1. INTRODUCTION:

The Asian tree Ginkgo biloba also known as the maidenhair tree is living fossil known for its medicinal properties and otherbenefit to the human health.This tree is originated in the Permian Era about 250 million years ago and shown in the Chinese Materia Medica 5000 year ago.^1,2,3,4The ginkgo biloba tree is the surviving member of its family Ginkgoaceae.⁵

Ginkgo biloba wasuse as traditional Chinese medicine and ornament but now itshas emerged as a extensively researched medicinal plant with diverse therapeutic application.⁶Following the modern research it was found that ginkgo biloba has numerous bioactive compound responsible for its pharmacological effect but the main chemical constituent are flavonoids and terpene trilactones.^7,8 the flavonoid comprises the large fraction of bioactive compoundtypically 24% in which the major compounds are quercetin, kaempferol, and isorhamnetin which are present as glycosides.⁹The terpene trilactone mainly contains ginkgolide A, B, and C along with bilobalide.^6,10,11

Recent studies have concentrated on the potential of Ginkgo biloba in addressing neurodegenerative diseases, particularly Alzheimer’s Disease.¹²Ginkgo biloba extract holds the ability to ameliorate fluoride toxicity.¹³its extracts have been used pharmaceutically as anti-inflammatory, anti apoptic, and anti-aging.¹⁴GA is an effective antagonist of Platelet Activating Factor (PAF) and reduces nerve damage due to ischemia and excitotoxic injury.¹⁵Concurrent administration of Ginkgo biloba reduced the plasma malondialdehyde and increased the levels of antioxidant enzyme.¹⁶Chinese experimental studies on the effect of Ginkgo biloba extract on oral cariogenic bacteria found that it can inhibit Streptococcus mutans.¹⁷

The Soxhlet extraction method integrates the advantages of the percolation and reflux extraction, which utilizes the principle of siphoning and reflux to continuously extract the herb with fresh solvent.¹⁸ Soxhlet extraction is a broad extraction technique that is commonly applied to analytes that are sufficiently thermally stable.¹⁹

1.1. Synonyms⁷

Table 1: Sources

Worldwide (English)	Maidenhair tree
Japan	Ginkyo
France	“L’arbre aux Quaranteecus” and “Noyer Du Japon”

1.2. Taxonomy and Classification:

Table 2: Taxonomical classification of ginkgo biloba plant

Kingdom	Plantae
Subkingdom	Tracheobionta
Superdivision	spermatophyta
Division	Ginkgophyta
Class	Ginkgopsida
Order	Ginkgoales
Family	Ginkgoaceae
Genus	ginkgo
Species	Ginkgo biloba L.

1.3 Distribution and Cultivation:

Native habitat – China:

The natural distribution of ginkgo biloba is confined to china, where it persists in three distinct glacial refugia²⁰:

Specific wild population location:

· Zhejiang province: contain the primary wild population²¹

· Dalou Mountains (Southwestern China): fragments of original natural range in valley and lower mountain slopes²²

· Qianzhong Altiplano (middle Guizhou): 126 wild locations with various community types

Korea:

Korea had the first overseas introduction of ginkgo tree in tang dynasty in six centuries.²⁰

Japan:

Introduce by China multiple time over the last two millennia through human mediated dispersal and due to commercial connection between China and Japan in seventh century.²⁰

Europe:

Utrecht, in Netherlands, was the site of the first ginkgo tree planting in Europe in around 1730.After 1730, ginkgo biloba tree has been planted across Europe.^20,23

America: America’sfirst ginkgo tree was planted in 1784 at Hamilton’s private estate in Philadelphia. Then its planted massively in late 19^th and 20^th centuries.^20,23

1.4 Nutrient Content:

Table 3: nutrient composition of ginkgo biloba L.²⁴^,6

Nutrient		Amount (per 100)
Protein		12.27±0.24g
Fat		4.75±0.22g
Carbohydrates		72.98±0.20g
saturated fatty acid		0.319g
Vitamins	Vitamin c	15mg
	Vitamin B1	0.22mg
	Vitamin B2	0.09mg
	Vitamin B3	0.16mg
	Vitamin B6	0.328mg
Minerals	Manganese	0.113mg
	Calcium	2mg
	Copper	0.274mg
	Iron	1mg
	Zinc	0.34mg
	Magnesium	27mg
	Sodium	7mg
	Phosphorus	124m
Water		54.2g

1.6 Therapeutic Effect:

Table 5: Therapeutic effect and their mechanics of action of ginkgo biloba

Therapeutic effect	Mechanism of action	Active compound	Clinical evidence	references
Neuroprotection	* Free radical scavenging and antioxidant activity * Enhanced Bcl-2 expression * Inhibition of cytochrome C release * Suppression of caspase-3 levels * Reduction of 8-hydroxydeoxyguanosine	Flavonoids (quercetin, kaempferol) Terpene lactones (ginkgolide, bilobalide)	Alzheimer's disease models show improved cognitive function and reduced neurodegeneration	[7][29][30]
Cognitive Enhancement	* Increased acetylcholine, dopamine, and serotonin levels * Acetylcholinesterase inhibition * Improved cerebral blood flow * Enhanced neuroplasticity	EGb761 standardized extract Flavonoid glycosides Terpene lactones	Clinical trials show improvement in memory, attention, and early-stage dementia	[30][32][33]
Cardiovascular Protection	* Vasodilation and improved microcirculation * Antiplatelet aggregation * Endothelial-dependent and independent vasorelaxation * Myocardial protection	Ginkgolides A, B, C Bilobalide Flavonoids	Effective in peripheral vascular disease, diabetic complications, and metabolic syndrome	[34][35]
Anti-inflammatory activity	* Reduction of C-reactive protein (-1.5 mg/L) * Decreased IL-6 (-16.86 pg/mL) * Reduced TNF-α (-4.19 pg/mL) * Inhibition of inflammatory signaling pathways	Quercetin Kaempferol Isorhamnetin Ginkgolides	Meta-analysis confirms significant anti-inflammatory effects in clinical trials	[36][37]
Antioxidant Effects	* Scavenging of reactive oxygen species * Enhanced antioxidant enzyme activities * Lipid peroxidation inhibition * Cellular membrane protection	Flavonoids (primary) Proanthocyanidins Organic acids	Protective effects in ischemia-reperfusion injury and oxidative stress conditions	[38][39]
Platelet aggregation inhibition	* Platelet-activating factor (PAF) antagonism * Inhibition of thromboxane synthesis * Reduced platelet adhesion * Improved blood rheology	Ginkgolides (specific PAF antagonists) Bilobalide	Clinical applications in thrombotic disorders and vascular protection	[30][40][41]
Metabolic Effects	* Improved insulin sensitivity * Enhanced glucose metabolism * Lipid profile improvement * PPARG and DPP4 modulation	Flavonoid glycosides Terpene lactones Ginkgetin	Beneficial effects in metabolic syndrome and diabetes management	[42][43]
Ocular/Auditory Protection	* Improved ocular and cochlear circulation * Neuroprotection in sensory organs * Enhanced microvascular perfusion * Reduced oxidative damage	EGb761 Ginkgolides Bilobalide	Clinical applications in sudden hearing loss and retinal disorders	[34]
Pulmonary protection	* Inhibition of p38 MAPK pathway * Reduced pulmonary fibrosis * Anti-fibrotic effects * Improved lung function	Ginkgo biloba extract Active flavonoids	Systematic reviews show efficacy in idiopathic pulmonary fibrosis	[44][45]
Anti-aging Effect	* Cellular senescence inhibition * Mitochondrial protection * DNA damage prevention * Telomere protection	Ginkgetin Flavonoids Terpene lactones	Studies show cardioprotective effects and cellular anti-aging properties	[46][47]

2. MATERIAL AND METHOD:

2.1 Chemicals-Ethanol (95% v/v), chloroform, sulphuric acid, Methanol 50%, Magnesium chunk, conc. Hydrochloric acid, Ammonium solution, ferric chloride, distilled water

2.2 Apparatus-Round bottom flask, Soxhlet extractor body, extraction thimble, reflux condenser, heating mantle, retort stand with clamps and bossheads, rubber tubing, beaker, weighing balance filter paper, measuring cylinder, glass funnel, vacuum pump.⁴⁸

2.3 Plant Material-The dried Ginkgo biloba leaves were purchased from reputed online herbal store, Indian Jadibuti. A total of 500 g of ginkgo biloba leave were used in this experiment. plant material was visually inspected for quality then leaves were mechanically ground into coarse powder for the more efficient extraction. the powdered ginkgo biloba was stored in airtight container at room temperature in dry and dark environment until further use for extraction.

2.4 Preparation of Ethanolic aqueousextract of ginkgo biloba leave^49,50,51

Figure 1: Ginkgo biloba leave powder

Figure 2: Extaction from Ginkgo biloba leave powder

For the preparation of ginkgo biloba extract we use the Soxhlet extraction method. Following are the steps of extraction procedure:

1. Sample loading: an exact 500g of dried and powdered form of ginkgo biloba leave is weighed and transferred into large thimble. Since the fine powder has a chance to get into the siphon tube in extraction, the thimble’s top was the lightly plugged with glass wool. Then carefully insert the loaded thimble into Soxhlet extraction chamber.

2. Apparatus Assembly: The 2 Liter round-bottom flask was filled with 1liter of 95% of ethanol. ethanol is used as solvent in this extraction. The round bottom flask was fastened onto the Soxhlet extractor at the bottom and the condenser was attached to the upper opening of the extractor. To ensure continuous condensation of the solvent vapor, the inlet and outlet port of the condenser were connected to a cold-water supply. The whole setup was mounted on a clamped support retort stand and placed on heating mantle in the fume hood for the proper vaporisation of ethanol.

3. Extraction Process: the heating mantle was turned on and temperature was slowly increasing using the rheostat until ethanol in that flask boiled (~78%). Solvent vapour was rises through the side distillation tube into the condenser and condensed. The solvent evaporates through the thimble and after condensation drips into the powder plant material. The solvent level in the thimble chamber ascended, when the chamber fills to the siphon level, the extract returns to the flask. This cycle of extraction was continuously uninterrupted for 48 hours to ensure the extraction of targeted compound.

4. Post extraction: After the extraction period of 48 hour was over, switch off the heating mantle and leave it for cool down at room temperature. The spent plant material in the thimble was removed and discarded. The concentrated extract was collected in flask. Then the rotary evaporator saw use with 40-50ºC water bath. This gently remove the ethanol and the concentrated semi solid extract remain in flask.

3. Evaluation:

3.1. Yield Calculation of Extracted Material:

The Yield for Ginkgo Biloba extraction was calculated by using following equation⁴³:

Weight of extract from plant material

Yeild extract= ------------------------------------------- x 100

Weight of dried plant powder

% Yeild extract = (86/ 500 )x 100

% Yeild extract = 17.2%

3.2. Organoleptic and Physical characterisation:

· Consistency: At room temperature, the extract seen as sticky and highly viscous semisolid mass.

· Colour: The extract possessed a rich, dark brownish green colour.

· Odour: weak aromatic, earthly and due to flavonoids and terpenoids a slightly astringent smell

Figure 3: Extract of Ginkgo biloba leav

Figure 4: Identification test results

3.3. Phytochemical screening of extract:

Following identification tests are performed to detect the presence of targeted chemical compounds^52,53,54:

Test for Flavonoids (Shinoda test): 1.5ml of methanol add in 4ml of extract solution; a small magnesium chunk was added. Red colour was shown when add 5-6 drops of concentrated HCl were added that confirm the flavonoids are present.

Test for terpenoids: Add 0.2g of extract was add in 2ml of chloroform and mixed, then add 3ml of concentrated H₂SO₄. It was demonstrating the reddish-brown colour which confirm the terpenoids is present in extract.

Test for tannins (ferric chloride test): 0.5g of extract was mixed in 10ml of distilled water to make the dilute solution then added 3-4 drop of FeCl₃. Formation of a blue-black or green blackish colour in the presence of ferric chloride precipitate was taken as evidence for the presence of tannins.⁵⁵

Test for saponins: 0.5ml of filtrate mixed with 5ml of distilled water, shaken for 30sec.⁵⁶The froth was form which was the sign of the saponins presence.

4. RESULT AND DISCUSION:

The total of 132.5g of crude extract was get from the Soxhlet extraction of 500g of dried powder ginkgo biloba leave with 95% ethanol used as solvent for the duration of 48 hour. The corresponding yield percentage was 26.5%.

The extract appearance was dark brownish green highly viscous semisolid with slightly stringent smell. The stringent smell was due to presence of flavonoid and terpenoid.

In the phytochemical study of ginkgo biloba extract we detected the presence of various phytochemical constituents and the following are the test result for the targeted class of compounds.

Extract material/ tests	Shinoda test (flavonoids)	Test for terpenoids	Ferric chloride test (tannins)	Forth test (saponins)
Ginkgo biloba extract	+	+	+	+

+ = Present, - = Absence

5. CONCLUSION:

In this study, successfully demonstrated that the Soxhlet Extraction with the use of 95% ethanol as a solvent was so effective for the extraction of various phytochemical compounds out of ginkgo biloba leave. The 26.5% yield was shown effectiveness of this methodology and the extract that revealed the clear presence of various bioactive compound display that this method is recovering the phytochemical constituent with high efficiency from the plant material. And the recovery of polar and semi polar compound max due 95% ethanol. The method and duration of extraction was the major factor for the quality and quantity of the compounds present in the plant extract.

Through the phytochemical screening of the extracted material, disclose the various class of compound present in ginkgo biloba in which some major classes are flavonoids and terpenoid with the various other compound. Which are highly used for their pharmacological and therapeutic potential.

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Received on 28.08.2025 Revised on 23.10.2025

Accepted on 05.12.2025 Published on 04.07.2026

Available online from July 18, 2026

Asian J. Pharm. Tech. 2026; 16(3):228-234.

DOI: 10.52711/2231-5713.2026.00032

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

Phytochemical class	Compounds	Weigth/ concentration	Source
Flavonoids	Quercetin, kaempferol, isorhamnetin, glycosides	24%	[10], [25]
terpenoids	Ginkgolides A, B, C, J, M, (2.8 - 3.4%) Bilobalide (2.6 - 3.2%)	6%	[10][11]
proanthocyanidins	Catechin, Epicatechin, Procyanidin	2 -7%	[26]
carboxylic acid	quinin acid, chlorogenic acid, ascorbic acid, shikimic acid, gallic acid, protocatechuic acid, vanillic acid, isovanillic acid, ferulic acid	13%	[26], [27]
Ginkgolic acid	Alkylphenols	0.0005%	[28]
lignan		70mg/ml
polyprenol and polysaccharides		variable amount
inorganic constituents	mineral residue
water, solvent
unknown