INTRODUCTION

Medicinal plants are an essential source of several bioactive compounds, which have been used to treat several diseases caused in human beings. The World Health Organization (WHO) estimates that 80% of developing countries rely on therapeutic drugs (Vital & Rivera, 2009). Their low cost, easy accessibility and the search on medicinal plants have led to the discovery of novel therapeutic drugs against diseases.

Gas chromatography is an exciting tool used nowadays to determine new bioactive compounds from critical medicinal plants, which interest is discovering novel drugs. The identified compounds are used in cosmetics, drugs, pharmaceuticals, the food industry and forensic applications (Uma, Prabhakar, Rajendran, & Lakshmi, 2009). Identifying bioactive compounds from different plant parts leads to an additional investigation of pharmacological and biological studies (Farid et al., 2015; Momin et al., 2014). Different potent active compounds in the various plant extracts can treat various human diseases (Konappa et al., 2020). Most of the world’s population depends on plant-derived medicines, which has drawn the interest of researchers to invent new drugs. (Iwu, Duncan, & Okunji, 1999). The infections caused by microbes threaten a major health hazard that is around 25% of all the deaths among the countries (Priyanka, Kumar, Bankar, & Karthik, 2015). Natural products have been mainly a rich source of anti-infective agents. Many researchers have screened the in vitro crude extracts from plants with a history of use in folk medicine have been screened for antibacterial activity by many researchers (Cushnie & Lamb, 2005).

According to these reports, several pharmacological studies have been demonstrated using various plant parts of M. calabura. The M. calabura is a beautiful flowering shrub that belongs to the Muntingaceae family. The secondary metabolites of plants such as alkaloids, flavonoids, phenols, tannins and saponins were reported in the leaf extracts of M. calabura. (Buhian, Rubio, Valle, & Puzon, 2016). These active compounds are essential in the food and research industry (Leema & Prakash, 2019). The leaf and root parts of M. calabura have high anti-inflammatory, antiulcer, antibacterial, antipyretic, and antioxidant activities (Krishnaveni, Raginabanu, Kalaivani, & Krishnakumari, 2015; Sufian, Ramasamy, Ahmat, Zakaria, & Yusof, 2013; Zakaria et al., 2007; Zakaria et al., 2011; Zakaria et al., 2014).

Thus, the present study evaluates the presence of bioactive compounds present in methanolic extracts of M. calabura leaf and root parts using GC-MS analysis and evaluations of their antibacterial activity by using the agar well diffusion method.

MATERIALS AND METHOD

Plant material collection and sample preparation

Fresh M. calabura leaves and roots were collected from the greenhouse at (18°01'35.1"N 79°33'31.7"E). The collected plant materials were thoroughly washed under running tap water to remove impurities. These were dried for 12 days in the shade at room temperature (27 ± 2 °C) and ground into a fine powder. Five gram of plant materials were mixed with 50 mL of methanol and incubated in an orbital shaker for 48 hours at 130 rpm. Whatman No. 1 filter paper was used to filter the supernatant. The supernatants were evaporated at room temperature to obtain a crude extract, then subjected to GC-MS analysis and anti-microbial activity testing.

GC-MS instrumentation

M. calabura leaf and root methanol extracts were GC-MS analysed using a Perkin-Elmer GC system coupled with a silica capillary column 30m x 0.25mm ID x 0.25m df equipped with Elite-5MS (5 % diphenyl and 95 % dimethyl polysiloxane). Helium was used as a gaseous carrier in a 1:1 mg/minute ratio, with an injector volume of 2μl and a temperature of 250 °C and ionization at 70-eV. The oven temperature began at 110 °C and ended at 280 °C. The extraction time was 29 minutes for M. calabura leaf extracts and 13 minutes for root methanolic extracts. The total peak area of M. calabura was used to calculate the percentage of each bioactive compound in leaf and root extract. The Sophisticated Analytical Instrument Facility, IIT Bombay, performed the GC-MS analysis.

Microbial cultures and culture conditions

Muntingia calabura methanolic leaf and root extracts were tested against gram-negative and gram-positive bacteria. Escherichia coli, Proteus vulgaris, Bacillus sphericus, and Pseudomonas fluroscens were used as test organisms. Preparation of 24 hr old bacterial strains: inoculate nutrient broth with original cultures and incubate overnight at 37 °C. The antibacterial activity was tested using the agar well diffusion method with streptomycin (10 µg/mL) as the standard. The media was prepared and poured 15 mL into a petri dish for 5 minutes to solidify. The inoculums were swabbed uniformly over the media and dried. The standard (10 µg/mL) and three different leaf and root extract concentrations (45, 60, and 75 µg/mL) were incubated overnight at 37 °C in each well. After incubation, zones emerged, and the proportion of inhibition was scaled in millimetres.

RESULTS AND DISCUSSION

M. calabura phytochemical components were detected in methanolic leaf and root extracts. The percentage of the compounds was estimated using the retention time (RT) and peak area.

GC-MS analysis of leaf methanolic extract

The bioactive molecules found in methanolic leaf extracts were thirty-eight compounds. Among the thirty-eight compounds, the first compound identified with less retention time (4.85 min) was 1,3-dihydroxypropan-2-one and the peak area (0.24%), whereas the last compound with the longest retention time (33.68 min) was 3-Hydropregn-5-en-20-one hydrazone and the peak area (0.33%). Many of these compounds possess various pharmacological activities. The remaining phytochemical compounds are as follows: 1-(6-oxabicyclo[3.1.0]hexan-1-yl)ethan-1-one (or) 6-oxabicyclo(3.1.0)hexan-3-one, a phytoconstituent of methanolic leaf and root extracts of pili and safedshatavar (Asparagus racemosus), known to possess antioxidant activity (Banakar & Jayaraj, 2017). The 3,5-Dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one reported antimicrobial, automatic nerve, and antioxidant properties in ethanolic stem extracts of an important medicinal plant Waltheria indica L (Ashwathanarayana & Naika, 2018). 2,3-dihydro-1-benzofuran, a bioactive compound, possesses anti-inflammatory activity in crude extracts of leaf and flower of Pavetta crassicaulis Bremek (Correa et al., 2017). Decan-3-yl acetate, reported to possess activities like asthma, anticancer, anti-inflammatory activities and Histamine H3 and H4 receptors, have been discovered drugs for Parkinson’s and Alzheimer’s disease (Tatipamula et al., 2019). The 1,2,3-benzenetriol (Pyrogallol), identified as a phytochemical compound that possesses anticancer activity in ethanolic extract of Clathria baltica (Sukprasert, Pansuksan, & Sriyakul, 2020). The compound (1R,2S,3R,4S)-cyclopentane1,2,3,4-tetrol, known to possess antiviral activity in Lysiphyllum strychnifolium plant extracts that treat against influenza virus (Rubaye, Hameed, & Kamal, 2018). The compound, 4-[(1E)-3-hydroxyprop-1-en-1-yl]-2-methoxyphenol have multiple biological properties like anti-hyperglycemic, hepatoprotective, anti-obesity and antibiotic activities in root extracts of Eclipta alba (L) (Naik, Gurushanthaiah, Kavimani, & Mahesh, 2019). More recently, it was reported that the compound (2E)-3,7,11,15-tetramethyl-2-hexadecen-1-ol has antifungal and antibacterial activity in leaf extracts Rhizophora apiculata (Lakshmanan et al., 2019).

 Hexadecanoic acid, a phytochemical compound, has properties like strong antimicrobial, anti-inflammatory activities and acts against arthritis in medicinal plants like Albizia adianthifolia, Pterocarpus angolensis and in some plants against food-borne pathogens (Abubakar & Majinda, 2016; Preethi, Devanathan, & Loganathan, 2010). In Cycas revolute, a medicinal plant is known to have antioxidant properties in the 1–hentetracontanol bioactive molecule (Olabisi & Olubunmi, 2019). The (9Z,12E)-octadeca-9,12-dienoic acid and (9Z)-octadec-9-enoic acid have phytochemical properties like hypercholesterolemia, anti-acne, anti-cancer, anti-bacterial, and diabetes mellitus in methanolic leaf extract Crateva adansonii DC and root extract of Jatropha pelargoniifolia Courb (Christiana et al., 2014). Another bioactive molecule, 3-[(4-ter-butylcyclohexyl)oxy]-3-phenyl-2-benzofuran-1(3H)-one is identified to synthesize green synthetic particles using ferrous and nickel nanoparticles by leaf extract of eucalyptus (Weng, Guo, Luo, & Chen, 2017). 5-hydroxy2-phenyl-4–chromanone is one compound that possesses Helicobacter pylori and Gastric antisecretory activity (Das, Tiwari, & Shrivastava, 2010). 1-tetradecene,2–decyl reported to have phytochemical properties like an anti-inflammatory in leaf extracts of Azima tetracantha Lam (Jose & Panneerselvam, 2019) and antioxidant activity in leaf and stem of Strobilanthes species (Fernandes & Krishnan, 2019). Recently, the anti-inflammatory activity of (2E)-1,3-diphenylprop-2-en-1-one has been reported by (Landim et al., 2019) in Lonchocarpus cultratus.

Table 1

GC-MS of bioactive compounds present in the methanolic extracts ofleaves derived from in vivo grownplants of Muntingia calabura L

S No

RT

Compound name

Formulae

Mwt

Area

Biological activity

1

4.85

1,3-dihydroxypropan-2-one

C3H6O3

90

0.24%

Ant oxidative (Leema et al., 2019)

2

8.49

1-(6-oxabicyclo[3.1.0]hexan-1- yl)ethan-1-one (or) 6-oxabicyclo(3.1.0)hexan-3-one

C7H10O2

126

0.25%

Antimicrobial, automatic nerve activity and antioxidant (Banakar et al., 2017)

3

9.68

3,5-Dihydroxy-6-methyl-2,3- dihydro-4H-pyran-4-one

C6H8O4

144

0.16%

Anti-inflammatory (Ashwathanarayana et al., 2018)

4

11.20

2,3-dihydro-1-benzofuran

C8H8O

120

0.3%

Asthma, Anticancer, Parkinson’s and Alzheimer’s disease (Correa et al., 2017)

5

11.56

decan-3-yl acetate

C14H28O2

228

0.07%

Anticancer (Tatipamula et al., 2019)

6

14.20

benzene-1,2,3-triol (Pyrogallol)

C6H6O3

126

0.17%

Antiviral (Sukprasert et al., 2020)

7

15.22

-D-Glucopyranosyl-(1->3)-β-Dfructofuranosyl β-Dglucopyranoside (or) D-(+)-Melezitose

C18H32O16

504

0.03%

Nutritional Anemia (Wei et al., 2020)

8

15.81

(1R,2S,3R,4S)-cyclopentane1,2,3,4-tetrol (or) )-cyclopentane1,2,3,4-tetrola,4b,3b,2a(1

C5H10O4

134

0.44%

Antibacterial and antifungal (Rubaye et al., 2018)

9

18.02

4-(ethoxymethyl)-2-methoxyphenol

C10H14O3

183

0.10%

No activity

10

19.30

4-[(1 E)-3-hydroxyprop-1-en-1- yl]-2-methoxyphenol

C10H12O3

180

0.29%

Antihyperglycemic, hepatoprotective, Immunodulatory, Antiviral, Antifungal, Analgesic (Naik et al., 2019)

11

21.14

(2E)-3,7,11,15-tetramethyl-2- hexadecen-1-ol

C20H40O

296

0.13%

Antibacterial and antifungal (Lakshmanan et al., 2019)

12

22.35

hexadecanoic acid

C16H32O2

256

0.35%

Anti-inflammatory and Arthritis (Abubakar et al., 2016)

13

24.35

1 –hentetracontanol

C41H84O

592

0.07%

Antioxidant (Olabisi et al., 2019)

14

24.75

(9Z,12E)-octadeca-9,12-dienoic acid

C18H32O2

280

0.74%

Hypocholesterolemic, Anti-acne (Christiana et al., 2019)

15

24.97

(9Z)-octadec-9-enoic acid Or Oleic acid

C18H34O2

282

0.29%

Diabetes mellitus (Aati, El-Gamal, & Kayser, 2019)

16

26.77

N'-(4-tert-butylcyclohexylidene)- 4-methylbenzene-1- sulfonohydrazide

C17H26N2O2S

322

0.08%

No activity

17

27.22

4,8,12,16, tetramethylheptadecan-4-olide or 5-Methyl-5-(4,8,12- trimethyltridecyl)dihydro2(3H)-furanone

C21H40O2

324

0.16%

No activity

18

27.45

2H-1-Benzopyran-7 Ol,3,4-dihydro-5-methoxy-6-methyl 2-phenyl

C17H18O3

270

0.15%

No activity

19

27.61

3-[(4-ter-butylcyclohexyl)oxy]- 3-phenyl-2-benzofuran-1(3H)-one

C24H28O3

364

0.06%

Green synthetic particles (Weng et al., 2017)

20

27.92

5- hydroxy 2 - phenyl - 4 – chromanone

C15H12O3

240

1.24%

Helicobacter pylori and Gastric antisecretory activity (Das et al., 2010)

21

28.46

(2Z)-2-{[5-(methoxycarbonyl)-3,4- dimethyl-1H-pyrrol-2- yl]methylidene}-3,4,5-trimethyl2H-pyrrol-1-iumbromid

C17H23BrN2O2

366

0.47%

No activity

22

28.64

1-tetradecene,2–decyl

C24H48

336

0.02%

Anti-inflammatory, Arthritis and antioxidant (Fernandes et al., 2019)

23

28.92

4H-1-benzopyran-4-one,2,3-dihydro-5,7-dihydroxy-2-phenyl(3)

C15H12O4

256

2.11%

No activity

24

29.12

1,2-bis-[2-[4-cyclohepta-2,4,6-trienyl-phenoxy]-ethoxy]-ethane

C32H34O4

482

1.20%

No activity

25

29.24

(2E)-1,3-diphenylprop-2-en-1-one

C15H12O3

240

2.48%

Anti-proliferative (Landim et al., 2019)

26

29.56

2-{3 -[(E) - 2 -(1H -indol - 3 - yl)ethenyl]-1,2,4-oxadiazol-5- yl}phenol -

C18H13N3O2

303

5.78%

Antioxidant and Antitumor (Kopoytkoba & Caxho, 2019).

27

29.92

5 -hydroxy - 7 -methoxy - 2 -phenyl - 4H-1-benzopyran-4-one

C16H12O4

268

0.63%

Anticancer, anti-prostate cancer and anti-inflammatory activities (Wang et al., 2019).

28

30.01

(2S)-5,7-dihydroxy-2-(4- hydroxyphenyl)-2,3-dihydro-4H-1- benzopyran-4-one

C15H12O5

272

2.08%

Anti-tuberculosis, Anticancer (Sahu, Mishra, Chandra, Nirala, & Bhadauria, 2020).

29

30.32

7-hydroxy-3-methoxy-2-(4- methoxyphenyl)-4 H-1- benzopyran-4-one

C17H14O5

298

2.57%

Antioxidant (Suttiarporn, Sookwong, & Mahatheeranont, 2016)

30

30.44

4(1,1-Dimethylaaly)9-methoxy7H-furo (3,2-g)(1) benzopyrn-7-one

C17H16O4

284

0.52%

No activity

31

30.83

2,2' -((1E,1'E) -(1,4 - phenylenebis(azanylylidene))bis(m ethanylylidene))diphenol

C20H16N2O2

316

1.10%

Anti-tumour and Cancer phototherapy (Kopoytkoba et al., 2019)

32

31.29

5,7-dihydroxy - 2 -(4 - hydroxyphenyl)-6-methoxy-4Hchromen-4-one

C16H12O6

300

2.46%

Antioxidant (Karker, Kumar, & S, 2019; Mu et al., 2009; Yausheva et al., 2019)

33

31.67

2,3,7,8-tetramethoxy-5,11-dihydro10H-dibenzo[a,d][7]annulen-10- one

C19H20O5

328

1.08%

Arthritis, Anti-inflammatory (Fernandes et al., 2019)

34

32.17

1-(4-methylphenyl)anthracene - 9,10-dione

C21H14O2

298

2.15%

Anticancer (Chu et al., 2019)

35

32.78

8,9-dihydro-aflatoxin B1

C17H14O6

314

0.66%

Antimycotoxin (Pok et al., 2020)

36

33.15

1-Docosene

C22H44

308

0.74%

Anticancer (Swantara, Rita, Suartha, & Agustina, 2019)

37

33.45

3,6,8-trimethoxy-5,7-dimethyl-2- phenyl-4H-1-benzopyran-4-one

C18H16O7

344

0.09%

No activity

38

33.68

3-Hydropregn-5-en-20-one hydrazone

C21H34N2O

330

0.33%

No activity

Table 2

GC-MS of bioactive compounds present in the methanolic extracts of root-derived from in vivo grown plants of Muntingia calabura L

S No

RT

Compound name

Formulae

MWt

Area

Biological activity

1

22.36

hexadecanoic acid

C16H32O2

256

1.15%

Anti-inflammatory and arthritis (Abubakar et al., 2016)

2

24.58

cholest-4-en-6-on-3-ol

C27H46O2

400

63.7%

Anti-obesity activity (Shaheed et al., 2019).

3

26.22

1,12-Dibromododecane

C12H24Br2

326

0.40%

No activity

4

26.66

17-octadecynoic acid

C18H32O2

280

0.66%

Hypocholesterolemic, Anti-inflammatory, Anti-acne, Anti-coronary (Revathi & Dhanaraj, 2019)

5

27.22

(2R)-2,5,8-Trimethyl-2-[(4R,8R)-4,8,12-trimethyltridecyl]-3,4-dihydro-2H-1-benzopyran-6-ol

C28H48O2

416

1.34%

Anti-spasmodic (Lanuzza et al., 2017)

6

28.2

5-(2-bromopropan-2-yl)-2-methylcyclohexan-1-ol

C16H32O2

256

0.94%

No activity

7

28.71

cholestan-3-one-4,4-dimethyl-(5α)

C29H50O

414

0.25%

Anti-cancer and anti-inflammatory (Shaheed et al., 2019)

8

29.41

9-octadecenoic acid(Z)-phenyl methyl ester

C25H40O2

372

0.48%

Anti-ulcer genic, anti-androgenic, anti-inflammatory, anti-cancer (Shelke & Bhot, 2019).

9

29.87

(9Z)-octadec-9-enoic acid (Or)Oleic acid

C18H34O2

282

0.48%

Anti-pyretic, anti-nociceptive, anti-inflammatory (Aati et al., 2019)

10

30.19

(2R)-2,5,7,8-Tetramethyl-2-[(4R,8R)-4,8,12-trimethyltridecyl]-3,4-dihydrochromen-6-ol (or) Vitamin E

C29H50O2

430

8.26%

Anti-cancer and anti-oxidant (Shahaby, Zayat, Fattah, & Hefny, 2019)

11

33.73

2H-1-benzopyran-6-ol,3,4-dihydro-2,8-dimethyl(-2-4,8,12-trimethyl tridecyl)

C27H46O2

402

0.34%

No activity

12

34.7

androst 1-en-3-one,4,4-dimethyl-(5α)

C21H32O

300

0.87%

Breast and cancer cell activity (Yue et al., 2020)

13

35.1

A'-neogammacer-22(29)-en-3-one

C30H48O

424

5.82%

No activity

14

35.4

4,4,6a,6b,8a,11,11,14b-Octamethyl-1,4,4a,5,6

C30H48O

424

6.78%

No activity

15

35.9

1,2,5,14,18,18-hexamethyl-8-(prop-1-en-2-yl)pentacyclo[11.8.0.0²,¹⁰.0⁵,⁹.0¹⁴,¹⁹]henicosan-17-ol (or) Lupeol

C30H50O

426

7.3%

Anti-prostate and anti-cancer (Yang et al., 2020).

The compound, 5-hydroxy-7-methoxy-2-phenyl-4H-1-benzopyran-4-one, possesses anti-cancer, anti-prostate and anti-inflammatory activities (Marques, Silva, Marques, & Braga, 2019; Wang et al., 2019). Similarly, a compound is known as (Naringenin) (2S)-5,7-dihydroxy-2-(4-hydroxyphenyl)-2,3-dihydro-4H-1- benzopyran-4-one is observed to have two biological properties like anti-tuberculosis and anti-cancer activities (Sahu et al., 2020). The major compound identified in the M. calabura leaf extract was found to be 2-{3-[(E)-2-(1H-indol-3-yl)ethenyl]-1,2,4-oxadiazol-5-yl}phenol 5.78% peak area, and have antioxidant and antitumor activity (Figure 1). These bioactive compounds are synthetic aromatic C-nucleoside derivatives developed by (Sadek et al., 2014). 2,2'-((1E,1'E)-(1,4- phenylenebis(azanylylidene))bis(methanylylidene))diphenol (Nitrilo methyl dyne) has been reported to have an anti-tumor activity (Kopoytkoba et al., 2019).The compound 7-hydroxy-3-methoxy-2-(4- methoxyphenyl)-4H-1-benzopyran-4-one that is identified to possess antioxidant activity from the bran of Thai black rice (Suttiarporn et al., 2016) and 5,7-dihydroxy-2-(4-hydroxyphenyl)-6-methoxy-4H-chromen-4-one reported to have strong antioxidant activity (Karker et al., 2019; Mu et al., 2009; Yausheva et al., 2019). 2,3,7,8-tetramethoxy-5,11-dihydro10H-dibenzo[a,d][7]annulen-10-one has anti-inflammatory activity (Fernandes et al., 2019). The compound, Anthraquinone, 1-(4-methylphenyl)anthracene-9,10-dione is identified to act against the cancer cell and has antibacterial activities (Chu et al., 2019). 8,9-dihydro-aflatoxin B1 has the mycotoxin activity (Aparna et al., 2012; Pok et al., 2020), and 1-Docosene has the anticancer activity of Xestospongia testudinaria. Some compounds have been identified, but their biological activity has not yet been discovered (Table 1).

Figure 1

GC-MS chromatogram of leaf methanolic extracts of Muntingia calabura and potent bioactive compound 2-{3-[(E)-2-(1H-indol-3-yl)ethenyl]-1,2,4-oxadiazol-5-yl}phenol (antioxidant and antitumor 5.78%).

https://www.nrfhh.com/f/fulltexts/145564/image1_min.jpg

GC-MS analysis of root methanolic extract

Of all the bioactive compounds exhibited, cholest-4-en-6-on-3-ol has the highest 63.7% retention time (24.58 min) and has anti-obesity activity (Shaheed et al., 2019). At the same time, another compound, Lupeol, was observed during prolonged retention (35.9 min), and the maximum area percentage was 7.36% (Figure 2). Similarly, a compound is known, 17-octadecynoic acid has phytochemical properties like Hypocholesterolaemia, anti-inflammatory, anti-acne, and anti-coronary activities from various plant extracts (Benzidia et al., 2019; Mishra & Patnaik, 2020; Revathi et al., 2019). (2R)-2,5,8-trimethyl-2-[(4R,8R)-4,8,12-trimethyltridecyl]-3,4-dihydro-2H-1-benzopyran-6-ol (β-Tocopherol) is one of the compounds that have anti-spasmodic activity (Lanuzza et al., 2017). The compounds like cholestan-3-one-4,4-dimethyl-(5α); 9-octadecenoic acid(Z)-phenyl methyl ester; (9Z)-octadec-9-enoic acid (Oleic acid); androst1-en-3-one,4,4-dimethyl-(5α) identified to possess anti-cancer, anti-inflammatory, antioxidant, antiulcer genic, antipyretic activities in various plant extracts of medicinally important plants (Safwat, Hamed, & Moatamed, 2018; Shelke et al., 2019; Yamuna, Abirami, Vijayashalini, & Sharmila, 2017; Yue et al., 2020).

Figure 2

GC-MS chromatogram of root methanolic extracts of Muntingia calabura, potent anti-obesity compound cholest-4-en-6-on-3-ol(63.7%) and anti-cancerous compound Lupeol (7.3%).

https://www.nrfhh.com/f/fulltexts/145564/image2_min.jpg

Cholest-4-en-6-on-3-ol and Lupeol are the two identified compounds with highest percent of compounds among all the compounds in root methanolic extracts of M. calabura and reported to possess anticancer, antioxidant, anti-prostate, anti-inflammatory activities in different plant extracts of medicinal plants (Geetha & Varalakshmi, 2001; Shahaby et al., 2019; Shaheed et al., 2019; Yang et al., 2020). There are a few identified compounds, 1,12-Dibromododecane; 5-(2-bromopropan-2-yl)-2-methylcyclohexan-1-ol; 2H-1-benzopyran-6-ol,3,4-dihydro-2,8-dimethyl(-2-4,8,12-trimethyl tridecyl); A'-neogammacer-22(29)-en-3-one; 4,4,6a,6b,8a,11,11,14b-octamethyl-1,4,4a,5,6 yet to be identified (Figure 2 and Table 2).

Figure 3

Effect of Muntingia calabura leaf methanolic extract on various bacterial cultures.

https://www.nrfhh.com/f/fulltexts/145564/image3_min.jpg

Figure 4

Effect of Muntingia calabura root methanolic extract on various bacterial cultures.

https://www.nrfhh.com/f/fulltexts/145564/image4_min.jpg

Antibacterial activity for leaf and root methanolic extracts

The antibacterial activity of leaf methanolic extracts (45µg/mL, 60 µg/mL, 75 µg/mL) and the control Streptomycin (10 µg/mL) was evaluated. The maximum antibacterial activity was shown towards E. coli at 75µg/mL concentration, followed by B. sphericus, P. vulgaris and P. fluorescens. As the concentration of the plant extract increased, the inhibition zone was also found to increase. (Figure 4; Figure 3). The leaf methanolic extract of M. calabura was considered the most effective extract with high anti-bacterial activity (Zakaria et al., 2010). In M. calabura, the ethanolic leaf extract has shown strong antibacterial activity against E. coli (Gurning et al., 2021). The root extracts of Carica papaya have shown the highest antibacterial activity against Salmonella typhi among tested gram-negative and gram-positive bacterial strains (Doughari, Elmahmood, & Manzara, 2007).

CONCLUSION

This study revealed the presence of various bioactive molecules in the leaf and root methanolic extracts of M. calabura. Leaf methanolic extracts yielded thirty-eight compounds, while root extracts yielded a total of fifteen compounds based on their molecular weight, retention time, and peak area. The leaves of M. calabura contain the highest concentration of the antioxidant and antitumor compound is 2-{3-[(E)-2-(1H-indol-3-yl)ethenyl]-1,2,4-oxadiazol-5- yl}phenol, followed by roots of cholest-4-en-6-on-3-ol and Lupeol. These bioactive substances have been shown to have antibacterial effects against gram-positive and gram-negative bacteria. According to this study, the abundance of phytochemicals and bioactive substances in M. calabura makes it a potential source of medicines.

Conflicts of interest

Conflict of interest the authors declare that there are no conflicts of interest in this study.

Author contributions

SV conducted the experimental work, writing the article, PC analyzed the data and designed the manuscript helped in experimental work, and SV designing Chromatogram figures and interpretation the data. ST extended overall guidance and finalized the manuscript.