxic activities

Parekh et al.,2020[30] ;

Chidi et al.,2015[51]

Stem, leaves, and Fruits

Methanolic extract

In-vitro

Cytotoxic activities

Parekh et al.,2020[30] ;

Kumari et al.,2012b[73]

 

Table - 4: Analysis of Microbial load (By WHO/AOAC/AYUSH/API/UPI Std. Methods)

 

S. N0.

Parameter Analyzed

 

Results

 

WHO Limit

NAT - 1

NAT - 2

NAT - 3

1

Total Bacterial Count

580 cfu/gm

584 cfu/gm

584 cfu/gm

105cfu/gm

2

Total Fungal Count

630 cfu/gm

636 cfu/gm

632 cfu/gm

103cfu/gm

3

Escherichia coli

Absent

Absent

Absent

Absent

4

Salmonella typhai Spp.

Absent

Absent

Absent

Absent

5

Staphylococcus aurous

Absent

Absent

Absent

Absent

Table - 5: Estimation of Heavy Metals ( By AAS-GF)

 

S. N0.

Parameter Analyzed

 

Results

 

WHO Limit

NAT - 1

NAT - 2

NAT - 3

1

Lead

3.02ppm

3.03ppm

3.04ppm

10ppm

2

Cadmium

0.04ppb

0.03ppb

0.04ppb

0.3ppm

3

Mercury

N/D

N/D

N/D

1.0ppm

4

Arsenic

0.05 ppm

0.05 ppm

0.04 ppm

3.0ppm

 

Table - 6: Estimation of Aflatoxins (By HPTLC)

 

S. N0.

Parameter Analyzed

 

Results

 

WHO Limit

NAT - 1

NAT - 2

NAT - 3

1

Aflatoxin, B1

N/D

N/D

N/D

0.5ppm

2

Aflatoxin, B2

N/D

N/D

N/D

0.1ppm

3

Aflatoxin, G1

N/D

N/D

N/D

0.5ppm

4

Aflatoxin, G2

N/D

N/D

N/D

0.1ppm

 

Table - 7, Estimation of Pesticide Residues (By GC-MS):

 

S. N0.

Parameter Analyzed

 

Results

 

WHO Limit in ppm.

NAT - 1

NAT - 2

NAT - 3

1

DDT (all isomers, sum of ?, ?’-DDT, ?, ?’ DDT, ?, ?’-DDE and ?, ?’-TDE (DDD expressed as DDT)

N/D

N/D

N/D

1.0

2

HCH (sum of all isomers)

N/D

N/D

N/D

0.3

3

Endosulphan (all isomers)

N/D

N/D

N/D

3.0

4

Azinphos-methyl

N/D

N/D

N/D

1.0

5

Alachlor

N/D

N/D

N/D

0.02

6

Aldrin (Aldrin and dieldrin combined expressed as dieldrin)

N/D

N/D

N/D

0.05

7

Chlordane (cis& tans)

N/D

N/D

N/D

0.05

8

Chlorfenvinphos

N/D

N/D

N/D

0.5

9

Heptachlor (sum of heptachlor and heptachlor epoxide expressed as heptachlor)

N/D

N/D

N/D

0.05

10

Endrin

N/D

N/D

N/D

0.05

11

Ethion

N/D

N/D

N/D

2.0

12

Chlorpyrifos

N/D

N/D

N/D

0.2

13

Chlorpyrifos-methyl

N/D

N/D

N/D

0.1

14

Parathion methyl

N/D

N/D

N/D

0.2

15

Malathion

N/D

N/D

N/D

1.0

16

Parathion

N/D

N/D

N/D

0.5

17

Diazinon

N/D

N/D

N/D

0.5

18

Dichlorvos

N/D

N/D

N/D

1.0

19

Methidathion

N/D

N/D

N/D

0.2

20

Phosalone

N/D

N/D

N/D

0.1

21

Fenvalerate

N/D

N/D

N/D

1.5

22

Cypermethrin (including other mixtures of constituent isomers sum of isomers)

N/D

N/D

N/D

1.0

23

Fenitrothion

N/D

N/D

N/D

0.5

24

Deltamethrin

N/D

N/D

N/D

0.5

25

Permethrin (sum of isomers)

N/D

N/D

N/D

1.0

26

Pirimiphos methyl

N/D

N/D

N/D

4,0

N/D Where:=Not Detect

CONCLUSIONS:

The investigated and revalidated 3 samples of Arial plant part of NAT. drug were found to be of very good quality and devoid of any impurities or free from any hazardous, toxic contamination and adulterations according to the drug Authentications, identifications quality control revealed results data’s basis. The ranges of all the Botanical, Pharmagnistical, Biodiversity, Toxicological by Satandared methods of WHO/AOAC/AYUSH/API/ UPI,  and AAS-GF, HPTLC,GC-MS investigated profiling research data’s constants used for the quality analysis of the entire NAT arial part 3 samples of the plant are normal. Numerous secondary metabolites have been detected and reconfirmed in the Botanical, Pharmagnistical, Biodiversity, Toxicological research by investigation, Analyzed and applied advance methods and sophisticated instruments techniques. The Toxicology investigation potential research data’s have been shown that thus the arial part’s of the NAT feet for internal use as a drug supplement. The herbaceous medicinal plant NAT has been climatically very rich form in wild, worldwide biodiversity occurrence. As a result of its potent quality, safety and toxicity studies of properties, NAT may be treat and therapeutically used in various ASU traditional and alternative medicines preparations since ancient time. Investigated In-vitro ,In-vivo research reports data’s confirmed therapeutic medicinal potent as a Anticancer activities, Cytotoxic activities, Antiarthritis, Antistress, Antimicrobial, Antifungal, Antibacterial, Antioxidant, Anti-Inflammatory, Hypoglycemic and hypolipidemic activity, Antiviral activity, Antiulcer activity, Analgesic activity, Obstinate Sciatica disorder’s from since ancient time. Investigated NAT - ASU drug can be incorporated and support to developed revalidation of novel drug development, investigation of novel bioactive marker constituents, compounds mechanism of actions, pharmacopoeial standard and atlas monographs, Pharmacovigilance aspects of NAT. However, Further studies are required to identify, isolate, and elucidate the structures of novel bio-active constituents present in NAT apply upon GC-MS, LC-MS, XRD, SEM-EDX advance sophisticated instruments techniques of these investigated drug scan still be carried out for purposes of advance research. that contribute to its health benefits. Besides, more evidence, especially through In-vivo and clinical trials, is necessary to determine the mechanisms involved in the bioactivities mentioned previously. Even though there are available preliminary data on the medicinal and nutritional values of NAT stem bark, root bark, flower, seeds parts information is still lacking, especially on other parts of the plant, such as the arial leaves, stems bark. thus, studies on the these plant parts of NAT also be done to discover potential bioactive compounds that can be exploited for discover novel drug development and health advantages.

Limitations and Future Remarks of the Study:

The present studies data’s of  Botanical, Pharmagnistical, Biodiversity and Toxicological profiles shown the reconfirmation and presence of QC, QA, DSR, PV of arial part samples of NAT. In the future, investigated data may be used to Advance revalidation pharmacopoeial Drug Standardization Research, Pharmacopeial atlas monographs profiling development, novel drug development, investigation of novel bioactive marker constituents, compounds mechanism of actions, Pharmacovigilance aspects and confirm these investigated resulted data’s.

Ethical approval:

As the work is purely an In-vitro study, ethical clearance is not required.

Author’s contributions:

Dr Pawan Kumar Sagar (Chemistry): Manuscript work designed, Carried out Instrumental, Chemistry part and Manuscript written and revised manuscript review. Dr. S. Sajwan, Dr. K.Venkatesan, Mrs. Jayanthy A, Dr. Mukesh Kumar and Dr. R M Singh: provide very helpful Support and Carried out Botanical, Pharmacognosy, taxonomy and biodiversity climatic occurrence, confirmation and authentication works designed and Research Material’s Collection. Devi P M Sri, and Mr. S. Kashyap (Chemistry): carried out Toxicology, Microbiological and Analytical data analysis.

Declaration of Competing Interest:

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

ACKNOWLEDGEMENTS:

The authors are grateful thanks to higher related senior authority Director General of CCRUM, Ministry of AYUSH, New Delhi, Scientific Researcher supporting staff’s of the Central Council for Research in Unani Medicine, Ministry of AYUSH, Government of India, and Regional Research Institute of Unani Medicine, Chennai (NABH and NABL Accredited, certified) and Drug Standardization Research Institute, PCIM&H Campus, IInd floor, Kamla Nehru Nagar, Ghaziabad UP.( NABL ISO-17025-2017,Accredited, certified), PCIM&H, Ghaziabad UP. India for provided all relevant cooperation, supports of required modern and scientific facilities to conduct this research work.

Financial support statement:    Nil.

Main Author ORCID ID:  Dr. Pawan Kumar Sagar : 0009-0007-8695-9958

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Dr.Pawan Kumar Sagar
Corresponding author

Drug Standardisation Research Institute, (CCRUM, M/o AYUSH, Govt. of India), PCIMU&H Campus, Kamla Nehru Nagar, Ghaziabad- 201002, UP. INDIA

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Sajwan S.
Co-author

Drug Standardization Research Institute, (Under CCRUM, Ministry of AYUSH., Govt. of India), PCIM&H Campus, IInd Floor, Kamla Nehru Nagar, Ghaziabad, U.P., India

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Sri P. M. D.
Co-author

Regional Research Institute of Unani Medicine, (Under CCRUM, Ministry of AYUSH) Govt. of India, Royapuram, Chennai, TN, India

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Kashyap S.
Co-author

Drug Standardization Research Institute, (Under CCRUM, Ministry of AYUSH., Govt. of India), PCIM&H Campus, IInd Floor, Kamla Nehru Nagar, Ghaziabad, U.P., India

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Venkatesan K.
Co-author

Regional Research Institute of Unani Medicine, (Under CCRUM, Ministry of AYUSH) Govt. of India, Royapuram, Chennai, TN, India

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Jayanthy A.
Co-author

Pharmacopoeia Commission for Indian Medicine & Homoeopathy (PCIM&H), Kamla Nehru Nagar, Ghaziabad UP. India (Ministry of AYUSH, Govt. of India)

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Kumar M.
Co-author

Pharmacopoeia Commission for Indian Medicine & Homoeopathy (PCIM&H), Kamla Nehru Nagar, Ghaziabad UP. India (Ministry of AYUSH, Govt. of India)

Pawan Kumar Sagar*, Sajwan S., Sri P. M. D., Kashyap S., Venkatesan K., Jayanthy A., Kumar M., Scientific DSR Validation, Pharmacognostical, Biodiversity, Toxicological Research Studies of Nyctanthes Arbortristis L.- Aerial Leaves Part and Their Pharmacological, Therapeutic Medicinal Values, Int. J. of Pharm. Sci., 2025, Vol 3, Issue 1, 1641-1669. https://doi.org/10.5281/zenodo.14695813

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