Metabolite Profile Assessment and Phytochemical Comparative Studies of Ethanolic Extracts of Leaf and Flower Parts of Medicinal Plant Epiphyllum oxypetalum Lin. (Bramhakamal)

Raje Siddiraju Upendra¹, Arpitha C T², Sanjay Shrinivas Nagar³, R Karthik⁴

¹School of Electronics and Communication Engineering, REVA University, Rukmini Knowledge Park, Kattigenahalli, Yelahanka, Bengaluru - 560064, Karnataka, India.

²School of Applied Sciences, REVA University, Rukmini Knowledge Park, Kattigenahalli, Yelahanka,

Bengaluru - 560064, Karnataka, India.

³School of Applied Sciences, REVA University, Rukmini Knowledge Park, Kattigenahalli, Yelahanka,

Bengaluru - 560064, Karnataka, India.

⁴School of Electronics and Communication Engineering, REVA University, Rukmini Knowledge Park, Kattigenahalli, Yelahanka, Bengaluru - 560064, Karnataka, India.

*Corresponding Author E-mail: upendra.rs@reva.edu.in

ABSTRACT:

Epiphyllum oxypetalum, also known as Bramhakamal in India, belongs to the cactus family, Cactaceae, and holds significant cultural and religious value. This plant stands out as one of the highest cultivated species in its genus. While it has been acknowledged for its holy significance, its potential therapeutic properties have gained attention due to its limited exploration in phytochemical research. The present study aimed to assess the nutritive value, biotherapeutic potentials, and secondary metabolite outline of both leaf and flower parts of Epiphyllum oxypetalum. Nutritive analysis revealed essential components like proteins, lipids, and vitamins, including niacin. Further analysis of native extracts identified various secondary metabolites such as alkaloids, terpenoids, flavonoids, and glycosides. Antibacterial activity was observed in flower extracts, while fluorescent analysis of flower powder with different chemical reagents provided insights into its phytochemical characteristics. The focus extended to specific phytochemicals in leaf isolates using Gas Chromatography-Mass Spectrometry (GC-MS), uncovering the presence of eight medicinally active metabolites. Notably, Beta-Sitosterol emerged as the prominent compound (23.06%), known for its therapeutic applications in heart disease, rheumatoid arthritis, AIDS, and as an anticancer agent against cervical and colon cancer. This comprehensive study highlights the medicinal potential of Epiphyllum oxypetalum, emphasizing the elite presence of Beta-Sitosterol and elucidating the nutritive values of its leaf and flower parts. Further exploration, particularly through GC-MS analysis of flower extracts, could broaden our understanding of their pharmacological properties.

KEYWORDS: Epiphyllum oxypetalum, Leaf and Flower, Phytochemical constituents, Beta-sitosterol, GC-MS.

1. INTRODUCTION:

Epiphyllum oxypetalum belongs to the cactus species and is a prominent member of its genus, is widely cultivated, exclusively in regions with warmer climates such as parts of America, including California. It is also found naturally covering from Mexico to Venezuela and into Brazil. This variety of night-blooming Cereus derives its name from the Latin "Oxypetalum," referring to its sharply pointed petals. It is typically acknowledged as the Orchid cactus, Queen of night, or Lady of night, Dutchman's Pipe, Night-blooming Cereus, and locally as Wijaya Kusuma (Indonesian) and Nishagandhi (Hindi and Marathi).

Epiphyllum oxypetalum is mainly cultivated as an ornamental plant and is now being explored for its nutritious and antibacterial properties¹. Easily grown and fast-developing, Epiphyllum oxypetalum grows well in humus-rich compost with sufficient summer moisture, however, it requires protection from temperatures below 10 ^°C (50 ^°F) in winter. Epiphyllum oxypetalum also exhibits antidiabetic² and antiviral properties³. It adapts well to semi-shaded or fully sunny environments, with supplementary light exposure in early spring promoting flowering. The plant is characterized by erect, ascending, or spreading stems, this plant possesses abundant branching. Its primary stems are cylindrical and up to 2–6 m in length, woody at the base, whereas, secondary stems are flat and elliptical-acuminate, measuring around 30 cm long and 10–12 cm wide, with shallow to deeply crenate margins. Notably, its nocturnal flowers, measuring up to 30 cm in length and 12–17 cm in width, emit a highly fragrant aroma, with benzyl salicylate identified as a prime odour component. The exploration of therapeutic properties within plant extracts through chemical, pharmacological, and toxicological studies is a promising research area. Given the vast reservoir of pharmacologically active compounds found in the plant kingdom, there is increasing interest from pharmaceutical companies in identifying new bioactive substances derived from plants, leading to a proliferation of commercially available plant-derived medicines⁴.

1.1. Antibacterial Activity:

Beyond its aesthetic attraction, recent studies have revealed its capability as a source of bioactive compounds with notable antibacterial activity. The mode of action of antibacterial efficacies of flower and leaf organic extracts of the plant Epiphyllum oxypetalum was discussed in detail (Figure 1). It is estimated that plant extracts targeting sites that differ from those targeted by antibiotics will demonstrate activity against drug-resistant pathogens⁵.

These compounds have been employed to treat various microbial-related diseases. Alkaloids, phenolic compounds, tannins, and flavonoids, are the major bioactive compounds in these plants, which act as natural defenders against various pests and pathogens⁶. The utilization of plants in traditional health medicines is prevalent and vital for nearly 80% of the global population, especially in African, Asian, Middle Eastern countries, and Latin America, with stated minimum side effects⁷. These days, pharmaceutical companies have invested significant time and resources in developing therapeutics based on natural products derived from plants⁸. Mahmad et al. (2022) studied the antibacterial activity of the methanolic leaf extract of Epiphyllum oxypetalum against Candida albicans, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, and Staphylococcus epidermidis. The extracts exhibited antibacterial activity against both gram-positive and gram-negative bacteria. The biochemical content of the Epiphyllum oxypetalum leaf extract has the potential as an antibacterial, together against gram-negative and gram-positive bacteria, compared to antifungals⁷.

1.2. Phytochemical Constituents:

Phytochemicals were referred to as phytonutrients/phytoconstituents which are reported in diverse categories of plants and are utilized as essential components of both animal and human diet, where they have exhibited physiological properties. These Phytoconstituents work with fibers and nutrients to form a combined part of the defense system against different diseases and stress conditions. Phytochemicals are categorized into two groups i.e., primary, and secondary constituents. Primary components consist of common amino acids, chlorophyll, proteins, and sugars, whereas secondary constituents contain alkaloids, flavonoids, phenolic compounds, terpenoids, tannins, saponins, and so on. Research on Epiphyllum oxypetalum has uncovered the presence of various phytochemical constituents that contribute to its biological activities⁹. These constituents include alkaloids, flavonoids, phenolic compounds, glycosides, and polysaccharides. Studies have shown that these phytochemicals exhibit antioxidant, anti-inflammatory, antimicrobial, and anticancer properties¹⁰. The flower revealed the presence of several phytochemical composites such as alkaloids, amino acids, flavonoids, proteins, saponins, terpenoids, and tannins. The crude ethanol extract characterized through GC-MS analysis exposed the presence of flavonoid compound [7-hydroxy-3(1,1-dimethyl prop-2enyl) coumarin and other compounds like Hexadecenoic acid, Nonadecanoic acid, and Oleic acid, which were also stated in the leaf extract¹¹.

2. METHODOLOGY:

2.1. Preparation of Plant Crude Extract:

Fresh leaf and flower from the plant Epiphyllum oxypetallum were procured locally from Lalbagh Botanical Garden, which was taxonomically identified by the Department of Botany, IISC Bangalore for the process of extraction of bioactive molecules. Both leaf and flower retrieved from the plant were dehydrated and converted into powder for further studies. From the total powdered plant material (leaf and flower) 20 gm was weighed, which was separated in a 200 ml conical flask containing 100 ml of water as the solvent. The head part of a 200 ml conical flask containing plant sample and solvent was enclosed with aluminum foil and both components were mixed properly in the reciprocating shaker at the speed of 150 rev/min for the duration of 24 hr, which ensured proper mixing of powdered plant material with solvent. Plant extract produced in a reciprocating shaker was filtered with the help of muslin cloth along with Whatman no 1 filter paper to ensure the removal of the impurities from the plant extract. After filtering out the impurities, the solvent present in the extract was separated through an evaporation process performed at 50 ^°C inside the water bath apparatus. The extract obtained after the removal of the solvent was dried at room temperature. The dried form of the pure extract was stored in a sterile vacuum pouch and used for the extraction of bioactive molecules¹².

2.2. Nutritive Analysis of Epiphyllum oxypetallum flower:

2.2.1. Estimation of proteins - Xanthoprotein Test:

Pure extract (1 ml) and Distilled Water (DW) (1 ml) (Control) were collected in test tubes for protein prediction. 0.5 ml of Nitric acid was added into both test tubes and mixed properly. Exothermic reaction in test tubes caused due to the addition of nitric acid was controlled through tap water, which was followed with the addition of 1 ml of 40% Sodium hydroxide to both test tubes. The test tube displaying yellow colour provided qualitative proof of protein content in sample ¹³.

2.2.2. Estimation of Starch – Iodine Test:

A mixture consisting of dilute HCl, iodine solution, and the extract was prepared in the test tube resulting in the appearance of blue colour. Test tube contents exhibiting blue colour were subjected to heat till the disappearance of blue color followed by cooling of the test tube. The resurgence of blue solution indicated the presence of starch in the tested extract¹³.

2.2.3. Determination of Lipids:

Extract (0.2 gm), solvent (50 ml), and phenolphthalein (1 ml) were taken in a conical flask and titrated against 0.1 N potassium hydroxide with constant pink colour as titration endpoint. At the end of titration process, the difference between final and initial values of the burette was used for the detection of free fatty acids in the extract tested¹⁴.

2.2.4. Estimation of Reducing sugars:

Plant extract and DW were mixed in the test tube and resulting mixture was filtered. Filtrate was combined with a few drops of Fehling’s solution in water bath, where the appearance of an orange precipitate signifies the presence of reducing sugars in the plant extract tested¹⁵.

2.2.5. Vitamin estimation (Niacin (B3)): A mixture comprising powdered extract (5 gm) and sulphuric acid (30 ml) was processed under steam for 30 minutes. The volume of processed mixture was elevated from 30 to 50 ml through the addition of DW and resulting slurry was filtered through Whatman no.1 filter paper. 60% of basic lead acetate was added to the filtrate and the pH of the resulting mixture was adjusted to 9.5 value using 10 N sodium hydroxide and pH meter. A mixture of 9.5 pH was processed through centrifugation, followed by the addition of 2 ml conc. sulphuric acid and the entire mixture was kept still for 1 hour. The mixture with sulphuric acid was subjected to second centrifugation followed by the addition of 40% ZnSO4(H2O)x (5ml) and 10N sodium hydroxide to maintain the pH value of 8.4. A mixture of 8.4 pH was processed for third centrifugation and pH of the supernatant was adjusted to 7.0 and used as the final mixture for the vitamin test. Two main raw materials required for vitamin estimation test are cyanogen bromide solution and standard solution. 1ml of standard solution contains 100mg of niacin. Five tests in series were performed with increasing values of standard niacin and extract (7.0 pH) from 0.1 ml to 0.5 ml separately. Volume of 10 test tubes (5 standard and 5 extracts) were increased up to 6ml with the addition of DW, which was followed by the addition of 3 ml of cyanogen bromide. Contents present in test tubes were maintained in an idle position for 10 minutes, which was followed by the addition of 4% Aniline (1ml) to the test tubes. Test tubes displaying yellow colour were evaluated with a calorimeter at 520 nm. A standard graph was generated to calculate niacin content of extract tested¹⁵.

2.3. Preliminary phytochemical analysis: Analysis of petroleum ether, chloroform, and ethanol extracts of Eryngium foetidum was carried out to identify the secondary metabolites¹⁶.

2.3.1. Alkaloid test: 1gm extract was dissolved in DW and Wagner’s reagent. Display of reddish-brown precipitate indicated the presence of alkaloid¹⁷.

2.3.2. Flavonoid test: Addition of 10 ml DW and a few drops of 20% NaOH to extract (1 gm) produced a yellow solution, which changed into a colourless solution due to nitric acid addition and indicated flavanoid pressence¹⁶.

2.3.3. saponins: About 0.2 g of the extract was shaken with 5ml of distilled water and then heated to a boil. Frothing (appearance of creamy miss of small bubbles) shows the presence of saponins¹⁷.

2.3.4. Tannins test: Extract in lesser quantity was heated with water in a water bath and filtered to obtain the filtrate, which was combined with ferric chloride yielding a dark green solution indicating the presence of tannins¹⁷.

2.3.5. Glycoside test: 0.5 ml extract was taken into a test tube containing sulphuric acid. The reaction of sulphuric acid with extract displayed a bluish-green color as the extract's upper layer and indicated the presence of glycosides¹⁷.

2.3.6. Terpenoid test: 0.2 gm extract was mixed with chloroform (2 ml) and conc H₂S0₄ (3 ml) resulting in reddish brown colour in between two extract layers and thus indicating the presence of terpenoids¹⁷.

2.3.7. Phlobatanin test: Extract was dissolved in DW and the resulting slurry was filtered to obtain a filtrate. Filtrate along with 2% HCl and heated at high temperature resulting in the creation of red precipitate, thus indicating phlobatanins presence¹⁸.

2.3.8. Carotenoid test: Extract was mined with 10ml chloroform as a solvent with constant shaking. The mixture obtained was filtered and 85% H₂SO₄ was added to the filtrate. Display of blue colour in between two layers proved carotenoid existence in extract¹⁸.

2.3.9. Quinones test: The addition of dil NaOH to plant extract (1 ml) resulted in a blue-green solution and thus revealed quinones existence in the extract studied¹⁹.

2.3.10. Coumarin test: The addition of 10% NaOH to plant extract (1 ml) resulted in a yellow solution and thus revealed coumarin existence in the extract studied¹⁹.

2.4. Phenolic compounds test: Extract (50 mg) was mixed with DW (5 ml), 5% iron (III) chloride (3-4 drops), and potassium ferro cyanide (1ml). The mixture displayed dark green colour and justified the existence of phenolic compounds in the extract studied²⁰.

2.4.1. Resin test: Extract was mixed in DW and studied for its turbidity.

2.4.2. Acidic Compound test: Sodium bicarbonate mixed with extract was studied for its foaming.

2.5. Antibacterial activity:

2.5.1. Collection of microorganisms and preparation of media: Stock cultures were obtained from MTCC including bacteria such as Escherichia coli (MTCC 443) and Bacillus subtilis (MTCC 9003). Growth media employed in the present study were nutrient agar and nutrient broth media adjusted to pH 7.4 and media sterilization was performed through an autoclave apparatus at a temperature of 121 ^°C for a duration of 15 minutes²¹.

2.5.2. Preparation of Inoculum: Organisms of the primary plate were subcultured onto nutrient agar for studying the viability of microbes incubated on subculture. Seed microbes were incubated onto nutrient agar slopes at 4 ^°C temperature and later sub-incubated in the nutrient broth at 37 ^°C temperature before the antibacterial test²⁰.

2.5.3. Agar Disc Diffusion (ADD): ADD was applied to investigate the antibacterial properties of the extract studied. Microbes present in the sample were spread across the petri plates containing nutrient agar. Filter discs (6 mm) drenched with extract were placed onto the surface of inoculated or agar plates and allowed to dehydrate for 15 minutes. Dry agar plates were incubated at 37 ^°C temperature for 24 hours. After 24 hours the width of inhibition zones was measured in mm²¹.

2.6. GC-MS: GC-MS is used for the analysis of unknown samples. The extracted sample to be analyzed was injected into the inlet of GC, where the sample was converted into vapour form and spread on the surface of the chromatographic column with helium gas as the mobile phase. Extract flows through the stationary phase leading to the charecterization of molecules based on the binding affinity between extract and stationary. Compounds displaying lower binding affinity with the column are eluted first and compounds with more binding affinity with the column are eluted later. The latter segment of the column proceeds through the heated transfer line, culminating at the ion source entrance. Here, all eluted compounds are transformed into ions²³.

3. RESULTS AND DISCUSSION:

3.1. Crude Extract:

Native extraction of leaf and flower samples of Epiphyllum oxypetalum yielded significant results, revealing the existence of primary and secondary metabolites. The extraction process effectively isolated a diverse array of phytochemicals from both plant parts. In the leaf extraction, high concentrations of glycosides, phenolic compounds, saponins, resins, tannins, and terpenoids were observed. These compounds are known for their antioxidant properties, which contribute to the plant's ability to combat oxidative stress and protect against various diseases. Furthermore, the flower extraction exhibited notable levels of alkaloids, flavonoids, phenolic compounds, terpenoids, and glycosides. Alkaloids, in particular, play a crucial role in pharmacological activities due to their diverse biological effects, including antibacterial features. Terpenoids and glycosides signify the therapeutic value of plants, such as antimicrobial, anticancer, and antioxidant activities. The findings from this native extraction highlight the pharmacological potential of Epiphyllum oxypetalum leaf and flower extracts. Further studies can reveal the bioactive compounds with active pharmacological activities.

Native Extraction of Leaf:

Fresh leaves and flowers of the plant obtained from the botanical garden were dried and the dried leaf and flower were converted into powder. Powdered leaf and flower were mixed separately with solvent and dried to obtain the final leaf and flower powder for extraction of bioactive components as discussed in Figure 2.

Table 2: Phytochemical analysis

S. No	Test	Flower Extract
1.	Phenolic compounds	Positive
2.	Alkaloids	Positive
3.	Glycosides	Positive
4.	Quinones	Negative
5.	Coumarins	Positive
6.	Saponins	Negative
7.	Resins	Positive
8.	Terpenoids	Positive
9.	Acidic compounds	Negative
10	Phlobatanins	Negative
11.	Flavonoids	Positive
12.	Carotenoids	Negative
13.	Tannins	Negative
14.	Proteins	Positive
15.	Starch	Negative
16.	Reducing sugar	Negative
17.	Lipids	Positive

3.3. Antibacterial activity of flower extracts:

The antibacterial property of Epiphyllum oxypetallum flower extract was tested against E. coli and Bacillus subtilis. Zones of inhibition were observed around discs inoculated with flower extract, indicating antibacterial properties. Figure 3A shows the antibacterial properties of the flower derivatives or extract against bacteria E.coli and Figure 3B shows the antibacterial properties of flower derivative against bacteria Bacillus subtilis.

Figure 3. Antibacterial activity of the bioactive molecules extracted from the flower of plant Epiphyllum oxypetallum against the microorganism E.coli MTCC443 . (3A) and Bacillus subtilis MTCC9003 (3B). The formation of the zone around Ab (Antibody) disc indicates the antibacterial property of the extract studied.

3.4. Fluorescent analysis of Epiphyllum Oxypetallum powder on treatment with different reagents:

Fluorescence studies were conducted on Epiphyllum Oxypetallum flower extract in powdered form applying numerous reagents under UV and visible light conditions (Table 3) revealing distinct characteristics. Fluorescence, an essential phenomenon observed in plant materials, showcases the presence of different chemical constituents. While some constituents exhibit visible fluorescence in daylight, others require ultraviolet light to induce fluorescence. Additionally, substances that are not inherently fluorescent can be transformed into fluorescent derivatives through the application of specific reagents. Consequently, qualitative assessment of crude drugs often relies on fluorescence studies, which serve as a crucial parameter in pharmacognostical evaluation. Figure 4 shows the fluorescent analysis of flower reagents.

3.5. Comparative analysis of the profile of the bioactive compounds studied for both leaf and flower derivatives or extracts of Epiphyllum oxypetalum:

The comparison of metabolite profiles between leaf and flower extracts of Epiphyllum oxypetalum revealed distinct differences in their chemical composition. Saponins, alkaloids, and flavonoids show notable variations between the two extracts, with saponins, alkaloids, and flavonoids being present in the leaf extract but absent in the flower extract. This suggests that these compounds may play specific roles in the physiological functions of the plant parts. glycosides, resins, and terpenoids, these metabolites are present in both leaf and flower extracts, indicating their widespread distribution throughout the plant. Acidic compounds and Phlobatanins, these compounds are absent in both extracts, suggesting their minimal or negligible presence in Epiphyllum oxypetalum. Phenolic compounds were present in both extracts, they contribute to various biological activities and may contribute to the therapeutic potential of the plant. Tannins are absent in the flower extract but present in the leaf extract. This difference may influence the medicinal or nutritional properties associated with each plant part. Differences in the bioactive molecule composition in both leaf and flower derivatives are described in Table 4.

Table 3: Fluorescent analysis of Epiphyllum Oxypetallum flower

S. No	Reagents	Daylight	UV 254 nm
1	Powder as such	Light brown	Dark green
2	Ferric chloride	Brown	Green
3	50% Nitric acid	Yellow	Dark Green
4	Sulphuric acid	Maroon	Dark brown
5	NaOH	Beige	Brown
6	25 % Ammonia	Brown	Brownish green
7	Iodine	Red	Black

Table 4 Metabolite Comparison of Leaf and Flower

Test	Flower Extract	Leaf Extract
Alkaloids	+	-
Glycosides	+	+
Resins	+	+
Flavonoids	+	-
Terpenoids	+	+
Saponins	-	+
Acidic compounds	-	-
Phlobatanins	-	-
Phenolic compounds	+	+
Tannins	-	+

3.6. Gas Chromatography-Mass Spectrometry (GC-MS) study:

GC-MS study of the target sample identified several bioactive compounds, including Alpha Campholenal, p-Methoxycinnamic Acid, 2-Ethylhexyl-4-Methoxy Cinnamate, 4,8,12,10-Tetra Methyl Heptadecan-4-Olide, Trans-Geranylgeraniol, Spinacene, Alpha Tocopherol, and Beta Sitosterol. These compounds exhibit diverse properties such as antioxidant, photoprotective, and cholesterol-lowering effects, highlighting the sample's potential in various applications including pharmaceuticals, and cosmetics. Further research is needed to explore the specific biological activities and potential benefits associated with these compounds. GC-MS analysis of leaf is presented in Table 5.

Table 5: GC-MS Analysis of Leaf

Sl. No.	Retention Time	Compound Name
1	7.029	Alpha campholenal
2	13.37	p-methoxy cinnamic acid
3	14.85	2-Ethylhexyl-4-methoxy cinnamte
4	15.01	4,8,12,10-tetra methyl heptadecan-4-olide
5	18.72	Trans-Geranylgeraniol
6	18.74 / 18.75	Spinacene
7	21.38	Alpha tocopherol
8	23.712/23.726/23.722/23.73	Beta sitosterol

GC-MS graph was performed for both good and raw samples of Bramhakamala with both petroleum ether and acetone as organic solvents to analyze the bioactive molecules of the plant extract. Six distinct peaks in the graph led to the discovery of the compounds Beta-sitosterol, Spinacene, and 2-ethylhexyl-4-methoxy cinnamate (Figures 5A, 5C, and 5E). Elven distinct peaks were identified that led to the discovery of Alpha campholenal, p-methoxycinnamic acid, 4,8,12,10-tetra methyl heptadecan-4-olide, Trans-Geranylgeraniol, Alpha tocopherol (figure 5B, 5D and 5F).

DISCUSSION:

Epiphyllum oxypetalum belongs Catus species and is a prominent member of its genus, is widely cultivated, exclusively in regions with warmer climates such as parts of America, including California. Given the vast reservoir of pharmacologically active compounds found in the plant kingdom, there is increasing interest from pharmaceutical companies in identifying new bioactive substances derived from plants, leading to a proliferation of commercially available plant-derived medicines. Bioactive molecules can also be extracted from agricultural plants to treat various diseases²¹and pandemic diseases such as COVID-19. Many authors have studied bioactive molecules in the plant Epiphyllum oxypetalum of methanol root extract. Extract preparation was performed with the help of standard laboratory protocol to study the bioactive components in the root of the plant Epiphyllum oxypetalum²². The team discovered the presence of various bioactive molecules such as protein, fats, crude fiber, and carbohydrates in the root of the target plant. A detailed study can be performed to study the presence of specific bioactive components in the root of the target plant. The study can also be extended to other parts of the target plant such as flower and leaf²³. Very less studies have been conducted in the field of analyzing the bioactive components of the plant Epiphyllum oxypetalum for various biological activities. Hence there is a need to perform a study to identify potential bioactive molecules present in the leaf and flower of the plant Epiphyllum oxypetalum. The present study investigated the antibacterial properties and nutritional composition of leaf and flower extracts of Epiphyllum oxypetalum, shedding light on its potential therapeutic applications and nutritional benefits. The study revealed significant antibacterial activity of both leaf and flower extracts, suggesting their potential utilization in managing bacterial diseases in human systems. Additionally, the nutritional analysis of flower extracts indicated the presence of proteins, fatty acids, and vitamins, which are essential components for human health. However, carbohydrates were found to be absent in the flower extracts. The presence of these nutrients adds to the overall positive attributes of the plant's flower. Furthermore, the preliminary phytochemical screening of the plant's flower extracts detected the presence of alkaloids, flavonoids, terpenoids, phenolic compounds, and glycosides, while reducing sugars, saponins, and tannins were absent. These secondary metabolites are known for their medicinal properties and may contribute to the plant's therapeutic effects on human physiology. Moreover, the GC-MS²⁴ analysis of Epiphyllum oxypetalum leaf extracts identified seven bioactive components, with beta-sitosterol being the prominent one. Beta-sitosterol is recognized for its potential therapeutic applications in treating cancer, colon, HIV, and influenza. The study also highlighted the importance of fluorescence as an indicator of various chemical constituents present in plant material²⁵. Further research could focus on detecting specific secondary metabolites present in the flower extracts using advanced techniques such as GC-MS. The findings of this study suggest that Epiphyllum oxypetalum possesses significant antibacterial properties and nutritional value, making it promising for the management of bacterial diseases. The presence of various bioactive components further supports its potential therapeutic applications. Future studies could delve deeper into identifying specific metabolites and elucidating their role in the development of novel therapeutic agents derived from Epiphyllum oxypetalum.

CONCLUSION:

The present study investigated the antibacterial properties and nutritional composition of leaf and flower extracts of Epiphyllum oxypetalum, shedding light on its potential therapeutic applications and nutritional benefits. The study revealed significant antibacterial activity of both leaf and flower extracts, suggesting their potential utilization in managing bacterial diseases in human systems. Additionally, the nutritional analysis of flower extracts indicated the presence of proteins, fatty acids, and vitamins, which are essential components for human health. However, carbohydrates were found to be absent in the flower extracts. Future studies could delve deeper into identifying specific secondary metabolites and elucidating their pharmacological activities, paving the way for the development of novel therapeutic agents derived from Epiphyllum oxypetalum.

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

ACKNOWLEDGMENTS:

The authors would like to thank REVA University for their kind support in conducting research work.

FUNDING SUPPORT:

The authors have not received any funding support for the research work performed.

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Received on 08.08.2024 Revised on 17.01.2025

Accepted on 01.04.2025 Published on 01.12.2025

Available online from December 06, 2025

Research J. Pharmacy and Technology. 2025;18(12):5682-5692.

DOI: 10.52711/0974-360X.2025.00821

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

S. No.	Std. Niacin(ml)	DW (ml)	Cyanogen bromide (ml)	Shake For 10min	4% Aniline(ml)	Yellow should be developed after 5 min	OD 420nm
S1	0.1	5.9	3		1		0.09
S2	0.2	5.8	3		1		0.18
S3	0.3	5.7	3		1		0.26
S4	0.4	5.6	3		1		0.39
S5	0.5	5.5	3		1		0.40
T1	0.2	5.8	3		1		0.14
T2	0.5	5.5	3		1		0.42