INTRODUCTION:

The uses of plant bio-actives act as potent medicines for curing ailments, fighting infections and healing that predominates worldwide. The biologically active plant metabolites contain useful compounds having medicinal value that are utilized for treating various disorders affecting mankind. The botanical medicines due to their structural diversity and wide availability in nature are one of oldest disciplines found within healing practice for various ailments¹. Jasmine plant commonly known as Chameli is an important sub-shrub that contains essential oils that are found in the leaves and flowering tops (Figure 1)^2,3.

The wide array of medicinal properties is identified in the plant due to presence of various phytochemicals. The phytochemicals present in the J. pubescens have potent antioxidant, antimicrobial, anti-fungal activity against infections⁴.The leaves of plant contains various active metabolites like monoterpene hydrocarbons, sesquiterpene hydrocarbons, diterpenes, non-terpene derivatives n-Peicosane, n-Tricosane, α-Pinene, Benzaldehyde, (E,E)-2,4-Heptadienal, Linalool, Nonanal,4-Terpineol, Methyl salicylate, (Z)-Jasmone, β-Caryophyllene, (E)-Geranyl acetone, α-Humulene, trans-Nerolidol, (E,E)−α-Farnesene, Abietadiene and trace elements as p-Cymene, Eugenol⁵. The preliminary phytochemical investigation and wide distribution J. pubescens plant in nature makes it effective against throat infections, renal disorder, diabetes, abdominal discomfort, ulcers, dysentery and diarrhea and improving menstrual flow⁶.

Fig. 1: J. pubescens Plant Leaves and Flowers⁷.

MATERIAL AND METHODS:

Plant materials and Authentication:

The fresh leaves of the J. pubescens were identified, procured from the herbal garden and authenticated by Botanist Mr. Radhashyam Kharol R.G. Mada Vidyalaya, Mandsaur, M. P. The leaves were dried under shade, powdered and stored for further study.

Collection of Organism:

The selected pathogens E. coli (ATCC-10531), B. subtilis (ATCC-21479), Ps. aeruginosa (ATCC-25619), S. aureus (ATCC-65388) for antimicrobial study were collected from Department of Pharmacognosy, Institute of Pharmaceutical Research, GLA University, Mathura, U.P., India.

METHOD:

Preparation of Leaves Extract:

The shade dried leaves were powdered and packed into Soxhlet apparatus and extracted using continuous hot percolation method with petroleum ether (Pet. E), methanol (MEOH) and water (Aq.) separately. The extraction process was continued for 72 h and crude extracts (Aq.-E, MEOH-E, Pet. E) obtained after extraction were stored for further investigation (Figure 2)^8,9.

Fig. 2: Various Extracts of Jasmine Leaves: Aq.-E [A], MEOH-E [B], Pet. E-E [C].

QUALITATIVE PHYTOCHEMICAL ANALYSIS:

The various phytoconstituents are extracted in leaves of J. pubescens and estimated for potential antimicrobial and antioxidant activity according^10-16.

QUANTITATIVE ESTIMATION OF ANTIOXIDANT ACTIVITY OF LEAF EXTRACTS:

DPPH method:

The antioxidant activity of Aq.-E, MEOH-E, Pet. E-E were determined using DPPH (2, 2-diphenyl-1-picrylhydrazyl) estimation method using standard ascorbic acid. The samples were quantitively analyzed by UV-VIS spectrophotometer (Shimadzu UV-1700, Japan) at 517nm for obtaining absorbance of standardized and other extracts^17,18.

Total phenolic content:

Aq.-E, MEOH-E, Pet. E-E were analyzed using standard gallic acid for determination of total phenolic content and determined spectrophotometrically^19,20.

Flavonoids:

The leaves of J. pubescens contain flavonoids due to antioxidant potential. The standard calibration curve was constructed using quercetin as a standard for determination of flavonoid content in various prepared extracts^21,22.

Ferric reducing ability of plasma (FRAP) method:

This method was employed for determination of antioxidant activity using gallic acid as a standard with various leaf extracts of J. pubescens^19,21.

ESTIMATION OF ANTIMICROBIAL ACTIVITY:

The antimicrobial activity of leaf extracts was determined by agar well diffusion method. The incubated bacterial cultures of microorganisms were mixed with molten agar media and poured in pre-sterilized agar plates. After solidification of prepared agar plates, sterile bores of 6mm diameters were made, suitably sealed with molten agar to prevent loss of extracts and filled with 0.45µl of 200mg/ml concentration of extracts. The ofloxacin is used as standard. Stock cultures were prepared using nutrient broth media and incubated for 24 h with loop full of cultures in BOD incubator. After overnight incubation, sterile cotton was used for swabbing over the agar (nutrient agar medium) plates to make uniform culture lawns. The plates were investigated after 2-3days for presence of any zone of inhibition (ZOI). The diameters of ZOI were measured using a hand-held digital caliper.

MIC (minimum inhibitory concentration) values of the methanolic extracts of J. pubescens were analyzed by nephlo turbidimetry method. The two-fold serial dilutions were made with methanolic extract having concentrations of 50, 25, 12.5, 6.25, 3.12 and 1.56 mg/ml, kept in 6 test tubes and suitably dissolved in DMSO (dimethyl sulphoxide). The nutrient broth media in each test tube containing methanolic extracts were incubated for 24 h at 37⁰C^23-26.

RESULTS AND DISCUSSIONS:

Phytochemical analysis:

The results of qualitative phytochemical analysis revealed that bioactive compounds were present in leaf extracts of J. pubescens (Table 1). The MEOH-E of leaves contains higher concentrations of tannins, steroids and quinones, Aq.-E contain higher conc. of alkaloids, betacynines, steroids and Pet. E-E contain glycoside respectively. The MEOH-E of leaves contains therapeutically active phytoconstituents and selected for determination of antioxidant and antimicrobial potential.

Table 1: Phytochemical Test of Secondary Phytometabolites.

Phytometabolite	Aq.-E^*	Pet. E-E^*	MEOH-E^*
Coumarins	+ve	-ve	-ve
Flavonoids	+ve	-ve	+ve
Tannins	+ve	-ve	++ve
Glycosides	-ve	-ve	+ve
Steroids	+ve	-ve	++ve
Terpenoids	-ve	+ve	+ve
Betacyanins	++ve	-ve	-ve
Cardiac glycosides	-ve	+ve	+ve
Saponins	++ve	-ve	+ve
Alkaloids	++ve	-ve	+ve
Phenols	+ve	-ve	+ve
Quinones	-ve	-ve	++ve
Carbohydrate	++ve	-ve	-ve
Protein and Amino acids	+ve	-ve	-ve

-ve: Absent; +ve: Present; ++ve: High concentration

*Aq.-E (Aqueous extract), MEOH-E (Methanolic extract), Pet. E-E (Petroleum ether extract)

Estimation of Antioxidant activity:

The antioxidant concentrations of MEOH-E were determined using DPPH method and found to be 12-35 mg/g respectively indicating free radical scavenging activity of Jamine leaves (Table 2).

Table 2: Antioxidant concentration of extract in different amount from DPPH assay

Extract	Amount (mg)	Antioxidant concentration of powder (mg/g)
MEOH-E of Leaves	10	12
	20	32
	30	35

Flavonoid, Total phenolic and antioxidant concentration of methanolic extract (FRAP) method:

The MEOH-E of Jasmine leaves contain flavonoids, total phenolic content and antioxidant concentrations (FRAP) as 31, 4 and 34 mg/g respectively. The results of study revealed that MEOH-E of J. pubescens have good antioxidant potential (Table 3).

Table 3: Antioxidant concentration of extract in different amount from DPPH assay.

Extract	Amount (mg)	Antioxidant concentration of powder (mg/g)
MEOH-E of Leaves	10	12
	20	32
	30	35

Antimicrobial activity of leaves extracts:

The antimicrobial activity test is an essential technique used in pharmacology to investigate the efficacy and potency of antimicrobial agents from herbal extracts against microorganisms. The results (Table 4) of antimicrobial activity depicted that MEOH-E showed significant antimicrobial activity as compare to other leaves extracts of J. pubescens. All extracts showed antimicrobial activity but MEOH-E showed highest antimicrobial action (expressed as ZOI) against S. aureus (25.7±0.84mm), as compare to other extracts. While the antimicrobial activity of Aq.-E was reported as S. aureus (19.3±0.65mm), B. subtilis (17.7±0.23mm), E. coli (14.3±0.46mm), P. aeruginosa (12.4±0.77mm) and Pet. E-E as S. aureus (13.7±0.33mm), B. subtilis (11.5±0.33mm), E. coli (9.1±0.86) and P. aeruginosa (8.8±0.29mm) respectively. The antimicrobial activity of standard drug was more as compare to J. pubescens (leaf) extracts.

The MEOH-E showed highest MIC value (3.12mg/ml) as compared to other extracts (Figure 3). Further, MEOH-E indicated significant antimicrobial activity against S. aureus because of presence of secondary phytometabolites like tannins, steroids, cardiac glycosides, phenols, flavonoids, alkaloids, terpenoids, saponin that can easily partition and interacts with bacterial cell wall, disrupt electron transport chain and results in cell lysis^23,27.

Table 4: Antimicrobial Activity of various Leaves Extracts of J. pubescens.

Test organism	Diameter of ZOI^# (mm)
Test organism	Aq.-E^*	MEOH-E^*	Pet. E-E^*	OFC^*
B. subtilis	17.7±0.23	22.8±0.65	11.5±0.33	30.7±0.12
E. coli	14.3±0.46	20.9±0.51	9.1±0.86	29.4±0.10
S. aureus	19.3±0.65	25.7±0.84	13.7±0.33	33.6±0.47
P. aeruginosa	12.4±0.77	18.1±0.45	8.8±0.29	28.3±0.61

^#N=3±S.D. (Standard deviation); ^*Aq.-E - Aqueous extract, MEOH-E - Methanol extract, Pet. E-E - Petroleum ether extract, OFC- Ofloxacin (as a Standard), ZOI-Zone of inhibition

Fig. 3: MIC of Methanolic Extract of J. pubscens Leaves.

CONCLUSION:

The present study for Jasmine leaves was successfully conducted and confirmed the presence of various secondary phytometabolites in Aq-E, MEOH-E, Pet. E-E using qualitative estimation. The leaves contain phenol, flavonoids and have showed good antioxidant potential, determined using FRAP and DPPH method. The methanolic extracts showed significant antimicrobial activity against S. aureus as compared to the other extracts of leaves. The potent antioxidant and antimicrobial activity of J. pubescens leaves make it a successful candidate for screening of newer herbal remedies for curing ailments and helping mankind.

ACKNOWLEDGEMENT:

The authors are highly appreciable to the management of GLA University Mathura, U.P. for providing assistance for carrying out the research work.

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

REFERENCES:

1. Colvard MD, Cordell GA. Rationalizing the Study of Plants for the Treatment of Oral Pain. Current Chemical Biology. 2008; 2(2): 140-152.

2. Sabharwal S, Sudan S, Ranjan V. Jasminum sambac Linn (Motia): A Review. International Journal of Pharmaceutical Research and Bio-Science. 2013; 2(5): 108-130.

3. Singh S, Jangde R, Daharwal SJ. Jasmine an Updated Review on Herbal Drug in Wound Healing. Research Journal of Pharmacy and Technology. 2019; 12(6): 3089-3097.

4. Thiruvengadam S, Nivedha S, Pujita V, Romauld SI. Detection of Antioxidant and Antimicrobial Activity of Leaf Extract of Jasminum azoricum. Research Journal of Pharmacy and Technology. 2018; 11(8): 3629-3632.

5. Temraz A, Cioni PL, Flamini G, Braca A. Chemical Composition of the Essential Oil from Jasminum pubescens Leaves and Flowers. Natural Product Communications. 2009; 4(12): 1729-1732.

6. Sabharwal S, Aggarwal S, Vats M, Sardana S. Jasminum sambac (Linn) Ait: Preliminary Phytochemical Screening and Wound Healing Investigation using Total Ethanol Stems Extract. International Journal of Pharmaceutical Sciences Review and Research. 2012; 17(1): 44-47.

7. http://mmmnaturals.blogspot.com/2016/07/types-of-jasmine_29.html (accessed on March 20, 2020)

8. Gupta R, Gupta J. Investigation of Antimicrobial Activity of Euphorbia Hirta Leaves. International Journal of Life science and Pharma Research. 2019; 9(3): 32-37.

9. Hemasudha TS, Yuvarani S, Rajakumari K. Combination of Azadirachta indica and Phyllanthus acidus - A complete Evaluation and Characterization Study. Research Journal of Pharmacy and Technology. 2019; 12(6): 3048-3052.

10. Britto JA, Gracelin DHS. Efficacy of Fruits of Jasminum grandiflorum Linn. Against Plant and Animal Pathogens. Asian Journal of Pharmaceutical and Clinical research. 2011; 4: 74-75.

11. Gupta R, Gupta MK, Bhandari A, Gupta J. Preliminary Pharmacognostical and Physicochemical Analysis: A Poly Herbomineral Formulation. International Journal of Drug Development and Research. 2014; 6(3): 85–92.

12. Thilagavathi T, Kathiravan G. Phytochemical Analysis and Antimicrobial Activity of Ethonolic Leaf Extract of Ficus racemosa Linn. Research Journal of Pharmacy and Technology. 2017; 10(2): 537-540.

13. Ruskin S, Ajina RS. Qualitative Phytochemical Screening and In-vitro Anthelmintic Activity of Adhatoda vasica (Acanthaceae). Research Journal of Pharmacy and Technology. 2017; 10(2): 414-420.

14. Nair DU, Saraswathy MP, Kishore N, Pully NR. Phytochemical and Antimicrobial Evaluation of Luffa cylindrica Linn. Leaf and Flower Extracts – An In-Vitro Study. Research Journal of Pharmacy and Technology. 2010; 3(2): 438-441.

15. Rahate KP, Padma R, Parvathy NG, Rengith V. Quantitative Estimation of Tannins, Phenols and Antioxidant Activity of Methanolic Extract of Imperata cylindrica. International Journal of Research in Pharmaceutical Sciences. 2013; 4(1): 73-77.

16. Gupta R, Gupta MK, Bhandari A, Gupta J. Preparation and Standardization of Polyherbomineral Formulation. International Journal of Drug Development and Research. 2014; 6(2): 211–9.

17. Prabha SB, Rao M, Kumar MRR. Evaluation of in vitro Antioxidant, Antibacterial and Anticancer Activities of Leaf Extracts of Cleome rutidosperma. Research Journal of Pharmacy and Technology. 2017; 10(8): 2492-2496.

18. Umavandhana R, Jayanthi S. Analysis of Phytochemical compounds and DPPH Radical Scavenging Activity of Dictyota dichotoma and Halimeda macroloba. Research Journal of Pharmacy and Technology. 2018; 11(8): 3440-3444.

19. Prakkash MAJ, Ragunathan R, Johney J. Evaluation of Bioactive Compounds from Jasminum polyanthum and its Medicinal Properties. Journal of Drug Delivery and Therapeutics. 2019; 9(2): 303-310.

20. Tiwari P, Patel RK. Estimation of Total Phenolics and Flavonoids and Antioxidant Potential of Ashwagandharishta Prepared by Traditional and Modern Methods. Asian Journal of Pharmaceutical Analysis. 2013; 3(4): 147-152.

21. Amini MH, Kalsi V, Kaur B, Khatik GL, Lobo R, Singh G, Agarhari UC, Yele S, Suttee A. Phytochemical Screening and Antioxidant Activity of Heracleum afghanicum Kitamura Leaves. Research Journal of Pharmacy and Technology. 2017; 10(10): 3498-3502.

22. Tiwari P. Phenolics and Flavonoids and Antioxidant Potential of Balarishta Prepared by Traditional and Modern Methods. Asian Journal of Pharmaceutical Analysis. 2014; 4(1): 05-10

23. Gupta J, Gupta R, Ahuja A. Formulation, Characterization and Antimicrobial Activity of Polyherbal Hand Wash. International Journal of Pharmaceutical Sciences and Research. 2020; 3: 45-47.

24. Kumar S, Navneet, Gautam SS. Screening of Antimicrobial Properties of Jasminum sambac Linn. Leaf Extracts Against Dental Pathogens. Research Journal of Phytochemistry. 2015; 9(4): 195-200.

25. Hemalatha M, Arirudran B, Thenmozhi A, Rao USM. Antimicrobial Effect of Separate Extract of Acetone, Ethyl Acetate, Methanol and Aqueous from Leaf of Milkweed (Calotropis gigantea L.). Asian Journal of Pharmaceutical Research. 2011; 1(4): 102-107

26. Shivsharan U, Ravva S. Antimicrobial Activity of Pomegranate Juice. Research Journal of Pharmacy and Technology. 2018; 10(11): 1-3.

27. Saraswathi K, Sivaraj C, Jenifer A, Dhivya M, Arumugam P. Antioxidant, Antibacterial Activities, GCMS and FTIR Analysis of Ethanol Bark Extract of Capparis sepiaria L. Research Journal of Pharmacy and Technology. 2020; 13(5): 2144-2150.

Received on 06.06.2020 Modified on 12.07.2020

Research J. Pharm. and Tech. 2020; 13(12):6073-6076.

DOI: 10.5958/0974-360X.2020.01058.6