INTRODUCTION:

The increasing interest in natural products as sources of bioactive compounds has driven extensive research into various fruits and plants. Guava (Psidium guajava L.), a tropical fruit, is renowned for its rich nutrient profile and medicinal properties, particularly in traditional medicine across various cultures. It is commonly used to treat ailments such as diarrhea, dysentery, and diabetes (Joseph and Priya, 2011). The pharmacological activities of guava are attributed to its high content of phytochemicals, including alkaloids, saponins, phenolic compounds, and terpenoids (Nantitanon et al., 2010).

Phytochemicals such as alkaloids are known for their therapeutic properties, including analgesic, anti-inflammatory, and anticancer activities (Cushnie et al., 2014). Saponins have been shown to possess antimicrobial, anti-inflammatory, and cholesterol-lowering properties (Vincken et al., 2007). Phenolic compounds, being potent antioxidants, play a significant role in protecting against oxidative stress-related diseases, including cardiovascular diseases and cancer (Shahidi and Ambigaipalan, 2015). Terpenoids, another major class of phytochemicals, are known for their diverse biological activities, including antimicrobial, anti-inflammatory, and anticancer effects (Gershenzon and Dudareva, 2007). The guava plant's leaves, in addition to its fruit, have been investigated for their potential health advantages. These benefits are attributed to the leaves' abundance of Phytochemical, which include guaijaverin, caffeic acid, catechin, quercetin, apigenin, kaempferol, myricetin, hyperin, gallic acid, epicatechin, chlorogenic acid, epigallocatechin gallate, and avicularin (Kumar et al., 2021). Guava extracts have been examined for their biological effects, including their hepatoprotective, ant diarrheal, anticancer, antioxidant, ant diabetic, and antibacterial properties (Porwal, Singh, and Gurjar, 2012).

The constituents in guava extracts have a variety of biological properties, including anti-inflammatory, hypoglycemic, anti-cancer, and antioxidant properties. Additionally, compared to unsulfated guava extract, sulfated guava extract exhibits greater bioactivities, including antioxidant, antimicrobial, and anticancer properties (Kumar et al., 2021). Despite the extensive use of guava in traditional medicine, there is limited systematic research on the phytochemical composition of guava extracts from different parts of the plant and using various solvents. This study aims to bridge this gap by conducting a comprehensive phytochemical analysis of guava fruits and leaves using ethanol, methanol, acetone, and aqueous extracts. The objective is to identify and quantify the major phytochemicals—alkaloids, saponins, phenolic compounds, and terpenoids—across these extracts, providing insights into the potential health benefits and applications of guava as a natural source of bioactive compounds.

MATERIALS AND METHODS:

Plant Material Collection:

Fresh guava (Psidium guajava L.) fruits and leaves were collected from a local orchard in Ch. Sambhajinagar during the peak harvest season. The plant material was authenticated by a botanist at Department of Botany, R. B. Attal College Georai- 431127, India and voucher specimens were deposited in the herbarium for future reference.

Preparation of Extracts:

The collected guava fruits and leaves were thoroughly washed with distilled water to remove any debris. The plant materials were then air-dried at room temperature (25±2°C) for two weeks until a constant weight was achieved. The dried samples were ground into a fine powder using a mechanical grinder. Approximately 50g of the powdered material was extracted with 500mL of solvents (ethanol, methanol, acetone, and distilled water) using a Soxhlet apparatus for 8hours (Harborne, 1998). The extracts were filtered through Whatman No. 1 filter paper and concentrated under reduced pressure using a rotary evaporator at 40°C. The concentrated extracts were stored in airtight containers at 4°C for further analysis.

Phytochemical Screening:

Qualitative phytochemical screening was conducted on the extracts to detect the presence of alkaloids, saponins, phenolic compounds, and terpenoids using standard procedures.

· Alkaloids: The presence of alkaloids was tested using Wagner's reagent, as described by Evans (2002). A few drops of the reagent were added to 2 mL of each extract, and the formation of a brown or reddish precipitate indicated the presence of alkaloids.

· Saponins: The froth test was employed to detect saponins. Briefly, 5mL of the extract was diluted with 20mL of distilled water and shaken vigorously for 15minutes. The formation of stable foam persisting for more than 15minutes confirmed the presence of saponins (Sofowora, 1993).

· Phenolic Compounds: The presence of phenolic compounds was assessed using the ferric chloride test. A few drops of 1% ferric chloride solution were added to 2mL of the extract. The appearance of a blue-black or dark green color indicated the presence of phenolics (Singleton et al., 1999).

· Terpenoids: The presence of terpenoids was tested using the Salkowski test. To 5mL of the extract, 2 mL of chloroform was added, followed by the careful addition of 3mL of concentrated sulfuric acid. The appearance of a reddish-brown interface indicated the presence of terpenoids (Siddiqui and Ali, 1997).

Quantitative Analysis:

Quantitative analysis of the detected phytochemicals was performed using spectrophotometric methods.

· Total Alkaloid Content: Alkaloid content was determined using the method described by Shamsa et al. (2008). The results were expressed as milligrams of atropine equivalents per gram of extract (mg AE/g).

· Total Saponin Content: The saponin content was estimated using the method by Obadoni and Ochuko (2001), and the results were expressed as milligrams of diosgenin equivalents per gram of extract (mg DE/g).

· Total Phenolic Content: The total phenolic content was determined using the Folin-Ciocalteu method, as described by Singleton et al. (1999). The results were expressed as milligrams of gallic acid equivalents per gram of extract (mg GAE/g).

· Total Terpenoid Content: Terpenoid content was quantified following the method by Ghorai et al. (2012), with the results expressed as milligrams of linalool equivalents per gram of extract (mg LE/g).

RESULTS:

The phytochemical analysis of guava (Psidium guajava L.) fruits and leaves revealed the presence of various bioactive compounds, including alkaloids, flavonoids, tannins, saponins, phenolic compounds, and terpenoids. The concentrations of these compounds varied depending on the solvent used for extraction. The results are summarized in Table 1 and 2.

Alkaloids were detected in both fruits and leaves across all extracts. The highest concentration of alkaloids was observed in the ethanol extracts, followed by methanol. Acetone and aqueous extracts contained lower concentrations. Flavonoids were found in significant amounts in the ethanol and methanol extracts. The acetone extracts exhibited a moderate concentration of flavonoids, while the aqueous extracts had the lowest levels.

Tannins were present in all extracts, with the highest concentration in the ethanol extracts. Methanol and acetone extracts showed moderate levels of tannins, while the aqueous extracts had the least. Saponins were detected in all extracts, with the aqueous extracts showing the highest concentration. The ethanol and methanol extracts had moderate levels of saponins, and the acetone extracts contained the lowest concentration. Phenolic compounds were abundant in the ethanol and methanol extracts. The acetone extracts exhibited moderate levels of phenolics, while the aqueous extracts had the lowest concentration. Terpenoids were detected in all extracts, with the highest concentration in the acetone extracts. Ethanol and methanol extracts had moderate levels of terpenoids, and aqueous extracts showed the lowest concentration.

Table 1: Phytochemical Composition of Guava (Psidium guajava L.) Fruits Extracts

Phyto-chemicals	Ethanol Extract	Methanol Extract	Acetone Extract	Aqueous Extract
Alkaloids	+++	+++	++	+
Flavonoids	+++	+++	++	+
Tannins	+++	++	++	+
Saponins	++	++	+	+++
Phenolic Compounds	+++	+++	++	+
Terpenoids	++	++	+++	+

Table 2: Phytochemical Composition of Guava (Psidium guajava L.) Leaves Extracts

Phytochemicals	Ethanol Extract	Methanol Extract	Acetone Extract	Aqueous Extract
Alkaloids	++	++	++	+
Flavonoids	+++	+++	++	++
Tannins	+++	++	++	+
Saponins	++	++	+	+
Phenolic Compounds	+++	++	++	+
Terpenoids	++	+	+	+
Where, Low: +, Moderate: ++ and High: +++

DISCUSSION:

The present study aimed to evaluate the phytochemical composition of guava (Psidium guajava L.) fruits and leaves extracted using different solvents, including ethanol, methanol, acetone, and water. The results revealed significant variability in the concentrations of alkaloids, flavonoids, tannins, saponins, phenolic compounds, and terpenoids, depending on the extraction solvent used. These findings underscore the importance of solvent selection in maximizing the yield of specific bioactive compounds, which is crucial for the potential application of guava in nutraceuticals, pharmaceuticals, and functional foods.

Alkaloids were detected in all extracts, with the highest concentrations found in the ethanol extracts of both guava fruits and leaves. This is consistent with previous studies that have demonstrated the efficacy of ethanol in extracting alkaloids due to its polar nature, which facilitates the solubilization of these compounds (Cushnie et al., 2014). The presence of alkaloids in significant amounts highlights the therapeutic potential of guava, particularly in treating conditions such as pain, inflammation, and infections (Gurib-Fakim, 2006).

Flavonoids, known for their strong antioxidant properties, were most abundant in the ethanol and methanol extracts. This result aligns with previous research, indicating that these solvents are effective in extracting flavonoids due to their ability to disrupt plant cell walls and release these compounds (Kaurinovic and Vastag, 2019). The antioxidant potential of flavonoids suggests that guava extracts, particularly those obtained with ethanol and methanol, could be valuable in preventing oxidative stress-related diseases, such as cardiovascular diseases and certain types of cancer (Shahidi and Ambigaipalan, 2015).

The highest concentrations of tannins were observed in the ethanol extracts. Tannins are well-known for their astringent properties and their ability to precipitate proteins, which contributes to their use in traditional medicine for treating diarrhea and wound healing (Haslam, 1996). The presence of high levels of tannins in the ethanol extracts of guava supports its traditional use in treating gastrointestinal disorders and suggests potential applications in modern medicine.

Saponins were detected in all extracts, with the highest concentration found in the aqueous extracts. This finding is consistent with the hydrophilic nature of saponins, which makes water an effective solvent for their extraction (Vincken et al., 2007). Saponins have been shown to possess various biological activities, including antimicrobial, anti-inflammatory, and cholesterol-lowering effects (Man et al., 2010). The high concentration of saponins in guava extracts, particularly in aqueous extracts, suggests that guava could be explored as a natural source of these bioactive compounds for therapeutic applications.

Phenolic compounds were abundant in both ethanol and methanol extracts, which is in line with previous studies that have reported the efficiency of these solvents in extracting phenolics (Dai and Mumper, 2010). Phenolic compounds are known for their antioxidant, anti-inflammatory, and anticancer properties (Shahidi and Ambigaipalan, 2015). The high phenolic content in guava extracts indicates their potential for use in the development of antioxidant-rich products that could help in managing oxidative stress-related conditions.

Terpenoids were found in all extracts, with the highest concentration in the acetone extracts. Acetone's effectiveness in extracting terpenoids is attributed to its ability to dissolve both polar and non-polar compounds, making it suitable for isolating these bioactive molecules (Gershenzon and Dudareva, 2007). Terpenoids have been recognized for their antimicrobial, anti-inflammatory, and anticancer activities (Nisar et al., 2018). The significant presence of terpenoids in guava extracts, particularly in acetone extracts, suggests that guava could be a valuable source of these compounds for therapeutic use.

The variability in phytochemical content across different solvents highlights the need for targeted extraction strategies based on the desired bioactive compounds. Ethanol and methanol are particularly effective for extracting alkaloids, flavonoids, phenolic compounds, and tannins, while water is more suitable for saponins, and acetone excels in terpenoid extraction. These findings can inform the development of guava-based products tailored for specific health benefits.

CONCLUSION:

Future research should focus on the bioactivity and pharmacokinetics of these phytochemicals to better understand their therapeutic potential and safety. Additionally, in vivo studies are needed to validate the efficacy of guava extracts in preventing or treating various diseases, thereby bridging the gap between traditional medicine and modern therapeutics.

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Received on 28.02.2025 Revised on 17.03.2025

Accepted on 02.04.2025 Published on 10.05.2025

Available online from May 14, 2025

Res. J. Pharmacognosy and Phytochem. 2025; 17(2):107-110.

DOI: 10.52711/0975-4385.2025.00018

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