1. INTRODUCTION:

1.1 Background:

Because herbal teas offer anti-inflammatory, immune-stimulating, and antioxidant properties, humans have been drinking them since ancient times. Combining different herbs can produce a range of bioactive compounds, such as polyphenols, flavonoids, alkaloids, and essential oils, which increase their medicinal potential and have synergistic effects¹. Plant parts that are high in bioactive substances including polyphenols, flavonoids, and terpenoids, such as leaves, flowers, roots, and bark, are used to make herbal teas. These substances have been shown to have antibacterial, anti-inflammatory, and antioxidant qualities that can aid in the management and prevention of a number of illnesses. For generations, herbal teas have been a mainstay of traditional medicine, providing a natural and comprehensive means of fostering general health and well-being. The creation of polyherbal tea formulations has drawn a lot of attention lately due to the rise in demand for plant-based products and the growing interest in alternative medicine. In addition to enhancing flavor, these herb combos offer practical advantages like better digestion, detoxification, and immunological support². Due to its distinct place in the cultures of many nations as well as its appealing flavor and aroma, tea is today the most popular beverage in the world. Growing consumer knowledge of the health advantages of tea intake has resulted in a resurgence of interest in the beverage³. In order to achieve a particular goal, such as relaxation, rejuvenation, or alleviation from a certain condition, among other things, the majority of teas may contain one or more herbal ingredients or a combination of herbal compounds⁴.

1.2 GUAVA LEAF (Psidium guajava):

Guava synonym is Guava it consists of fresh and dried leaves of Psidium guajava belonging to family Myrtaceae.

Fig.1. Guava Leaves (Psidium guajava)

Flavonoids (quercetin, kaempferol), tannins, terpenoids (eugenol, caryophyllene), and phenolic acids (gallic acid, ellagic acid) are among the bioactive substances found in guava leaves (Psidium guajava) that have anti-inflammatory, anti-bacterial, antidiabetic, and antioxidant qualities. These components assist the immune system and digestive health, among other medical advantages of guava leaves^5,6. The well-known tropical tree Psidium guajava, also known as guava, is widely farmed for its fruit. It has around 3,800 species and roughly 133 genera. All portions of Psidium guajava have long been used medicinally. In English, the plant is commonly referred to as "guava." Antimicrobial and antibacterial substances are abundant in it⁷. Because of their many health advantages, guava leaves—which come from the Psidium guajava tree—have long been utilized in traditional medicine. The leaves are a great option for herbal teas that support overall health because they include substances like flavonoids, antioxidants, and tannins. Particularly, guava leaves are well-known for their antioxidant, anti-inflammatory, and antibacterial qualities. They are commonly found in tea formulations that promote digestive processes, encourage heart health, and improve blood sugar balance. Furthermore, guava leaves are frequently mixed with other herbs like Tulsi, ginger, or lemongrass to produce a synergistic effect that optimizes the beverage's health advantages⁸. The tea's flavor, potency, and general therapeutic value are all improved by this multi-herb blend, which makes it a popular option for people who are health-conscious⁹. Guava leaf powder is a useful ingredient in formulations intended for diabetic patients because of its well-researched capacity to control blood sugar levels and enhance insulin sensitivity. Guava leaf extracts have been demonstrated in studies to inhibit the alpha-glucosidase enzyme, delaying the conversion of carbs to glucose and so lowering blood sugar increases following meals¹⁰.

1.3 LEMONGRASS (Cymbopogon citratus):

Lemongrass synonym is Indian verbena it consists of fresh and dried leaves of Cymbopogon citratus belonging to family Poaceae (Grass family).

Fig.2. Lemongrass (Cymbopogon citratus)

The antibacterial, anti-inflammatory, and antioxidant qualities of lemongrass (Cymbopogon citratus) are attributed to its essential oils, which are abundant in substances like citral (a blend of geranial and neral), limonene, and myrcene. Its therapeutic properties in culinary and medical applications are further enhanced by the presence of flavonoids, phenolic chemicals, and terpenoids. Lemongrass is used extensively for its antibacterial, anti-inflammatory, and antioxidant qualities because of its high citral concentration. Citral works very well to promote relaxation and lessen inflammation. Lemongrass has antibacterial qualities, but it also includes flavonoids like luteolin that aid in detoxifying and improve digestion. Because of its purifying qualities, adding lemongrass powder to a polyherbal blend can enhance digestion overall, lower stress levels, and support healthier skin¹¹. In many herbal blends, lemongrass powder is used to boost digestion, lower inflammation, and strengthen the immune system. It is a useful ingredient in formulations meant to prevent infections and enhance general wellness because of its inherent antibacterial and antifungal qualities¹². Lemongrass's phenolic chemicals, which aid in the body's defense against free radicals, are responsible for its antioxidant action. This defense mechanism promotes general cell health, delays the aging process, and may help avoid oxidative stress-related chronic illnesses¹³. Citral and luteolin are two of the chemicals that give lemongrass powder its anti-inflammatory qualities. Lemongrass is a beneficial addition to formulations aimed at ailments like arthritis, muscle soreness, or other inflammatory diseases because of these components that help reduce inflammation¹⁴. Stress Reduction and Relaxation: Lemongrass helps lower anxiety and enhance the quality of sleep because of its relaxing and soothing effects on the nervous system. Lemongrass is a beneficial component to polyherbal compositions intended to reduce stress, encourage relaxation, and improve mental clarity because of these qualities¹⁵.

1.4 TULSI (Ocimum sanctum):

Tulsi synonym is Holy basil it consists of fresh and dried leaves of Ocimum sanctum Linn. belonging to family Lamiaceae.

Fig.3.Tulsi (Ocimum sanctum)

Essential substances like ursolic acid, eugenol, and flavonoids like apigenin are found in tulsi. These substances support the immune-stimulating, adaptogenic, anti-inflammatory, and antioxidant qualities of tulsi. Tulsi powder improves respiratory health, increases immunity generally, and aids in stress reduction. Because of its adaptogenic qualities, holy basil, also known as tulsi, is highly valued in traditional medicine. It strengthens the immune system and improves the body's reaction to stress. Tulsi's anti-inflammatory, antioxidant, and antibacterial properties are attributed to its flavonoids, including apigenin, ursolic acid, and eugenol. It also helps lower stress levels and has respiratory advantages. The addition of tulsi powder to a polyherbal blend promotes immunity, mental clarity, and general well-being¹⁶. Because of its high concentration of phenolic components, including as flavonoids and eugenol, tulsi is mainly recognized for its antioxidant qualities. These substances aid in the body's defense against dangerous free radicals, avoiding oxidative stress, which is connected to aging and chronic illnesses. Tulsi is also well-known for its antibacterial and anti-inflammatory properties, which make it a useful component of polyherbal preparations that promote skin, respiratory, and infection prevention. Additionally, tulsi aids in detoxifying and digestion, which increases its usefulness in holistic herbal remedies¹⁷. Tulsi is well known for its adaptogenic qualities, which enable the body to handle stress and preserve equilibrium under trying conditions. Tulsi leaf powder has been shown in studies to enhance cognitive performance, foster mental clarity, and lessen anxiety brought on by stress. By regulating cortisol levels, the stress hormone, it improves the body's resistance to emotional, mental, and physical stress. Because of this, it is a beneficial herb in polyherbal preparations intended to lessen fatigue, anxiety, and depression¹⁸.

1.5 GINGER (Zingiber officinale):

Ginger synonym is Sunthi it obtained from rhizome of Zingiber officinale belonging to family Zingiberaceae.

Fig.4.Ginger (Zingiber officinale)

Bioactive substances found in ginger, such as gingerol, shogaol, and paradol, are well-known for their strong anti-inflammatory, anti-nausea, and antioxidant qualities. Because of these ingredients, ginger powder helps to improve circulation, reduce inflammation, ease nausea, and support digestive health¹⁹. Because it improves digestion and reduces nausea, ginger powder is essential for digestive health. By boosting the synthesis of digestive fluids, the chemicals gingerol and shogaol stimulate the digestive tract and aid in the breakdown of food and nutritional absorption. Ginger has also been shown to improve stomach motility, which lessens the discomfort brought on by bloating and indigestion. Ginger powder enhances other herbs with digestive qualities when added to polyherbal formulations, resulting in a synergistic combination that supports the best possible digestive function²⁰. Because ginger contains phenolic components like gingerol and shogaol, it has strong antioxidant qualities. These substances aid in the neutralization of free radicals, which are oxidative stress-producing, unstable molecules that lead to aging and a number of chronic illnesses, including as cancer and heart disease. Ginger promotes tissue protection and cellular health by lowering oxidative damage, which is essential for preventing illnesses associated with oxidative stress²¹. Ginger's anti-inflammatory qualities are among its best-established benefits. It has been demonstrated that the active ingredient gingerol dramatically lowers inflammatory indicators, which helps afflicted people feel better and have better joint function²².

2. MATERIAL AND EQUIPMENTS:

2.1 Plant Materials:

Table No.1. Plant Materials

Sr No	Plant Material	Procured From
1	Guava Leaf Powder (Psidium guajava)	Home Garden
2	Lemongrass Powder (Cymbopogon citratus)	Home Garden
3	Tulsi Powder (Ocimum sanctum)	Home Garden
4	Ginger Powder (Zingiber officinale)	Local Market

Guava Leaf (Psidium guajava) used as Antioxidant Properties. Lemongrass (Cymbopogon citratus) used as Stress Relief and Relaxation. Tulsi (Ocimum sanctum) used as boosts immune function or immune support. Ginger (Zingiber officinale) used as promotes optimal digestive function.

2.2 Equipment and Other Materials:

Table No.2. Equipment and other Materials

Sr No	Equipment and Other Material
1	Grinding Machine
2	Weighing Balance
3	Hot Air Oven
4	Tea Bags
5	Sieve (No.40 and 20)
6	UV Spectrophotometer

3. METHODOLOGY:

3.1 Review on Key Ingredients:

1) Guava Leaf (Psidium guajava): Quercetin, kaempferol, gallic acid, and other polyphenolic chemicals are abundant in guava leaves and contribute to their strong antioxidant properties. Extracts from guava leaves have been shown in studies to increase the activity of antioxidant enzymes, decrease lipid peroxidation, and scavenge free radicals. For example, studies show that the polysaccharides in guava leaves help prevent oxidative stress by lowering cell mortality and preventing the production of reactive oxygen species²³.

2) Lemongrass (Cymbopogon citratus): Citronella, geraniol, and myrcene are among the chemicals found in lemongrass that have long been recognized for their relaxing properties. A study that was published in the "Journal of Clinical Psychopharmacology" discovered that aromatherapy using lemongrass oil considerably decreased participants' anxiety levels. Lemongrass tea has also been used for its gentle sedative qualities, which help people relax and fall asleep²⁴.

3) Tulsi (Ocimum sanctum): Holy basil, another name for tulsi, is well known for its adaptogenic qualities. The "Journal of Ayurveda and Integrative Medicine" published a systematic review that showed that tulsi improves immunological response by raising white blood cell counts and regulating cytokine production. Additionally, preclinical research has demonstrated that tulsi can prolong mouse life periods and prevent ulcers brought on by stress, suggesting its function in stress adaptation²⁵.

4) Ginger (Zingiber officinale): Bioactive substances like gingerol, shogaol, and zingerone, which have potent anti-inflammatory and antioxidant qualities, are commonly found in ginger. "Frontiers in Nutrition" released a critical review that examined how these substances can alter the pathways of inflammation and oxidative stress, potentially lowering the risk of chronic illnesses²⁶.

3.2 Collection of Raw Materials:

Fresh ginger rhizomes (Zingiber officinale), tulsi leaves (Ocimum sanctum), lemongrass leaves (Cymbopogon citratus), and guava leaves (Psidium guajava) were collected from local markets and herbal gardens. All materials were selected based on their freshness, cleanliness, and absence of physical damage or microbial contamination.

3.3 Preparation of Herbs Powder:

All herbs were thoroughly washed under running water to remove dust and impurities, followed by rinsing with distilled water. Dried in a shade-drying room at 40°C for 2-3 days. After drying, each herb was ground into a powder using a mechanical grinder. Each powder was sieved through a 40-mesh sieve to obtain uniform particle size. The powders were sifted to ensure uniform particle size and stored in airtight containers.

3.4 Formulation of Polyherbal Tea:

Various formulations were made by mixing the powdered herbs in different proportions. The proportions were varied to achieve the desired taste and therapeutic properties. The Poly herbal tea fills into sachets.

Table No.3.Formulation Table

Sr. No.	Ingredient	F1	F2	F3
1	Guava Leaf Powder	3.5gm	2gm	3gm
2	Lemongrass Powder	2.5gm	3.5gm	3gm
3	Tulsi Powder	2gm	2.5gm	2gm
4	Ginger Powder	2gm	2.5gm	2gm

3.5 Preparation of Poly Herbal Tea:

All herbal ingredients (Guava leaf powder, Lemongrass powder, Tulsi powder and Ginger powder) were weighed accurately. Mix the powders thoroughly using a blender or manually in a clean, dry container to ensure uniformity. Pack the blended powder in tea bag sachets. Fill tea bags (1.5–2g per bag), seal. Store in a cool, dry place away from sunlight.

Fig.5.Poly herbal tea Sachets

4. RESULT AND DISCUSSION:

Table No.4.Observation Table

Sr No	Parameters	Observation
1	Color	Slightly yellowish brown
2	Odor	Aromatic
3	Taste	Slightly Spicy, Refreshing
4	pH	6
5	Bulk Density	0.42 g/ml
6	Tapped Density	0.50 g/ml
7	Carr’s Index	16 %
8	Hausner Ratio	1.19
9	Angle of Repose	32.61
10	Ash Value	57.5 %
11	Water Insoluble Ash	2.24%
12	Acid Soluble Ash	1.5%
13	Diffusion Time with 1to 2 times dipping	1-2 Min.

1) Physical Evaluation:

The Poly herbal tea was tested to see how they looked, smelled, felt, and their overall state. This helps us understand their appearance and characteristics.

a) Color: The color of poly herbal tea was observed by visual examination. The color of powder is light green and when tea dipped in the water the color is slightly yellowish brown.

b) Odor: The odor of poly herbal tea was found to be Aromatic.

c) Taste: The taste of poly herbal tea was found to be Slightly Spicy, and Refreshing.

2) pH:

pH of prepared poly herbal tea was measured by using pH paper. The solution of poly herbal tea was prepared by using 50ml of Distilled water boiled at (100°C) and set aside. pH was determined in three times for solution and the average value was calculated.

Fig.6. pH of Poly herbal tea

3) Bulk Density:

Bulk density measures the mass of powder per unit volume, including void spaces. It is calculated by dividing the mass of the powder by its volume. This property helps assess how the powder behaves during handling, packaging, and storage.

Bulk Density = Mass of the powder/Bulk Volume

4) Tapped Density:

Tap density is the measurement of the maximum packing of powder after it has been tapped to eliminate air voids. It provides insight into the compaction characteristics of the powder.

Tapped Density = Mass of the powder/ Tapped Volume

5) Carr’s Index (Compressibility Index):

The Carr’s Index measures the flowability and compressibility of powders. It is calculated from the difference between the bulk density and the tap density, divided by the tap density. A higher Carr's Index indicates poorer flow properties.

Carr’s Index (%) = Tapped Density−Bulk Density/ Tapped Density* 100

6) Hausner Ratio:

The Hausner Ratio is the ratio of the tap density to the bulk density. It indicates the flow properties of the powder. A ratio of less than 1.2 suggests good flowability, while a ratio above 1.5 indicates poor flow properties.

Hausner’s Ratio = Tapped Density/ Bulk Density

7) Angle of Repose:

The angle of repose is the maximum angle at which a powder can be piled without collapsing. It is an indicator of the powder's flowability. Powders with a low angle of repose flow more freely.

Angle of Repose(θ) = Tan ^-1(h/r)

Fig.7.Angle of repose

9) Ash Value:

Ash value indicates the inorganic residue left after the burning of a sample. It provides an estimate of the mineral content and helps in evaluating the purity and quality of the powder.

Procedure: Clean and dry a crucible, then weigh it to determine the initial weight. Weigh about 2.5grams of the Poly herbal tea and place it in the crucible. Place the crucible with the sample in a muffle furnace and heat it at 550–600°C for 30min to 1hours to burn off the organic components, leaving behind the inorganic ash. After the combustion process, cool the crucible in a desiccator and then weigh it. The weight of the ash is 0.9gm.

Ash Value (%) = Weight of ash/ weight of sample*100

a) Determining Water-Insoluble Ash: Water-insoluble ash represents the portion of the total ash that cannot be dissolved in water. It is indicative of the more stable, non-soluble minerals present in the sample.

Procedure: After determining total ash is 0.9gm, transfer 0.45gm into a beaker. Add 50mL of distilled water. Boil for 5-10 minutes to dissolve soluble salts. Filter through Watmann filter paper to collect insoluble ash. Rinse the residue with hot distilled water. Dry in an oven or air dry, then return to the crucible. Cool in a desiccator and weigh the water-insoluble ash.

b) Determining Acid-Soluble Ash: The acid-soluble ash represents the portion of the total ash that can dissolve in a diluted acid (usually hydrochloric acid), leaving behind only those minerals that are resistant to acid digestion.

Procedure: Transfer 0.45gm ash into a beaker, add 50 mL of 1N HCl. Boil for 5-10 minutes to dissolve soluble inorganic components. Filter the mixture through Watmann filter paper to separate undissolved ash. Wash the residue with distilled water. Dry the residue in the oven or air dry. Cool in a desiccator and weigh the crucible to determine the acid-soluble ash.

10) Diffusion Time:

The time required for the tea bag dipped in water and produce or diffuse the essential material, color and taste.

Procedure: Boil water to 90-100°C. Prepare a transparent beaker or mug for steeping. Weigh the tea sachet for consistency. Place the tea sachet in the hot water and start the timer. Dipped if desired to help diffusion. Sample at intervals to check strength. Water should transition to a pale yellowish color. Record the steeping time (typically 1-2 minutes). Once desired strength is reached, remove the sachet to avoid over-brewing.

11) Stability Testing:

For herbal formulations to retain their identity, potency, quality, and efficacy over time, stability testing is an essential step in the development process. Stability testing guarantees that the formulation of polyherbal tea, which consists of a combination of different herbal powders, stays physically, chemically, and microbiologically stable under suggested storage settings²⁷.

12) Chemical Tests:

Table No.6.Chemical Test for Alkaloids

Alkaloids
Sr. No.	Test	Observation	Result
1	Hager’s Test	Yellow ppt	Present
2	Wagner’s Test	Reddish Brown ppt	Present

Table No.7. Chemical Test for Glycosides

Glycosides
Sr. No.	Test	Observation	Result
1	Molish Test	Voilet ring form between two layers	Present
2	Benedict Test	Green Color	Present
3	Killer Killani Test	Red ring form	Present

Table No.8. Chemical Test for Flavanoids

Flavanoids
Sr. No.	Test	Observation	Result
1	Lead Acetate	Yellow color	Present

Fig.8.Chemical Tests

13) Antioxidant Activity:

Preparation of phosphate buffer (pH 7.4):

Mix 2.38grams of disodium hydrogen phosphate, 0.19 grams of potassium dihydrogen phosphate, and 8grams of sodium chloride with enough water to generate a 1000 milliliter phosphate buffered saline solution with a pH of 7.4. To reach the appropriate level, adjust the pH as necessary. Because it is isotonic and non-toxic to the majority of cells, phosphate-buffered saline is frequently employed in biological research for a number of purposes, including substance dilution and cell rinsing²⁸.

Preparation of 40mM Hydrogen Peroxide:

To prepare a solution 0.445ml of hydrogen peroxide (H2O2) is dissolved in 100ml of phosphate buffer with a pH of 7.4.

Procedure: A solution of hydrogen peroxide (40 mM) was prepared in phosphate buffer (pH 7.4). Different concentrations (10, 20, 30, 40, and 50µg/mL) of the Guava leaf powder (or ascorbic acid as the control) were added to a hydrogen peroxide solution. The absorbance of hydrogen peroxide at 230nm was determined after 10 min against a blank solution containing phosphate buffer without hydrogen peroxide²³. The hydrogen peroxide scavengering activity of guava leaf derermine at 245nm. The hydrogen peroxide percentage scavenging activity was then calculated using the following equation:

Percentage of Scavenged (H₂O₂)

= (Abs Control-Abs sample)/(Abs control) *100

Antioxidant Activity:

a) Antioxidant activity of Guava leaf powder

Table No. 9. Radial scavengering activity of Guava Leaf Powder

Concentration	Absorbance value mean
Concentration	Control	Standard	Test
10 PPM	5.043	0.143	0.064
20 PPM	5.043	0.347	0.096
30 PPM	5.043	0.446	0.103
40 PPM	5.043	0.609	0.131
50 PPM	5.043	0.618	0.142

Table No.10.Percentage of scavengering activity

Concentration % inhibition	Absorbance value mean
Concentration % inhibition	Control	Standard	Test
10 ppm	0	97.16	98.73
20 ppm	0	93.11	98.09
30 ppm	0	91.15	97.95
40 ppm	0	87.92	97.40
50 ppm	0	87.74	97.18

Fig.9.Antioxidant activity of Guava Leaf Powder

The H2O2 assay was used to assess the antioxidant capacity of guava (Psidium guajava) leaf powder. At all concentrations, the test demonstrated consistently high radical scavenging activity, with values ranging from 98.73% at 10ppm to 97.18% at 50ppm. These readings showed superior antioxidant potential because they were marginally higher than the benchmark. According to the findings, guava leaf powder is a good source of natural antioxidants and might be used in products that fight oxidative stress²⁹.

b) Antioxidant activity of Poly herbal tea:

Table No.11. Radial scavengering activity of Poly herbal tea

Concentration	Absorbance value mean
Concentration	Control	Standard	Test
10 ppm	5.043	0.143	0.050
20 ppm	5.043	0.347	0.108
30 ppm	5.043	0.446	0.153
40 ppm	5.043	0.609	0.191
50 ppm	5.043	0.618	0.225

Table No.12. Percentage of scavengering activity

Concentration % inhibition	Absorbance value mean
Concentration % inhibition	Control	Standard	Test
10 PPM	0	97.16	99.00
20 PPM	0	93.11	97.85
30 PPM	0	91.15	96.96
40 PPM	0	87.92	96.21
50 PPM	0	87.74	95.53

Fig.10.Antioxidant activity of Poly herbal tea

The formulated Poly herbal tea exhibited strong antioxidant activity, with % inhibition ranging from 95.53% to 99.00%, comparable to ascorbic acid. This indicates the presence of potent phytoconstituents like flavonoids and polyphenols capable of effectively scavenging free radicals. Notably, high activity was observed even at low concentrations, highlighting the formulation's efficiency in reducing oxidative stress. These results support the poly herbal tea' potential as a natural antioxidant supplement for immune enhancement, with existing evidence on the antioxidant properties of Guava leaves.

6. CONCLUSION:

The present study on the formulation and characterization of polyherbal tea using guava leaf powder, lemongrass powder, tulsi powder, and ginger powder has demonstrated the potential of a natural, health-promoting beverage rich in bioactive compounds. Each ingredient was selected based on its traditional medicinal value and known phytochemical properties. The combination of these herbs resulted in a synergistic blend with enhanced organoleptic appeal and functional health benefits. Sensory evaluation indicated good acceptability in terms of taste, aroma, and color. Physicochemical analysis confirmed the presence of significant amounts of phenolic compounds and flavonoids, which contribute to antioxidant activity. The formulated polyherbal tea can serve as a natural remedy to improve immunity, aid digestion, and act as an antioxidant beverage. Moreover, the absence of artificial additives makes it a safe.

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Received on 15.08.2025 Revised on 13.10.2025

Accepted on 28.11.2025 Published on 30.01.2026

Available online from February 05, 2026

Res. J. Pharma. Dosage Forms and Tech.2026; 18(1):9-16.

DOI: 10.52711/0975-4377.2026.00002

This work is licensed under a Creative Commons Attribution-Non Commercial-Share Alike 4.0 International License. Creative Commons License.

Storage Condition	Room Temp.
Parameter	Initial	Week 1	Week 2	Week 3	Week4	Week 5
Color	Yellowish Brown	No Change	No Change	No Change	No Change	No Change
Odor	Aromatic	No Change	No Change	No Change	No Change	No Change
Taste	Spicy, Refreshing	No Change	No Change	No Change	No Change	No Change
pH	6 -7	No Change	No Change	No Change	No Change	No Change