INTRODUCTION:

The developing resistance of microbial infections to traditional antibiotics has prompted the investigation of alternative therapeutic agents, especially Zingiber officinale Roscoe, or ginger, which has attracted significant attention for its numerous medical characteristics.

Historically utilized throughout several cultures, ginger demonstrates potential antibacterial properties that may offer useful remedies for combating diseases. The bioactive components in ginger, including gingerol and shogaol, contribute to its medicinal properties, which several studies have established for their efficacy against inflammation and bacterial infections¹. Previous studies demonstrate that extraction procedures utilizing ethanol provide elevated quantities of these phytochemicals, hence augmenting antibacterial efficacy². This article will examine Zingiber officinale's extraction and phytochemical screening procedures, assessing its antibacterial activity to support its use as an effective alternative for treating microbial diseases. Zingiber officinale Ginger, also referred to as roscoe, has a long history in traditional medicine and is valued for its many medicinal benefits. Its use extends back to thousands of years, especially in Asian cultures where it was used as a staple cure for respiratory and digestive disorders in addition to being used as a spice. Ginger's ability to adapt in herbal medicinal methods is demonstrated by the fact that it has been used historically in a variety of forms, including fresh, dried, and powdered. Various phytochemical studies have shown that ginger contains substances including shogaol and gingerol, which have strong antibacterial properties and support the plant's therapeutic benefits³. The gingerols were shown to be the main active ingredients in the fresh ginger rhizome. Ginger is detected by the senses due to two different chemical groups: volatile oils and non-volatile pungent chemicals. Sesquiterpene hydrocarbons make up the majority of ginger's volatile oil components, with zingeberene accounting for 35%, curcumene for 18%, and farnesene for 10%⁴.

Gingerol Shogaol

Zingiberene Paradol

Fig 1. components of Zingiber officinale (Ginger).

The unique flavor and scent of ginger are attributed to a number of these volatile oil components [Fig:1]. Gingerols, shogaols, paradols, and zingerone are examples of non-volatile pungent chemicals that give the tongue a "hot" feeling.

The main active ingredients in the fresh rhizome were found to be the gingerols, a group of chemical homologs distinguished by the length of their unbranched alkyl chains. Furthermore, the main strong ingredients in dried ginger are the dehydrated version of the gingerols and another homologous series called shogaols. Similar to gingerol, paradol is created when shogoal is hydrogenated. Oleoresins are an additional component. Vitamins, minerals, lipids, and waxes are all present in ginger⁵.

Table 1: Taxonomy, Distribution, and Botanical Description⁶

Kingdom:	Plantae
Division:	Magnoliophyta
Class:	Liliopsida
Order:	Zingiberales
Family:	Zingiberaceae
Genus:	Zingiber
Species:	Zingiber officinale Roscoe

Taxonomy is a practice and science concerned with classification or categorization which describe in Table 1. Active ingredients including phenolic and terpene chemicals are prevalent in ginger. The primary phenolic chemicals found in ginger are paradols, shogaols, and gingerols. Gingerols, including 6-, 8-, and 10-gingerol, are the main polyphenols found in fresh ginger. Gingerols can become comparable shogaols with heat treatment or prolonged storage. After being hydrogenated, shogaols can change into paradols⁷. Ginger contains several other phenolic compounds, including 6-dehydrogingerdione, zingerone, quercetin, and gingerenone-A⁸.

Fig 2: A fresh ginger (Zingiber officinale)

The public has been seriously threatened by the spread of infectious illnesses caused by bacteria, fungi, and viruses as a result of antibiotic resistance. To combat several harmful germs, a number of herbs and spices have been transformed into natural, potent antimicrobial compounds. Recent reports have indicated that ginger possesses antiviral, antifungal, and antibacterial properties⁹. Ginger essential oil contains chemicals with lipophilic qualities that increase the permeability of the cytoplasmic membrane and cell wall while also causing fungus to lose membrane integrity. Furthermore, Aspergillus flavus growth, aflatoxin formation, and ergosterol synthesis were all effectively inhibited by ginger essential oil¹⁰. By decreasing ergosterol production and altering membrane integrity,

ginger essential oil successfully suppressed Fusarium verticillioides growth in in-vitro research. Additionally, it may reduce the synthesis of fumonisin B1 and fumonisin B2¹¹. Ginger has therefore been shown to prevent the growth of several viruses, fungi, and bacteria. These effects may be primarily associated with the inhibition of viral attachment and internalization, ergosterol production, and bacterial biofilm formation (Table 2).

MATERIALS AND METHODS:

Plant and other materials:

Collect a fresh ginger about 1kg of Zingiber officinale Roscoe species from a local market. Assemble a chemicals and glassware that are Ethanol (Denatured), Chloroform, Petroleum ether and iodine flask, beaker, conical flask, measuring cylinder, sieve, butter paper, filter paper.

Fig 3: Chemical stock

Methods:

a) Processing (Drying) of collected Z. officinale rhizomes:

The rhizomes were properly cleaned two or three times using fresh water. Using a small cutter mill that was run by hand, ginger was chopped into tiny bits. Currently, the tiny ginger pieces were air-shaded and allowed to dry for 12hours without being exposed to sunlight, followed by a 72hour night cycle at each location. Following that, the partially dried ginger was allowed to air dry for seven more days at room temperature of 25±2°C and 45±5% relative humidity in a clean, cold, dry, and dark environment¹⁶. Periodically, the ginger was weighed using an electronic digital scale to gauge weight loss, and the moisture loss was calculated. When the ginger was thoroughly dried, it was noted that no fungus should form on the garlic or ginger. For future research, the dried ginger was kept in a dark, airtight container¹⁷.

Fig 4: Dry ginger (Zingiber officinale)

b) Size reduction of Z. officinale rhizomes:

A hammer mill with built-in sieves was used to break up the dried plant material (rhizomes). Between 30 and 40 meshes, they were reduced to an extremely coarse powder particle size. The extremely coarse powder form was kept at a low, regulated temperature and relative humidity in an airtight, covered glass container out of the light¹⁸.

(a) (b)

(c) (d)

Fig 5: (a, b) Powder form by using hammer mill, (c, d) coarse powder by using sieves

c) Extraction Techniques:

In maceration, the basic extraction technique, the 50gm plant-prepared raw material is soaked in a 70% solvent [50gm of raw material dip into 250ml of solvent on behalf of ratio 1:5(w/v)] of interest in a coarse or powdered state for at least three days at room temperature with periodic stirring after which it was filtered. The different solvent was used separately i.e. Petroleum ether, Chloroform, Water. After collecting and evaporating the macerate at 60°C using a rotary vacuum evaporator, it was concentrated on a water-bath at 60–70°C to produce a thick extract^19,20,21.

Fig 6: Extraction technique by maceration

PHYTOCHEMICAL SCREENING OF EXTRACT:

Using the method described test for alkaloid, saponin, Tannin, Phlobotannin, flavonoid, cardiac glycoside, steroid and Terpenoid, were carried out.

1. Flavonoid Test:

The samples' aqueous filtrate was mixed with five milliliters (5ml) of diluted ammonia solution, and then concentrated H₂SO₄ was added. An observation of yellow coloring was interpreted as proof that flavonoids were present²².

Fig 7: Flavonoid test

2. Alkaloid Test :

Mayer's test: A few drops of Mayer's reagent were added to 1 ml of filtrate. When alkaloids are present, a yellow-colored precipitate will form²².

Fig 8: Alkaloid test

3. Phenolic Test:

Ferric chloride test: The extracts were treated with 3 to 4 drops of ferric chloride solution. Formation of bluish black colour indicates the presence of Phenols²³.

Fig 9: Phenolic test

4. Terpenoids Test:

2 ml of chloroform was added to 0.5 g of the extract, and then 3 ml of concentrated H₂SO₄ was carefully added to create a layer. The presence of terpenoids was suggested by the interface's reddish-brown colouring²³.

Fig 10: Terpenoids Test

5. Tannis Test:

100ml of distilled water was used to mix 5g of extract and 5ml of honey before they were filtered. The filtrate was treated with ferric chloride reagent. Tannin was present when a blue-black or blue-green precipitate formed²³.

Fig 11: Tannis test

6. Steroids Test:

0.5g of extract was mixed with 2milliliters (2ml) of acetic anhydride, and 2ml of sulfuric acid was added via the test tube's sides. The color of the mixture was then checked for a shift from violet to blue-green²³.

Fig 12: Steroids test

7. Glycosides Test:

5g of each of the extracts and 5ml of honey was dissolved in 2ml glacial acetic acid containing a drop of ferric chloride solution. This was underplayed with 1ml concentrated H2SO4. A brown ring of the interface indicates a deoxy-sugar characteristic of cardenolides. A violet ring may appear below the brown ring, while in the acetic acid layer, a green ring may form just gradually spread throughout this layer²⁴.

Fig 13: Glycosides test

PREPRATION OF CLUTURE MEDIA AND ISOLATION:

Nutrient agar, which was produced and sterilized at 121 °C and 14 pounds for 14-30 minutes, was the antibacterial medium. Prepare nutrient agar media (meat extract, peptone, sodium chloride, distilled water, agar) and maintain pH level by 7.2±0.229. Pre-sterilized petriplates with a 90 mm diameter were filled with 20–24ml of autoclaved nutritional agar medium under aseptic conditions, and the plates were left to harden in the presence of UV light. Pre-incubation is then applied for 24hours to the prepared plates²⁵.

Fig 14. Agar media plate placed in Biological Oxygen Demand incubator (BOD)

Isolation of Escherichia coli from Urine:

To reduce contamination, a urine sample was taken using the clean-catch midstream procedure in a sterile container. A calibrated inoculating loop was then used to inoculate the material onto a sterile Nutrient Agar plate. In order to isolate individual colonies, the streaking technique was used. The plate was incubated for 24 hours at 37°C following inoculation^26,27. Bacterial growth was seen on the nutrient agar plate after incubation. Round, wet, and off-white or pale colonies were the hallmarks of colonies thought to be Escherichia coli. Gram staining, which showed Gram-negative short rods, a feature of E. coli, further validated these colonies. Particularly in situations of suspected urinary tract infections, this technique aids in the first detection and isolation of E. coli from urine^28,29.

(a) View at 10x without stain (b) View at 100x with stain

Fig 14: microscopy images of Escherichia coli.

ANTI-BACTERIAL STUDY AND DETERMINIATION OF ACTIVITY:

The extract was made in three distinct dilutions of 50 mg/ml, 100mg/ml, and 150mg/ml. These dilutions were tested for antibacterial activity in conjunction with 30 mg/ml of the standard antibiotic (ciprofloxacin) that was dissolved in distilled water³⁰. Antimicrobial activity was determined using the first Agar well diffusion technique, which is in accordance with the guidelines suggested by the Clinical and Laboratory Standards Institute (CLSI). They took the pre-incubated plates and used a spreader to disseminate the germs through them³¹. A metallic hollow road was used to bore the well into agar plates once the bacteria had dried. After that, 30µl of the common antibiotic ciprofloxacin and 100µl of extracts were placed onto sterile discs and incubated for 24 hours. The widths of the inhibitory zones on each plate were measured in millimetres following incubation³².

Measurements of zone of inhibition:

The antibacterial spectrum of the extracts was evaluated in terms of the zones of inhibition surrounding the wells following plate incubation. Antibiotic-induced zone of inhibition widths was contrasted with those generated by the plant extract. During the tests, the average zone diameter was measured. Cache plate findings were measured by measuring the zone diameter (mm) generated by plant extract of various solvents after 24-26 hours of incubation at 37°C³³

Fig 15: Agar plate

RESULT AND DISCUSSION:

Phytochemical screening:

The phytochemical screening test was conducted using different solvents such as ethanol, chloroform, petroleum ether and crude extract of Zingiber officinale Roscoe rhizome were summarized in Table 3. The result obtained from this study pointed that the tannins, alkaloids, Flavonoids, phenol, terpenoids, steroids, glycosides.

Table 3. Phytochemical screening of Zingiber officinale.

S. No	Test	Ethanol	Chloroform	Pet. ether
1.	Flavonoids	+	-	-
2.	Alkaloids	+	+	-
3.	Phenol	+	+	-
4.	Terpenoids	+	+	+
5.	Tannins	+	-	-
6.	Steroids	-	+	+
7.	Glycosides	+	-	-

Indicate: (+) present, (-) negative

Antibacterial:

The antibacterial activity of crude extract of Zingiber officinale Roscoe rhizome was tested by agar well diffusion method. The extract of plant leaves has been found to be potent against Escherichia coli. The antibacterial activity of Zingiber officinale Roscoe rhizome extract of different solvent is summarised in Table 4. Ethanol extract shown the maximum inhibition zone against Escherichia coli. (22mm) in 150mg/ml concentration and minimum against petroleum ether extract (16mm). As in 100mg/ml maximum inhibition in ethanol extract (18mm) and chloroform extract (15mm) and petroleum ether (12mm). Then inhibition in petroleum ether extract has not shown well proper anti-bacterial activity.

Table 4: Minimum inhibitory concentration of Zingiber officinale Roscoe plant

Extract	Standard (Ciprofloxacine) in mm	50 mg/ml	100 mg/ml	150 mg/ml	MIC
Ethanol	30mm	12mm	18mm	22mm	100 mg/ml
Chloroform	30mm	10mm	15mm	19mm	150 mg/ml
Pet. Ether	30mm	8mm	12mm	16mm	>150 mg/ml

Graph 1. Zone of inhibition vs. Concentration

CONCLUSION:

Phytochemicals isolated from Zingiber officinale Roscoe rhizome leaf extract may have antibacterial properties, according to the study mentioned above. The extract of certain solvents may include antimicrobial substances. The plant's antibacterial properties can be investigated further for potential application in the management of bacterial infections. It is possible to utilize the raw extract of Zingiber officinale Roscoe rhizome to combat certain bacteria. Zingiber officinale Roscoe rhizome crude extract may be a superior substitute for traditional antibacterial food additives. It has also long been established that Zingiber officinale Roscoe rhizome leaf crude extract possesses antimicrobial properties.

CONFLICT OF INTEREST:

The authors declare that there is no conflict of interest regarding the publication of this research work. The study was conducted independently without any financial or commercial support that could influence the results. All findings and interpretations are based solely on the experimental data, ensuring objectivity and integrity in the presentation of the research outcomes.

DISCLAIMER STATEMENT:

There are no conflicting interests, according to the authors. Products that are often utilized in research were employed in this study. The writers and product manufacturers do not have any conflicts of interest.

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Received on 23.05.2025 Revised on 11.09.2025

Accepted on 12.11.2025 Published on 22.01.2026

Available online from January 29, 2026

Asian J. Pharm. Res. 2026; 16(1):44-50.

DOI: 10.52711/2231-5691.2026.00006

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

Constituent	Study Type	Potential Mechanisms	Ref
Ginger essential oil	In vitro	Reducing ergosterol biosynthesis; affecting membrane integrity; decreasing the production of fumonisin B1 and fumonisin B2	11
Ginger essential oil	In vitro	Reducing ergosterol biosynthesis; affecting membrane integrity; inhibiting the production of aflatoxin	10
Gingerenone-A and shogaol	In vitro	Inhibiting the activity of 6-hydroxymethyl-7, 8-dihydropterin pyrophosphokinase	12
Gingerenone-A and shogaol	In vitro	Affecting membrane integrity; inhibiting biofilm formation	13
Ginger extract	In vitro	Inhibiting biofilm formation, glucan synthesis, and adherence	14
Ginger extract	In vitro	Blocking viral attachment and internalization	15