INTRODUCTION:

In recent years, the usage of herbal plants and their parts gained major attention among natural product researchers as a vital source of medicinal agents and cosmetics due to easy accessibility, friendly, economic, and no or fewer side effects.

According to World Health Organization 2014, it was stated that in most developing nations, plants-based drug plays an essential role in meeting the major healthcare needs of the people and highlights specific types of its treatment¹. According to Chanda et al., adulteration occurs when an original plant is replaced or added with another plant material as well as any foreign substance for the purpose of increasing the potency of the product or reducing its cost². Standardization is an initial stage and essential to ensure a predefined amount of quantity, quality and therapeutic effect of ingredients in each dose³. To establish the standardization and authentication parameters of crude drugs of natural origin, pharmacognostic studies of the medicinal plant are considered vital. In such consideration, Christia obcordata (C.obcordata) and Christia vespertilionis (C. vespertilionis) have gained great attention among researchers as these plants show to have potential health benefits. Christia is a legume belongs to the Fabaceae family comprising of 13 varieties that is found in Malaysia, Australia and tropical Southeast Asia. In these countries it is initially observed as an ornamental plant⁴. Every species of Christia has unique butterfly-shaped and heart shaped leaves. Christia obcordata (Poir.) Bakh.f. (syn. Ploca humilis, Hedysarum obcordatum, Desmodium obcordatum, or Lourea obcordata) is also known as green butterfly plant with kidney- shaped and flutter like butterfly wings and can grow up to 1 metre. The plant has trifoliate leaves and slender stems. The juvenile background leaves were light green and became bright green with dark burgundy stripes when matured⁵. Christia vespertilionis (L. f.) Bakh. F., (syn. Hedysarum vespertilionis, Lourea vespertilionis), is known as red butterfly wing, mariposa (butterfly in Spanish) or “rerama” (butterfly in Malaysia)⁶. It can grow up to 1.2 metre in height and has trifoliate leaves and slender stems. The leaves of the plant are purple in colour and dark green colour with pale green stripes along prominent veins when the leaf is matured. Traditionally C. vespertilionis has been used to treat cold muscle weakness, poor blood circulation, bronchitis, and inflamed tonsils. C.obcordata is also reported to be used to treat acute, chronic nephritis and urinary blockage⁵. The picture of the plants was shown in figure 1.

Figure 1: Christia obcordate (left) and Christia vespertilionis (right)

In a study, it was reported that these plants are rich in triterpenes, alkaloids, fatty acids, phenols, alkanes⁷. A Study report from Smitha et al., showed the phytochemical analysis of methanolic leaves of C. vespertilionis confirms the presence of alkaloids, flavonoids, glycoside, tannin, diterpenes, coumarin, and quinine⁸. Furthermore, a study conducted by Harish et al., reported thirteen compounds were identified in this plant which are two of flavonoid glycosides (quercetin-3-O-glucoside and catechin-3-O-β-D glucopyranoside), three of isoflavonoids (2’-hydroxy genestin, orobol and 2,3-dihydro-2’-hydroxy-genestin), and three of pentacyclic triterpenes (D:C- friedoolean-8-en-29α-ol), ursolic acid methyl ester, oleanolic acid methyl ester, and erythrodiol). In addition, three steroids (stigmasterol, β-sitosterol acetate and β-sitosterol) and a monoterpene called geraniol were identified. Furthermore, 7-isopropylidene-1-methyl-1,2,6,7,8,9- hexahydronapthalene (called as christene) and 2- hydroxydecanylpentadec-5, 8,10, 12- tetraenoate (called as christanoate) were also reported in the plant⁹. The compound christene has shown significantly active (IC₅₀= 9.0µg/ml) against Plasmodium falciparum⁴.

In Malaysia, C. vespertilionis leaves decoction was believed to treat cancer. Furthermore, the study reported that rubbing the leaves of this plant on the affected area helps in healing the scabies¹⁰. The Whole plant was believed to treat tuberculosis and snake bite while the leaves of this plant can be used as a topical treatment to heal bone fractures¹¹. The plant is also reported for its potency in increasing blood circulation, healing muscle weakness, treat bronchitis and colds and the study on, different parts of C. vespertilionis decoction (methanol and aqueous-methanol) of the roots, leaves and stems have been evaluated in vitro for its antiplasmodial potential against Plasmodium falciparum NF-54¹². Based on the reported benefits this herbal plant was in high focus among the researchers. However, currently there is no published informations regarding the standardization and authentication of C.obcordata and C. vespertilionis. Thus, this study will focus on pharmacognostic characteristics of the selected plants and will increase its visibility to study more on their effectiveness in the field of herbal medicine. Moreover, there is less degree research has been done regarding standard parameters for these plants.

MATERIALS AND METHODS:

Collection and preparation of plant material:

The fresh plant materials of C.obcordata was collected from Hock Loke Siew Nursery, Ipoh, Perak and C. vespertilionis was collected from Asow Crafts and Landscaping, Ipoh Perak. The specimen was authenticated by Madam Fadhilah binti Abdul Rani, Deputy director of Kompleks Pertanian Telok Chengai, Kedah. Upon confirmation, the aerial parts of the plants were cleaned, shade dried, and grounded using a mechanical blender. The powdered materials of the plants were sieved seperately and stored individually in an airtight container until further use.

Macroscopic evaluation:

Macroscopic studies like shape, colour, size, texture, margin, odour, taste, and phyllo taxis were studied using fresh plant materials like leaves and stems¹.

Microscopy evaluation:

a) Transverse section of plant leaves:

The thinnest section of fresh leaves and stems of both plants were separately stained with a mixture of 1:1 phloroglucinol and concentrated hydrochloric acid. After that, the stained samples were placed onto the slide followed by a few drops of glycerin added to give a moisture effect and covered with coverslip respectively. The slide was then observed under a binocular optical microscope, a clear picture was captured using (Leica DM750, Germany) microscope fitted with a digital camera¹³.

b) Powder study:

Dried leaves powder was taken instead of a fresh section of the plant similar to microscopic study. Before staining, the powders were heated in a mixture of 5g chloral hydrate and 100ml of distilled water. Staining agents such as concentrated hydrochloric acid, phloroglucinol N/20 iodine solution at the ratio of 1:1 was used to study the powder. Distilled water was used to identify the presence of cellular components.

c) Determination of Plant constant:

The midrib of the leaf was removed by boiling with a 10% chloral hydrate solution. Using the forceps, the upper and lower epidermis were peeled out separately, kept on the slide, and mounted in glycerine water. To determine the plant constant following tests like palisade ratio, vein termination number and vein islet number, and stomatal index were studied¹⁴.Stomatal index present in 1 sq. were counted using the following formula:

S. I = S/E + S x 100

Where

S. I = Stomatal Index,

S = No. of stomata per unit area,

E = No. of epidermal cells in the same unit area

d) Palisade ratio: It is the mean number of palisade cells underneath each epidermal cell. The test results will give subjective information to distinguish the characteristics of a closely related species¹⁵.

e) Vein termination number and vein islet number: Veinlet termination number was calculated using average number of vein-islets per square mm of a leaf surface. It was determined by counting the number of vein-islets in an area of 4 square mm of the central part of the leaf between the midrib and the margin¹⁶.

f) Extraction: About 2g of each dried plant material was extracted using petroleum ether, chloroform, methanol, and water (increasing in the order of polarity) in an ultrasonicator for 30 minutes. Obtained extracts were used to perform preliminary phytochemical screening to identify the phyto constituent of the plant material¹⁷.

Qualitative Preliminary Phytochemical Screening: Preliminary phytochemical screening was performed on various extracts on both plants and the aerial parts. The secondary metabolites like alkaloid, carbohydrates, proteins, tannins, steroid saponins, and flavonoids were tested^18,19.

Fluorescence Analysis:

The fluorescence characteristics of all extracts were studied under ultraviolet light at short wavelength (254nm) and long wavelength (365nm)^20,21.

RESULTS AND DISCUSSION:

Macroscopic evaluation:

The macroscopic features of the leaves of C.obcordata and C.Vespertilionis were examined and recorded as shown in figure 2.

Figure 2: Macroscopic characteristics of C. obcordata and C. vespertilionis

Macroscopic evaluation is a method that is used to study the morphological characteristics of the plant part which can be observed by our naked eye or through a magnifying lens. Macroscopic evaluation is considered an important aspect of herbal analysis for the correct identification of crude drugs. Based on the observation made, both leaves show similar characteristics in the followings, shape-obcordate, phyllo taxis – alternate, margin-entire, leaf type- compound, leaf veination – recticulate, odour- no fragrance. However, colour of the leaf for C.obcordata was found to be green and permanently veined with deep maroon stripes on the other hand C.vespertlionis showed purple red tones, prominently veined with shades of pink.

Microscopic evaluation:

Transverse section of the leaf:

A Transverse section on the leaves of C. obcordata and C. vespertilionis was examined and shown in Figure 3 and 3.1. The transverse section of the C.obcordata leaf shows single layered barrel shaped epidermal cells, arranged compactly with numerous unicellular covering trichomes and a thin cuticle. The leaf structure demonstrates the dorsiventral nature of the leaves that represents the presence of palisade cells below the upper epidermis only. Crystal sheaths and starch grains are dispersed in the collenchyma and parenchyma cells. Vascular bindle is leptocentric where the layer of xylem vessels surrounds the phloem fibres. The transverse section of the leaf shows single layered barrel shaped epidermal cells on both surfaces covered with thin cuticles. The cells are arranged closely without any intercellular spaces in between them. These cells are devoid of chloroplasts. The presence of uniseriate multicellular covering trichomes were seen. Paracytic stomata are seen on the epidermis. The mesophyll is a ground green tissue present in between upper epidermis and lower epidermis. The leaf structure demonstrates dorsiventral nature that represents presence of the palisade cells below the upper epidermis only. Palisade cells are elongated, arranged in 3-4 layers, compact, and contain greener chloroplasts. Crystal sheaths and starch grains are dispersed in the collenchyma and parenchyma cells. The mid rib portion consists of a vascular bundle, collenchyma, and parenchyma. There are discontinuous palisade cells in the mi rib region. Vascular bindle is leptocentric where the layer of xylem vessels surrounds the phloem fibers. Xylem vessels are arranged in groups and a layer of collenchyma is seen below the layer of xylem vessels. A layer of collenchymatous cells is seen below the vascular bundle.

Figure 3: Transverse section of C.obcordata leaf.

a- Collenchyma, b- Upper cuticle, c- Upper epidermis, d- Palisade cell, e- Xylem vessel, f- Parenchyma, g- Phloem fibre, h- Pericyclic fibre, i- Lower epidermis, j- Lower cuticle, k- Phloem fibre, l- Collenchyma, m- covering trichome.

The transverse section of C. vespertilionis leaf consists of an upper and lower epidermis with single layered closely arranged almost isodiametric and rectangular cells with no inter cellular spaces in between them. Chloroplasts cannot be seen in these cells. Both upper epidermis on their outer surface is covered by a thin cuticle. The lower epidermis appeared wavy in outline and after epidermis, chlolenchyma cells were present.

Figure 3.1. Transverse section of C.vespertilionis leaf.

a- Upper cuticle, b- Upper epidermis,c-Trichomes, d- Palisade cell, e- Spongy mesophyll, f- Parenchyma, g- Collenchyma, h- Sclerenchyma, i- Lower epidermis, j- Lower cuticle, k- Xylem vessels, l-Phloem.

Paracytic (Rubiaceous) stomata have two subsidiary cells, of which the long axes parallel to the axis of the stoma and are seen on both surfaces. However, the distribution of stomata on the epidermal surface is hypostomatic indicating the presence of a greater number of stomata on the lower surface as compared to the upper surface. Numerous uniseriate multicellular covering trichomes were seen on both epidermal layers. The mesophyll is a ground tissue present between the upper epidermis and lower epidermis. The mesophyll tissue consists of palisade cells and spongy parenchyma which are well differentiated. The palisade cells are present only below the upper epidermis and reveal the dorsiventral nature of the leaf. The palisade parenchyma is discontinued at the midrib region and shows 2-3 layers of palisade cells closely packed without any intercellular space between them. The palisade cells are filled with many chloroplasts. The spongy parenchyma region consists of 4-5 layered parenchyma cells which are spherical, and isodiametric with the presence of inter cellular space between them.The midrib consists of a vascular bundle, collenchyma and sclerencyma tissue. The vascular bundle consists of xylem vessels and phloem fibers and is embedded at the center of the mid rib. Xylem tissue of the vascular bundle is present towards the upper epidermis and the phloem tissue towards the lower epidermal surface of the leaf. Bundle sheath is absent. A few patches of collenchyma cells are present towards both the dorsal and ventral surfaces close to the upper and lower epidermal ends.

Transverse section of stem:

The transverse section of C.obcordata stem shows single layered barrel shaped epidermal cells, arranged compactly with numerous unicellular covering trichomes and a thick cuticle. The vascular bundle has lignified vessels with outer phloem. The hypodermis is composed of 3 to 4 layers of collenchymatous cells. The endodermis is distinct and embedded composed of collenchymatous tissue with numerous pericyclic fibers arranged in groups at regular intervals. The vascular bundle is hadrocentric (amphicribral) where the xylem vessels are surrounded by phloem fibres. Phloem is broad and consists of phloem fibers in groups, sieve tubes, and companion cells. Xylem consists of vessels, tracheids, fibers, and xylem parenchyma. Medullary rays are 3 to 4 cells broad and radiating. The central portion is occupied by pith consisting of isodiametric parenchymous cells and contain starch granules. Figure 4 and 4.1 show the transverse section of the stem, and Iodine stain for starch identification respectively.

Figure 4: Transverse section of C.obcordata stem where, a:Medullary rays, b: Phloem, c: Xylem, d: Covering trichome, e: Epidermis, f: Cuticle, g: Pericyclic fibre, h: Pith, i: Collenchyme.

Figure 4. 1. Iodine stained transverse section of C.obcordata stem showing the presence of Starch.

The transverse section of C.vespertilionis stem reveals the presence of epidermis with a single layer of cells coated with a thin and striated layer of cuticle. Numerous uniseriate covering trichomes emerge from the epidermal layer and are found to be unicellular. The cortical region is formed by a few layers of collenchyma followed by 4 – 5 layers of sclerenchyma cells. Below the layer of sclerenchyma cells, bundles of phloem fibers are arranged with a few layers of sclerenchyma cells. Starch grains are seen in the endodermis. The vascular bundle is conjoint, consisting of the xylem and phloem that lie opposite to each other on the same radius. Isolated sclerenchyma fibers are located adjacent to the phloem differentiating multiple vascular bundles from each other. The medullar region consists of parenchyma and inclusions are found in some. The outer part consists of a single layer of epidermal cells made of parenchyma cells and covered with a thick cuticle. The cells are barrel shaped and compactly arranged with no intercellular spaces and chloroplasts. Several uniseriate covering trichomes emerge from the surface of the epidermis. The cortex is present below the epidermis in several layers and can be differentiated into the hypodermis and general cortex. The hypodermis is made up of a few layers of collenhyma cells. Underneath the epidermis, there is a ring of sclerenchymatous cells which surrounds the central structure. Cortex has collenchymatous hypodermis regularly or as discontinuous patches. The pericycle is composed of sclerenchyma and intervening masses of parenchyma as irregular patches (heterogeneous). The vascular bundles are wedge shaped, collateral, and arranged in a ring surrounding the pith. Cambium is absent. Each bundle is composed of an outer phloem and inner xylem on the same radius. Pith is extensively developed and occupies the central portion of the ground tissue. It is made from rounded or oval, thin-walled parenchymatous cells with distinct intercellular spaces. Figure 5 represents the transverse section of C.vespertilionis stem.

Figure 5. Transverse section of C.vespertilionis stem where, a:Xylem vessels, b: Sclerenchyma, c: Pholem, d: Parenchyma, e: Endodermis, f: Epidermis, g: Epidermis with cuticle, h: Collenchyma, i: Covering trichomes.

Powder microscopy of the levaes of C.obcordata and C. vespertilionis:

Powder microscopy of both plant leaves was observed individually and recorded in Figure 6 and 7. Powder microscopy of C.obcordata and C.vespertilionis revealed the presence of uniseriate multicellular covering trichomes, calcium oxalate crystals, xylem vessels, phloem fibres. However, in addition powder microscopy of C.vespertilionis leaves showed the presence of stomata, covering trichomes with epidermal cells, parenchyma cells, and sieve tubes.

Figure 6: Powder microscopy of C.obcordata leaves

Figure 7: Powder microscopy of C.vespertilionis leaves

Determination of plant constant:

The plant constant was evaluated on both the plant leaves for their stomatal number and stomatal index, vein islet number, vein termination number, and palisade ratio were also observed. From the results it was found that C.vespertilionis have higher plant constant values in most studied parameters like Stomatal number (Upper epidermis)-9.1-9.8-10.6,Stomatalnumber (Lower epidermis) -19.8-21.4 - 24.3,Stomatalindex(Upper epidermis)-12.5-14.9-16.6,Stomatal index (Lower epidermis)- 25.6-27.4-30.7, Vein islet number-5.7-7.2-10.2, Vein termination number-8.6-9.4-10.3, Palisade ratio-12. In contrast, C.obcordata has showed higher values on the stomatal number and stomatal index at their lower epidermis. Following are the plant constant values of C.obcordata, Stomatal number (Upper epidermis)-4.4-5.8-7.2,Stomatal number (Lower epidermis)-28.6-29.8-31.7,Stomatal index (Upper epidermis)-5.2-6.1-6.8, Stomatal index (Lower epidermis)- 28.6-29.8-31.7, Vein islet number-4.8-5.3-8.6, Vein termination number-7.8-8.3-9.5, Palisade ratio-8. The bolt number is the average of the plant constant values.

Preliminary Phytochemical screening:

The aerial parts of the plant were extracted and tested for their phyto constituents, and the results were presented in Table 1. The plant aqueous extract confirmed for the presence of carbohydrates and proteins, petroleum ether and chloroform extracts confirms presence of steroids, and in methanol aqueous extract tannin, saponin, and flavonoids were present. However, proteins were found to be absent in both plants.

Physiochemical parameters:

Ash values and extractive values:

The aerial parts of C.obcordata and C.vespertilionis were evaluated for their ash value and extractive values on the dry basis of the sample, total ash value, acid insoluble ash, and water-soluble ash values were observed to be 5.1% w/w, 4.9% w/w, and 3.9% w/w for C. vespertilionis however extractive values of water and ethanol was found to be 15% w/w and 11% w/w. In contrast with similar conditions, C. obcordata showed total ash value of 5.4% w/w, acid insoluble ash value of 5.3% w/w, and water-soluble ash value of 4.1% w/w, extractive values of water and ethanol were found to be 17% and 13% w/w respectively.

Fluorescence Analysis of the Plant Extracts:

To confirm the doubtful specimen, fluorescence analysis on the powdered plant sample can be used. In the current study, aerial parts of both plants were extracted with 4 solvents and evaluated in three conditions such as short ultra-violet light (254nm), long ultra-violet light (365 nm), and visible light which has shown different colors. The results of fluorescence analysis were revealed in Table 2 and the image in Figure 8 (A-F).

Observation of different chemical constituents in the crude plants can be analysed by using fluorescence analysis. Specific constituents in the plant exhibit fluorescence characteristics when tested with UV light or with the visible light. The changing in colour from visible light to ultraviolet light can be served as a marker for the existence or lack of chemical constituents as organic molecules absorb light over a specific range of wavelengths and reemit radiations. Hence, fluorescence analysis turns into one of the vital considerations in determining the purity and quality of crude drugs.

Table 1: Preliminary phytochemical screening on the aerial parts extract of C.obcordata and C.vespertilionis.

Chemical constituents	Chemical tests	P.E.E C.o	C.E C.o	M.E C.o	D.W.E C.o	P.E.E C.v	C.E C.v	M.E C.v	D.W.E C.v
Steroid	Libermann Burchard test	+	+	-	-	+	+	-	-
Flavanoid	Shinoda test	-	-	+	+	-	-	+	+
Tannin	Ferric Chloride test	-	-	+	+	-	-	+	+
Alkaloid	Wagner’s test	-	-	+	-	-	-	+	-
Saponin	Foam test	-	-	-	-	-	-	-	-
Carbohydrate	Molisch test	-	-	-	+	-	-	-	+
Protein	Biuret test	-	-	-	-	-	-	-	-

Where, P.E.E – Petroleum ether extract, C.E -Chloroform extract, M.E – Methanol extract, D.W.E -Distilled water extract, C.o - C.obcordata and C.v - C.vespertilionis. ‘+’ = Present ‘-‘=Absent.

Table 2: Results of fluorescence analysis on the powdered aerial parts of C.vespertilionis and C.obcordata.

Reagents

C.vespertilionis

C.obcordata

Color observed in day light

Color observed under short ultraviolet light (254 nm)

Color observed under long ultraviolet light (365 nm)

Color observed in day light

Color observed under short ultraviolet light (254 nm)

Color observed under long ultraviolet light (365 nm)

Petroleum ether

Chloroform

Methanol

Aqueous

Pale yellow

Colourless

Pale buff colour

Colourless

Orange (F)

Red (F)

Purple (F)

Colourless

Pale green

Pale yellow

Pale green

Deep green

Pale yellow

Light green (F)

Deep red (F)

Pale brown (F)

Colourless

F = Fluoresence.

CONCLUSION:

To conclude, the findings of current study on the aerial parts of C.vespertilionis and C.obcordata will be considered an important tool to provide valuable information about standardization of these plants. Furthermore, the current standardization is beneficial for identifying adulterant specimens and facilitates compiling of suitable monographs for appropriate documentation purposes in the field of herbal medicine. Detailed research of the morphological and microscopical characters will be used in determining and enhancing in differentiating plants of identical or various species.

CONFLICT OF INTEREST:

There is no conflict of interest.

ACKNOWLEDGEMENT:

The authors would like to thank the management and laboratory department of Faculty of Pharmacy and Health Sciences Royal College of Medicine Perak, Universiti Kuala Lumpur in providing necessary facilities and support for the completion of this research work.

REFERENCES:

1. Dash GK. Hashim MHB. Hassan AKR. Ravindran M. Pharmacognostic Studies on the Leaves of Annona muricata Linn. Pharmacog J. 2021; 13(1): 1-7. https://doi.org/10.5530/pj.2021.13.34

2. Chanda S. Importance of pharmacognostic study of medicinal plants: An overview. Jounal of Pharmacognosy and Phytochemistry. 2014; 2(5): 69-73.

3. Swapnil G. Patil AS. Standard Tools for Evaluation of Herbal Drugs: An Overview. The Pharma Innovation. 2013; 2(9): 60-61.

4. Dash GK. An Appraisal of Christia vespertilionis (L.F.) Bakh F.: A Promising Medicinal Plant. International of Journal of Pharmacognosy and Phtochemical Research. 2016; 8(6): 1037-1039.

5. Whiting PA. Commercial Production of Christia subcordata Moench by Establishing Clutural Practices and Applying Plant Growth. University of Georgia Theses and Dissertations. Retrieved from https://athenaeum.libs.uga.edu/handle/10724/24521. Accessed on 8th Nov 2022.

6. Ravindran M. Dash GK. Sengamalam R. Vignesh R. Red Butterfly wing (Christia vespertilionis (L. f) Bakh F.): a potential anticancer and antiplasmodial herb under threat of survival in Malaysia. Malays. J. Med. Health. Sci. 2019; 15(3): 154.

7. Daniela Hofer GSW. Christia vespertilionis plant extracts as novel antiproliferative agent against human neuroendocrine tumor cells. Spandidos Publications. 2013; 29(6): 2219-2226. https://doi.org/10.3892/or.2013.2367

8. Smitha SRJ. Anatomical Profiling and Phytochemical Analysis of Christia vespertilionis (L.F.) Bakh. F. International Journal of Pharmacy and Biological Sciences. 2019; 9(1): 40-50.

9. Harish C. Upadhyay BS. Novel Antiplasmodial Agents from Christia vespertilionis. Natural Product Communications. 2013; 8(11): 1591-1594. https://doi.org/10.1177/1934578x1300801123

10. Garnock-Jones P. Note on some folklore drugs and remedies of the Law group, FIJI. South Pac J Nat Sci.1983; 4: 4-8.

11. Retrieved from http://www.efloras.org. Flora of China 10: 289-290. 2010. CHRISTIA Moench, Suppl. Meth. 39. 1802 (Available from http://www.efloras.org/florataxon.aspx?flora_id=2andtaxon_id=106920)

12. Hamidun Bunawan. Siti Noraini Bunawan. Syarul Nataqain Baharum. The Red Butterfly Wing (Christia vespertilionis): A Promising Cancer Cure in Malaysia. International Journal of Pharmacy and Pharmaceutical Sciences. 2015; 7(8): 1-1.

13. Patil SG. Wagh AS. Pawara RC. Ambore SM. Standard tools for evaluation of herbal drugs: An overview. The Pharma Innovation. 2013; 2(9): 1-6.

14. Khan W. Mujum A. Shaikh T. Katekar SM. Tambe R. Rub RA. Pharmacognostic and preliminary phytochemical investigation of Salvadora persica Linn (Salvadoraceae). Research Journal of Pharmacognosy and Phytochemistry. 2010; 2(4): 319-23.

15. Mukherjee PK. Quality control and evaluation of herbal drugs: Evaluating natural products and traditional medicine. Elsevier; 2019 May 30.

16. Evans WC. Trease GE. A Textbook of Pharmacognosy. Baillière, Tindall and Cassell; The West African Medical Journal. Nigeria: University College. 1966.

17. Mahitha B. Archana P. Ebrahimzadeh MH. Srikanth K. Rajinikanth M. Ramaswamy N. In vitro antioxidant and pharmacognostic studies of leaf extracts of Cajanus cajan (L.) millsp. Indian Journal of Pharmaceutical Sciences. 2015; 77(2): 170. https://doi.org/10.4103/0250-474x.156555

18. Muthukumarasamy R. Kamaruddin A. Radhakrishnan S. Comparative evaluation of different extraction methods for antioxidant activity from Citrus hystrix peels. Drug Invention Today. 2018; 10(8): 1458-1462.

19. Neha B. Honey J. Ranjan B. Mukesh B. Pharmacognostical and preliminary phytochemical investigation of Acorus calamus linn. Asian Journal of Pharmaceutical Research. 2012; 2(1): 39-42.

20. Ranjith D. Fluorescence analysis and extractive values of herbal formulations used for wound healing activity in animals. J Med Plants Stud. 2018; 6(2): 189 - 192.

21. Rajabudeen E. Ganthi AS. Subramanian M. Pharmacognostical Studies on Tephrosia villosa (L.) Pers. (Fabaceae). Research Journal of Pharmacognosy and Phytochemistry. 2014; 6(4): 190-4.

Received on 07.02.2023 Modified on 19.07.2023

Research J. Pharm. and Tech 2024; 17(6):2452-2458.

DOI: 10.52711/0974-360X.2024.00383