Extraction of Hibiscus rosa sinensis Flower

Shubham Porte¹, Vinayak Kaushik¹, Geetanjali Sahu², Sharang Bali²

¹M. Pharma, School of Pharmacy, Chouksey Engineering College,

Lalkhadan, Bilaspur - 495004 Chhattisgarh.

²Assistant Professor, School of Pharmacy, Chouksey Engineering College,

Lalkhadan, Bilaspur - 495004 Chhattisgarh.

*Corresponding Author E-mail: shubham.sp.porte@gmail.com, iamvinayak49@gmail.com, geetanjali.sahu574@gmail.com

ABSTRACT:

Hibiscus rosa sinensis a widely known ornamental and medicinal plant has the history of traditional medicinal uses throughout the world, and is still used in many ways due to diverse medicinal properties, this plant has also been used as a showy flower.  Flavonoids, Alkaloids, Anthracyanins Tannins Saponins and other phytochemicals have shown significant therapeutic effects such as antioxidants, anti-inflammatory, antimicrobial and wound healing. This experiment is focuses on the extraction of these phytochemical compounds and preliminary phytochemical screening (Identification test) from Hibiscus rosa sinensis flower with the ethanol as the solvent running through Soxhlet apparatus, dried flower powder was subjected to continuous hot percolation method up to 48hours at the control temperature in between 70-80°C. The extraction method was proven to be highly effective in extracting the thermo stable phytochemicals, the final extract was evaporated and dried, test was performed such as Shinoda test, ferric chloride test and Wagner’s test to confirm the presence of certain phytochemical groups. These findings solidify the therapeutic properties and potential for future studies in the Hibiscus rosa sinensis. This experiment contributes to the understanding of the plants extraction process, chemical profile, medicinal properties and possible use advance drug delivery systems as the Phytopharmaceutical product.

KEYWORDS: Hibiscus rosa-sinesis, Soxhlet Apparatus, Phytochemicals, Wound-Healing, Alkaloids, Flavonoids Shinoda test, Ferric chloride test.

1. INTRODUCTION:

Hibiscus rosa sinensis is commonly found plant in the India, used for many proposes including medicine, ornament and offering, the vibrant color of this flower makes it more prone to gardening plant as well. Aside from the many other properties this has significantly medicinal properties. The leaves are used in Indian medicine for emollient, aperitif, anodyne, laxative and leaf decoction and stem bark are used for abortion system.¹ The medicinal importance of plants in the treatment of human ailments is immense and has been so since the dawn of civilization.² According to the World Health Organization, a medicinal plant is nothing but any plant which, in one or more than one of its organs, contains substances that can be used for therapeutic purposes, or which are major precursors for chemo-pharmaceutical and herbal pharmaceutics semi synthesis.³ This plant is considered as Mesophyte leads to its availability in every part of the country, it is native to China grown widely in India and found in different color for their showy flowers.^4,5 HRS is a tropical plant that grows best in warm, humid climates with average temperatures of 26°C and annual rainfall of 900mm.⁶ Extraction of herbal drugs is the process of separating the desired bioactive compounds from plant material, such as leaves, flowers, roots, seeds using various techniques and solvent.⁷ Therapeutic properties of this plant is widely distributed, it is used as the medicinal plant not only India but all around the world which includes Abortion, Antifertility, Diuretic, Constipation, Hair Oil, Shampoo, Antidiabetic, Anti-inflammatory, Analgesic, Antimicrobial, Diarrhea, Antipyretic, Wound Healing Cosmetics, it also contain bioactive compounds capable of reducing inflammation and oxidative stress, making it a potential adjunctive therapy for COPD. By targeting the molecular mechanisms of inflammation through natural compounds, it is possible to mitigate the symptoms and progression of COPD, providing a complementary approach to conventional              treatments. ^{8,9,10,11,12,13} Plant-derived compounds have played and will continue to play avital role in the process of drug discovery, particularly in the development of novel analgesics. However, evaluating the pharmacological effects of herbal crude extracts remains a viable research technique for the discovery of novel medications.¹⁴ The dried root of Hibiscus syriacus L. are used as a fork medicine in the orient for the cure of hematochezia, dysentery, obstruction due to wind phlegm, and vomiting of food.¹⁵ The ability of extract to absorb UV radiation and hence proved its UV protection ability. The extract showed a prominent absorbance at 200-250nm, while good absorbance at range of 260-320nm and moderate absorbance was reported at range of 300-380nm.¹⁶ Hibiscus rosa-sinesis L is a small plant belonging to family Malvaceae, flowers are red coloured, large, cyclic hypogynous, pedicilate, complete, hermaphrodite, actinomorphic, peutamerous.¹⁷

1.1 Common Names:⁴

Hindi	Gurhal, Mandaar, China Rose, Devifool, Jasun, Jasat, Arahul
English	Hibiscus, Chinese Rose, Shoe Flower Plant
Sanskrit	Japa, Java, Rudrapuspa, Aundrapuspa, Trisandhya
Marathi	Jasvand, Jasud
Malayalam	Ayamparathi, Chemparathi
Telgu	Java pushpamu, Dasara, Dasanamu, Mandara
Tamil	Sembaruth, Sapattuu
Bengali	Jaba, Jiwa, Oru, Panchmukhi
Gujrati	Jasua, Jasunt, Jasud
Kannad	Dasvala, Dasaala
Punjabi	Jasum, Jaipushpa, Gurhal

1.2 Taxonomical Classification¹⁸

Table 1. Taxonomical classification of Hibiscus rosa sinensis plant.

Kingdom	Plantae-Plants
Subkingdom	Tracheobionta-Vascular plants
Super division	Spermatophyta-Seed plants
Division	Magnoliophyta-Flowering plants
Class	Magnoliopsida-Dicotyledons
Subclass	Dilleniidae
Order	Malvales
Family	Malvaceae-Mallow family
Genus	Hibiscus L.-Rosemallow
Species	Hibiscus rosa sinensis

1.3 Distribution:

Origin of this Plant found to be Tropical Asia or more specifically historically it is believed to be originated from China, the natural distribution of the parent species is seemed to be over 4000km, this showed how much extensive human-mediated movement and breeding of plants.

Asia: This plant is widely found in the gardens across India, most specifically in the southwestern regions and also in the countries like Sri Lanka (tropical area), Thailand, Philippines, Myanmar, China, and Pakistan.^17,19 Historically it is cultivated Chinese provinces such as Fujian, Guangdong, Guangxi, Hainan, Sichuan, Taiwan and Yunnan and the Japan.^20,21

Africa:

These are mostly due to its showy flower and distributed alongside of south Africa.^9,10

Pacific Islands:

This plant is highly valued by Polynesian communities and cultivated in these regions for long period of time.

Europe:

Commonly it is grown in the Med Terrance region and mostly popular as showy flower or houseplant in the northern European areas, and cultivated mainly in the cold winters.

1.4 Nutrients Composition:

Table 2. Nutrient Composition of Hibiscus rosa sinensis Flower (per 100g fresh weight).²²

Nutrient	Amount (per 100g)
Moisture	83.00%
Protein	1.54 g
Fat	0.35 g
Crude Fibre	1.50 g
Carbohydrate	13.71 g
Β-carotene	54.02 µg
Vitamin C	7.502 mg
Calcium	4.32 mg
Potassium	236.45 mg
Iron	1.48 mg
Zinc	0.82 mg

1.5 Phytochemical Composition:

Table 3. Phytochemical composition in the Hibiscus rosa-sinensis plant.

Phytochemical Class	Specific Compound(s)	Weight / Concentration	Source
Anthocyanins	Cyanidin-3-sophoroside	213.64mg/100g (red)	Yuan, Yet al.^23,24
	Cyanidin-3,5-diglucoside	877.04mg/100g (fresh petals)
	Cyanidin-3-sophoroside-5-glucoside	37.29mg/100g (blue (vacuum dried)
Flavonoids	Rutin – 4104µg/g	Total: 61.59mg QE/g extract	Rajendran et al^25
	Kaempferol – 361.9µg/g
	Myricetin – 50.7µg/g
	Quercetin – 7.6µg/g
	Kaempferol-3-glucoside – 5.1mg/g
	Quercetin-3-glucoside – 2.0mg/g
	Catechin – 1.5mg/g
	Epicatechin – 1.1mg/g
	Apigenin-hexuronide – 0.3mg/g
Phenolics	Total Phenolic Content 35.82mg GAE/g extract	3146mg/100g^26	Dávila et al^26
		4598.16mg/100g^26	Barde et al^27
Tannins	Ellagic acid, Gallotannins	0.073 mg/g	Zulkurnain et al^28
Alkaloids	Hibiscine	~0.51%	Zulkurnain et al^29
Organic Acids	Citric acid – up to 991.4mg/g DW, Malic, Tartaric, Oxalic acids	Varies	Carcea, M^.29
Saponins	Saponin glycosides	Present (abundant)	Zulkurnain et al^28
Glycosides	Flavonoid and cardiac glycosides	Present	Sharma, Y et al^30
Terpenoids	Taraxeryl acetate	Present	Zulkurnain et al^28
Phytosterols	β-Sitosterol, Campesterol, Stigmasterol	Present	Zulkurnain et al^28

1.6 Therapeutic Effects:

Table 4. Therapeutic properties and their mechanism of action of Hibiscus rosa-sinensis plant.

Therapeutic Property	Plant Part Used	Mechanism of Action / Activity	Author / Source
Anti-inflammatory	Flower and Leaf	Reduce the production of pro-inflammatory mediators (COX, TNF-α, IL6), and inhibit inflammatory cell migration.	Geeganage and Gunathilaka et al^31
Antioxidant	Flower	Neutralizes free radicals via flavonoids, tannins, anthocyanins, phenolics and boosts antioxidants enzymes.	OS Falade et al^32
Anti-microbial	Flower, Leaf, Root	Damages Bacterial cell walls – disrupt microbial DNA/RNA synthesis.	Munir A et al^33
Anti-diabetic / Hypoglycemic	Leaf, Flower	Inhibits the carbohydrate metabolizing enzymes (α-amylase) – Improve insulin sensitivity.	Rani, Seema, et al^34
Antihypertensive	Flower, Leaf	Stimulate the enzyme Angiotensin Converting Enzyme-2 (ACE-2) and inhibit the calcium channel.	Amtghri et al^35
Antipyretic / Analgesic	Root, Flower	Inhibits prostaglandins synthesis – reduces fever and pain.	Aziz M.H.A. et al^36
Wound‑healing	Flower, Leaf	Promotes fibroblast proliferation, enhances collagen formation and induces skin tissue repair.	Shivananda N et al^37
Anticancer / Chemopreventive	Flower, Leaves	Triggers apoptosis in the cancer cells and inhibits tumor cell division as well as DNA replication.	Chang, Y. C., and Hsu, C. C et al^38
Antidepressant / CNS effects	Leaves, Flower	It is mediated through Dopaminergic, Adrenergic, Serotonergic mechanism Modulates CNS neurotransmitters - anxiolytic effect.	Shewale, Pallavi B et al^39
Antifertility / Abortifacient	Flower and Leaves	Modulates the estrogen receptors, inhibits implantation and ovulation in animal models.	Vasudeva, Neeru, and S K Sharma et al^40
Antifungal Activity	Flower, Root, Leaf	Inhibition of biofilm formation in the candida albicans and block ergosterol biosynthesis.	Mohana, Pallvi et al^41
Neurobehavioral (stress)	Flower	Modulation of neurotransmitters and Inhibition of MAO-A activity	Shewale, Pallavi B et al^39
Antidiarrheal	Leaf and Flower	Astringent action by slowing down intestinal motility and water loss.	Missoum, A et al^43
Anti-arthritic / Anti-gout	Leaf and Flower	Inhibits Inflammatory cytokines in the joints, Reduction of uric acid and oxidative stress.	Patel, M., and Sharma, A^44
Hepatoprotective	Flower (powder)	Reduce the liver fat (steatosis) and protect against liver damage.	Ujianti, Irena et al^45

2. MATERIALS AND METHODS:

2.1 Chemicals: Ethanol (95v/v%), Methanol, conc. HCL, Lead Acetate, Zinc Metal, Ferric Chloride, Wagner’s Reagent, NaOH.⁴⁶

2.2 Apparatus: Soxhlet extraction assembly, including a 5L round-bottom flask, extractor body, Allihn-type condenser, Heating mantle with a rheostat for temperature control, vacuum pump.^47,48

2.3 Plant Materials: Hibiscus rosa-sinensis flower are collected in local plant nursery identified by the Botanist. The Flower petal are washed with fresh water and dried in the sun light for two days further it was placed under yellow light for some hours for complete evaporation of any kind of moisture. The dried petals are grinded to make coarsely powder.⁴⁹

Picture 1 – Dried flower petals of Hibiscus rosa-sinensis plant

Picture 2 – Grinded Powder of dried flower petals of Hibiscus rosa-sinensis plant

2.4 Preparation of Plant Extract: The extraction of Hibiscus rosa sinensis was carried out using Soxhlet extraction method, dried hibiscus flower powder (500g) was taken. Solvent used was ethanol (95% analytical grade), the procedure was conducted using a standard laboratory-scale Soxhlet apparatus, which consisted of a 2 L capacity round-bottom flask, a Soxhlet extractor body of appropriate size to accommodate the sample load, an Allihn-type condenser to facilitate solvent reflux, and a rheostat-controlled heating mantle for precise temperature regulation.^50,51,52

Picture 3 and 4. Soxhlet Extraction Apparatus for Hibiscus rosa-sinensis

The detailed extraction procedure was as follows:^{53, 54}

1. A total of 500g of the dried H. rosa-sinensis flower powder was carefully loaded into a large-format cellulose extraction thimble. Care was taken to ensure the powder was not overly compacted, which could impede the free flow of solvent through the plant material and reduce extraction efficiency.

2. The loaded thimble was securely placed within the main chamber of the Soxhlet extractor.

3. One liter of 95% ethanol was measured and transferred into the 2 L round-bottom flask.

4. The apparatus was then fully assembled, with the flask connected to the bottom of the extractor and the condenser fitted to the top. A continuous flow of cold water was circulated through the outer jacket of the Allihn condenser.

5. The heating mantle was activated, and the ethanol was heated to a gentle boil (approximately 78°C). The extraction process was allowed to proceed continuously for an extended duration of 48hours.

This process operates on a principle of continuous solvent reflux: the ethanol vaporizes from the flask, travels up the side-arm of the extractor, condenses on the cold surface of the condenser, and drips down onto the plant material in the thimble. The chamber containing the thimble gradually fills with the hot solvent, which dissolves the soluble phytochemicals. Once the solvent level reaches the top of the siphon tube, the entire volume of solvent, now enriched with the extract, is siphoned back into the boiling flask. This cycle repeats continuously, ensuring that the plant material is repeatedly washed with fresh, pure, hot solvent, driving the extraction process towards completion.⁴²

2.5 Physical Characterization and Yield Calculation:

To find out the efficacy of the extraction process the percentage yield of the crude extract is calculated by its initial mass relative to plant material so the yield wascalculated by using this formula.⁵⁵

Yield (%) = (Final weight of crude extract / Initial weight of flower powder) × 100

Table 5. Identification test of Phytochemical compounds.

Test Name	Target Compound	Typical Chemicals Used	Used/Alternative Chemical	Observation
Anthocyanin Test^58	Anthocyanins (flavonoid pigment)	Acid/Base treatment (e.g., HCl/ NaOH), or pH differential test	Lead acetate (alternative method)	Red in acid, blue in base; yellow ppt = flavonoids
Shinoda Test^59	Flavonoids	Magnesium turnings + Conc. HCl	Zinc granules + HCl (alternative)	Red/pink coloration
Ferric Chloride Test^60	Phenolics/ Tannins	1% Ferric chloride solution	No alternative	Green, blue, or black coloration
Wagner’s Test^61	Alkaloids	Iodine + Potassium Iodide (Wagner’s reagent)	Prepared Wagner’s reagent	Reddish-brown precipitate
Froth Test^62	Saponins	Ethanol + Distilled water	No alternatives	Formation of Froth Layer

Picture 5 – Ethanolic Extract of Hibiscus rosa-sinensis flower

The obtained plant crude extract was found to be viscous, dark reddish-brown, oily, greasy, semi solid material.⁴⁴ Its organoleptic properties were exhibiting acidic and astringent aroma, with fruity undertone. The Appearance dark reddish-brown color suggests that a highly concentration of natural pigments, mostly phenolic compounds like anthocyanins, flavonoids and their derivatives, which are found high on the colored hibiscus flowers.⁵⁶

2.6 Phytochemical Screening: The following test are performed in the plant crude collected after extraction process to detect presence of various compounds.⁵⁷

2.7 Findings:

2.8.1 Extraction Yield and Observation:

The exhaustive 48-hour Soxhlet extraction of 500g of Hibiscus rosa-sinensis flower powder with 95% ethanol yielded 92.5g of the crude extract. This calculates to a percentage yield of 18.5% (w/w). This yield is substantial and reflects the efficiency of the prolonged, high-temperature extraction method in removing a significant portion of the soluble constituents from the plant matrix. A study of Sandra Pimentel-Moral et al. (J Pharm Biomed Anal. 2018)⁵² employing a conventional hydro-extraction (reflux) of hibiscus leaves reported a lower yield of 12.26%, while a more advanced microwave-assisted extraction yielded 15.16%. The higher yield observed in the present experiment is therefore consistent with the more aggressive and exhaustive nature of the 48-hour Soxhlet protocol.

2.8.2 Phytochemical Profile of Extract:

The qualitative phytochemical screening of the prepared crude ethanolic extract yielded positive results for all five major classes of secondary metabolites tested.

The specific observations from these tests were as follows:

Test for Alkaloids (Wagner's Test):

The addition of Wagner's reagent to the extract with conc. HCL solution resulted in the formation of a dense, reddish-brown precipitate, confirming the presence of alkaloids.

Test for Flavonoids (Shinoda Test):

The reaction of the extract with Zinc metals and HCl produced a reddish amber color, providing a positive indication for the presence of flavonoids.

Test for Phenols and Tannins (Ferric Chloride Test): With the addition of 1% ferric chloride solution to the extract result in dark blue-black color, confirming the presence of phenolic compounds, including tannins.

Test for Saponins (Froth Test):

Vigorous shaking of the diluted extract in water generated a stable and persistent layer of honeycomb-like froth that exceeded 1cm in height and did not dissipate after 10 minutes, indicating the presence of saponins.

Test for Anthocyanins (Acid-Base Test):

The extract demonstrated characteristic pH-dependent color changes, turning a light red/yellowish upon acidification with HCl, then after addition of NaOH solution it later turned yellowish green confirming the presence of anthocyanins.

DISCUSSION:

Hibiscus Rosa Sinensis is plant that are used widely in the world to cure many types of disease and problems mostly it is used for the showy flower but studies suggest it has the great therapeutic properties. In the present studies it is found to be the plant has abundance of flavonoids, Alkaloids and other Phytochemicals, the confirmation of these compounds validates for the traditional use of this plant for medicinal purposes. The presence of Flavonoids, Anthocyanins and phenolic acids gives use evidence of its antioxidant and anti-inflammatory activities, Saponins and Tannins contributes for the antimicrobial, antidiarrheal and wound healing properties, the detection of alkaloids is crucial, as this class of nitrogenous compounds is associated with a vast range of potent pharmacological effects, including the analgesic and neuroprotective properties that have been reported for H. rosa-sinensis extracts.

CONCLUSION:

This research effectively proved that a 48-hour uninterrupted hot extraction by a Soxhlet apparatus with 95% ethanol is a very effective process for yielding a high-yielding (18.5% w/w) crude extract from powdered flowers of Hibiscus rosa-sinensis. The following qualitative phytochemical analysis gave strong and conclusive proof that this ethanolic extract is a chemically complex mixture with plenty of diverse classes of secondary metabolites. The screening indicated the presence of alkaloids, flavonoids, phenols and tannins, saponins, and anthocyanins. The results are generally concordant with the documented phytochemical profile of H. rosa-sinensis reported throughout the scientific literature and thus confirm the validity of extraction and analytical procedures used. In addition, the outcomes point towards significant discrepancies with particular studies, highlighting the considerable influence that factors such as extraction method and plant cultivar can affect the final chemical makeup of a plant extract.

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Received on 30.07.2025      Revised on 27.08.2025

Accepted on 17.09.2025 Published on 08.10.2025

Available online from October 17, 2025

Asian J. Pharm. Tech. 2025; 15(4):339-345.

DOI: 10.52711/2231-5713.2025.00050

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