Phytochemical Investigation and Pharmacological Studies on The Stem of Jasminum Grandiflorum

Jasminum grandiflorum (Spanish jasmine or Royal jasmine) is a medicinally significant plant widely used in traditional and Ayurvedic medicine. While its flowers and leaves are well known for their therapeutic benefits, the stem also contains bioactive compounds that contribute to its pharmacological properties, including antipyretic and antifungal activities. The stem of J. grandiflorum is believed to possess fever-reducing properties due to the presence of flavonoids, alkaloids, and phenolic compounds. These bioactive molecules help regulate body temperature by inhibiting the release of pyrogens and reducing inflammation. Traditional healers have used extracts from the plant to manage fevers associated with infections and inflammatory conditions. The antifungal potential of J. grandiflorum stem is attributed to phytochemicals such as tannins, saponins, and essential oils. These compounds exhibit inhibitory effects against fungal pathogens by disrupting cell membranes and interfering with fungal growth. The stem extracts have been explored for their ability to combat fungal infections affecting the skin and other body systems. The medicinal significance of Jasminum grandiflorum stem highlights its potential as a natural alternative for treating fever and fungal infections. Further scientific research is necessary to validate its efficacy and explore its mechanisms of action.

METHODOLOGY

Physico Chemical Evaluation ^[1-4]

The moisture content, total ash, water soluble ash, acid insoluble ash, alcohol soluble extractive value, water soluble extractive value and loss on drying were determined as part of its physiochemical parameters. 

Preliminary phytochemical screening of extract

This is a standard set of qualitative phytochemical tests used to identify the presence of alkaloids, glycosides, flavonoids, and phenolic acids, terpnoids, saponins in ethanolic extract of stem of Jasminum grandiflorum. These tests on specific chemical reaction that produce characteristic color changes or precipitates, indicating the presence of different classes of bioactive compounds.

TLC or Thin Layer Chromatography-

System Components

TLC system components consist of

1. TLC plates, preferably ready-made with a stationary phase: These are stable and chemically inert plates, where a thin layer of stationary phase is applied on its whole surface layer. The stationary phase on the plates is of uniform thickness and is in fine particle size.
2. TLC chamber. This is used for the development of the TLC plate.
3. Mobile phase. This comprises of a solvent or solvent mixture (Toluene: Ethyl acetate: Formic acid) [5:4:1]
4. Detecting agent: Iodine chamber, UV light

7.5 High Performance Liquid Chromatography (HPLC) ^[51]

The HPLC Methods are follows:

Make: Waters HPLC System,Vienna,Austria

Detector: Waters 2545 Quaternary Gradient Module with 2998 Photodiode Array Detector (waters G.M.B.H., Veinna, Austria).

Column: Sunfire (4.58x250mm)C18 column.

Software: The data were collected and processed with the software Empower 2.0.

Sample preparation:1mg/ml concentration of the compound was taken and dissolved with HPLC-grade methanol.Compounds were then filtered through a syringe filter ( PVDF Filter membrane with polypropylene Housing pore size 0.2µm,whatman,GE Healthcare) for HPLC analysis.

Sample volume: 20µl

Moblie phase: Methanol: Water (50:50)

Flow rate:1ml/min.

• Invitro Antifungal Activity

1. Agar Well Diffusion Method

a. Potato Dextrose Agar Medium

The potato dextrose agar medium was prepared by dissolving 20 gm of potato influsion, 2 gm of dextrose and 1.5 gm of agar in 100ml of distilled water. The dissolved medium was autoclaved at 15 lbs pressure at 121°C for 15 minutes. The autoclaved medium was mixed well and poured onto 100mm petri plates (25-30 ml/plate) while still molten.

PROCEDURE

Petri plates containing 20ml potato dextrose agar medium was seeded with 72 hr 000culture of fungal strain (Aspergillus aculeatus, Cryptococcus neoformans) with different concentration of sample SJ (500, 250, 100 and 50 μg/ml) was added. The plates were then incubated at 28°C for 72 hours. The anti-fungal activity was assayed by measuring the diameter of the inhibition zone formed around the wells. Amphotericin B was used as a positive control. The values were calculated using Graph Pad Prism 6.0 software (USA).

• Invitro Antipyretic Activity

Nitric Oxide Radical Scavenging Assay ^[7,8]

Procedure

The extracts were prepared from a 50 mg/mL (SJ) crude extract. These were then serially diluted with DMSO to make concentrations from 500−10 μg/mL of the samples. These were stored at 4°C for later use. The 150 µl extract was mixed with an equal volume of freshly prepared Griess reagent (150µl). Control samples without the extracts but with an equal volume of buffer were prepared in a similar manner as was done for the test samples. After 30 minutes of incubation period, 100 μL of the reaction mixture was transferred to a 96-well plate. The absorbance was measured at 540 nm using a UV-Vis microplate reader (Molecular Devices, GA, USA).

Formula

Nitic Oxide Scavenge (%) = A control - A test/A control x 100

             A control   -   Absorbance of control

             A Test        -   Absorbance of test sample

RESULTS AND DISCUSSION

Physico-Chemical Evaluation

Physicochemical parameters are mainly used in judging the purity and quality of the powdered drug. The ash values of a drug gave an idea of the earthy matter or the inorganic composition and other impurities present along with the drug. The extractive values are primarily useful for the determination of exhausted or adulterated drug. The Physico-chemical standards of stem of Jasminum grandiflorum are listed below.                                                  

Data showing the Physico-chemical standards of Jasminum grandiflorum

Percentage Yield of Total Extract                                                             

Percentage yield, colour and consistency of ethanolic extract of Jasminum grandiflorum

Chromatographic Evaluation

Thin layer chromatography (TLC)

Figure 10.1: Thin Layer Chromotography

Figure 10.2: High Performance Liguid Chromatography

Table 10.4: Data showing the RT values of Jasminum grandiflorum stem extract

• Antifungal Activity

Agar well diffusion method

Fig:10.3 Effect of sample SJ against A.aculeatus

Fig:10.4 Effect of sample SJ against C.neoformans.

Table:10.5 Means of zone of inhibition obtained by sample SJ against A.aculeatus,C.neoformans

ANTIPYRETIC ACTIVITY

sample: Nitric Oxide Radical Scavenging Assay

A. OD Value at 540 nm

Figure 10.5: OD Value at 540nm

Figure 10.6: Percentage of inhibition

Table 10.8: C. IC50 Value of tested sample:46.16µg/

Figure 10.7: IC50 Value of tested sample:46.16µg/ml