INTRODUCTION:

One of the most prevalent concerns among men facing sexual health issues is erectile dysfunction (ED)^1–3. Erectile dysfunction is a substantial medical condition affecting a significant portion of the male population, ranging from 10% to 52%⁴. Notably, men tend to experience a higher prevalence of sexual disorders compared to women.

These disorders encompass a spectrum including diminished libido, ED, premature ejaculation, retrograde ejaculation, and discomfort during sexual intercourse^5,6. The global incidence of ED is anticipated to escalate, with projections indicating that by 2025, it will impact an estimated 322 million men worldwide, marking a substantial 111% surge from 1995⁷.

Aphrodisiacs, as defined, are organic compounds or medications known for their capacity to enhance sexual arousal^8,9. Plants belonging to the Piperaceae family, characterized by the presence of essential oils⁸, as well as those from the Solanaceae family containing flavonoid antioxidants and the Cucurbitaceae family with amino acids, exhibit substantial potential as aphrodisiac agents^10–12 Additionally, certain foods endowed with antioxidant properties can also serve as aphrodisiacs. These include chili peppers, basil leaves, purple eggplant, strawberries, watermelon, raw oysters, chocolate, coffee, and honey^10,11,13,14.

These natural components comprise active compounds representing various classes, including alkaloids, essential oils, flavonoids, and terpenoids, which possess the capability to stimulate androgen hormones. Androgen hormones, when activated, induce masculinizing effects, stimulate spermatogenesis in the testes, enhance aggression, contribute to bone and muscle growth, and influence sexual behavior¹⁵. Alkaloids were found in some of the plants developed in this study, including the piperine from red chilli, citrulline in the Watermelon albedo. Other alkaloids such as tropane, pyrrolidine, quinazolizidine are found in the Eggplant fruit¹⁶. Basil leaf contains arginine that can enhance sperm movement and prevent infertility contains arginine, which increases the movement of sperm and prevents infertility¹⁷. The flavonoids contained in red gingger have a role in increasing libido through increased levels of dehydration¹⁸. The primary objective of this research is to investigate diverse plant sources that can be harnessed for the preservation of reproductive health. This endeavor aims to harness natural resources and promote local specialties within Indonesia.

MATERIALS AND METHODS:

Materials:

The materials utilized in this research comprised leaves from Piper retrofractum Vahl, Solanum melongena egg plant, Ocimum basilicum leaves, Zingiber officinale Var Rubrum rhizoma, Watermelon albedo. These plant specimens were sourced from Central Java, Indonesia. Additionally, the chemicals employed included sildenafil citrate, Na-CMC (sodium carboxymethyl cellulose) at a concentration of 0.05%, 70% Ethanol, and distilled water.

Extract Preparation:

Simplisia were soaked in Ethanol 70% for a duration of one day, maintaining a ratio of 1 part powder to 10 parts Ethanol 70%. The resulting filtrate was subjected to evaporation using a vacuum evaporator for a period of 6 hours, followed by further drying with a dry exhauster for 24 hours, ultimately yielding a concentrated, thick extract.

Phytochemical Analysis of The Extracts:

The extracts were subjected to screening to detect the presence of various compounds including alkaloids, flavonoids, tannins, steroids, terpenoids, and saponins¹⁵, as summarized in Table 1.

Table 1: Methods and observation of the active components of various extracts

Components	Method	Observation
Alkaloids	Extract + 1% HCl 2N+ NaCl + Mayer’s reagent	Formed yellowish white precipitate on Meyer’s reagent
Flavonoids	Extract (2mL; 0.5g/5mL) + Mg + 1mL HCl + 5mL amyl alcohol	Formed red, yellow or orange color
Tannins	Extract (2mL; 0.5g/5 mL) + 5mL hot distilled water 3mL NaCl 10% and gelatin 1%.	Precipitate formed
Saponins (Froth’s Test)	Extract (1mL; 0.5g/5mL) + water (5mL)	Forms a stable foam with a height of 1-10 cm for 10 minutes
Terpenoids (Salkowki’s Test)	Extract (2 mL; 0.5 g/5 mL) + chloroform (1 mL) + conc. H₂SO₄ (2 mL)	Red brownish precipitate

Preparation of Test Animals:

Male and female white rats were utilized as experimental subjects and were individually housed in cages measuring approximately 50 x 30 cm. The room temperature was maintained at 2±1℃, and lighting conditions followed a 12-hour light-dark cycle. During a 7-day acclimation period, the rats received standard feed once daily and had unrestricted access to drinking water¹⁹. Ethical approval for the research protocol was obtained from the Health Research Ethics Commission of the Faculty of Medicine, Universitas Muhammadiyah Surakarta, under Ethics Decree Number 4715/A.1/KEPK-FKUMS/II/2023.

Treatment of test animals for aphrodisiac activity:

A total of 60 male rats were allocated into 12 separate groups, and each group underwent a 7-day treatment regimen as follows:

Group 1 received 10ml/kgbw of distilled water (Negative Control/NC).

Group 2 received sildenafil citrate (SD) at a dosage of 10mg/kgbw (positive control).

Groups 3-7 were administered test preparations, with each receiving an extract dose of 10mg/kgbw

Groups 8-12 were administered test preparations, with each receiving an extract dose of 30mg/kgbw

On the seventh day of the experiment, a group comprising one male and three female white rats was introduced into an observation enclosure. The observation sessions were conducted during the evening hours, precisely from 19:00 to 22:00, each session lasting for a duration of one hour. The camera was used to observe the sexual behavior of the test animals on the parameters of frequency of introduction, climbing, and coitus. Introduction is the initial approach process in which the male rat approaches the female rat and begins to kiss the female rat's body until licking the female rat's genitals and rectum. Climbing occurs when male rats begin to ride or climb the body of female rats prior to ejaculation. The activity of coitus occurs when the male rat climbs the female rat's body until ejaculation occurs. Male rats lick their own genitals after ejaculation. This study's observations are based on three parameters: introduction observations, climbing observations, and coitus observations^15,19.

The research study documented the variables of mating initiation, locomotion, and copulation by quantifying the frequency at which a male rat engaged in these behaviors with a female rat. The present study focuses on the sexual behavior exhibited by male rats, namely the act of approaching the female rat and engaging in oral contact with her mouth and rectal area. In the context of human psychology, the concept of introduction pertains to the manifestation of libido. The behavior is recorded when the male rat engages in approach behavior towards the female rat and engages in physical contact with her body parts. In the context of rat behavior, climbing is a term used to describe a sexual action in which male rats engage in mounting the female rat from a posterior position. In the context of human physiology, the term "climbing" pertains to the temporal aspect of the ejaculation process. The duration of climbing is quantified from the initiation of the male rat's ascent onto the female rat's body. The duration of climbing is quantified as the time elapsed from the initiation of the male rat's ascension onto the female rat's body. The duration of climbing is quantified as the time elapsed from the initiation of the male rat's ascension onto the female rat's body. Coitus refers to the sexual behavior exhibited by male and female rats. During copulation, male rats participate in sexual intercourse and afterwards engage in self-grooming behaviors, whereas female rats exhibit body stretching and tail lifting. In the context of human biology, the term "coitus" is used to describe sexual intercourse. It is possible to assess the duration of coitus by evaluating an individual's stamina throughout the mating process²⁰.

Histopathology test.

On the 7th day the test animal was sacrificed with euthanasia using a 10:1 mixture of ketamine 100 mg/ mL and xylazine 100mg/mL. The testicular organs are examined using a standard formalin buffer solution (NBF) and stained with hematoxylin-eosin (HE). Each preparation is prepared 3 slices and observed visibility of cells experiencing spermatogonia, spermatocytes, spermatids under a microscope with 1000x magnification. Quantification of spermatogonia, spermtocytes, and spermatids is conducted across the whole seminiferous tubules present in a testis sample, which can be visualized and quantified by examining five tubules. The outcome of the counting is obtained by dividing the total number of cells by the quantity of observed tubuli, and the resulting value is presented as an average.

Data Analysis:

The collected data were visually represented through graphical illustrations and tabular formats. Data analysis was conducted using the Statistical Package for the Social Sciences (SPSS), specifically version 26 for Windows. The statistical tests employed in the analysis included the Kruskal-Wallis test and the Mann-Whitney test to assess significant differences and associations within the dataset.

RESULTS:

Extraction Result:

The extraction process utilizing suitable solvents yielded extracts with satisfactory yields and the detection of compounds likely to contribute to the intended effects²¹. The extraction procedure was conducted using appropriate organic solvents, resulting in the production of concentrated and desiccated extracts. The identification of chemical compounds present within the extracts was performed via tube tests using suitable reagents and corresponding detection methods, as detailed in Table 2. It was observed that all the extracts contained notable quantities of flavonoids, alkaloids, and saponins.

Alkaloids, flavonoids, and saponins were present in all of the extracts examined^9,22,23. The alkaloids piperine and piperidin⁸ are found in PRE. Eggplant contains several alkaloids, including tropane, pyrrolidine, quinazolizidine, steroid alkaloids, and glycoalkaloids²². Watermelon albedo extract (WAE) featured a significant alkaloid, citrulllin, known for its potential in improving erectile function^24,25. Red ginger (ZRE) was characterized by a rich composition of phenolic compounds, including gingerol, paradols, and shogaol²³. The essential oil of O. basilicum contained eucalyptol (1.79%), linalool (12.63%), terpineol (0.95%), and eugenol (19.22%)²⁶.

Results of Aphrodisiac activity in experimental animals:

The frequency of introduction (FI) of aphrodisiacs, or drugs capable of boosting libido or sexual excitement, can be used to measure their impact on male rat libido. FI measures the number of times male rats approach and engage with female rats, indicating higher libido²⁰. Only the PRE extract did not cause a significant increase in libido when provided at a dose of 10mg/kgbw. However, when each extract was delivered at a dose of 30mg/kg bw, libido increased significantly (p<0.05) compared to the NC group, however this augmentation did not replicate the effect seen with SD 10(Table 3).

Table 3: Frequency of occurrence of Introduction, Climbing, and Coitus parameters

Treatment groups	Frequency of occurence (times)
Treatment groups	Introduction (FI)	Climbing (FCl)	Coitus(FCo)
NC	22.20 ±5.64	16.80±3.97	13.40 ±3.72
SD 10	70.40 ± 9.16	65.60 ± 10.05	53.20 ±10.65
PRE 10	31.20 ± 4.12	23.00 ± 3.46	19.40 ± 3.07^.*
PRE 30	50.20 ± 7.47^.*	41.00 ± 7.16	37.00 ± 5.02
EE 10	39.80 ± 4.86^.*	34.00 ± 5.14^.*	22.80 ±3.42^.*
EE 30	65.60 ± 8.35^*.#	56.40 ± 11.58^*.#	44.20 ±14.29^*.#
OBLE 10	41.60 ± 10.92^.*	34.00 ± 10.79^.*	28.00 ± 10.49^.*
OBLE 30	56.00 ± 8.06^.*	49.20 ± 8.67^.*	45.00 ± 9.30^*.#
WAE 10	28 ± 2.83^**	23.2 ± 2.79^.*	19 ± 2.45^.*
WAE 30	44.60± 7.47^.*	37.20 ± 8.03^.*	26.2 ± 4.58^.*
ZRE 10	37.80 ± 9.33^.*	33.20± 9.06^.*	31.20 ± 8.52^.*
ZRE 30	64.60 ± 13.66^*.#	60.20 ± 14.33^*.#	51.60 ± 14.15^*.#

Notes: *: different with NC (p<0.05), **: different with SD 10 (p<0.05), #: not significantly different with SD 10 (p>0.05)

The evaluation of ejaculation behavior can be deduced from the climbing frequency parameter (FCl), which signifies the stage in which male rats mount female rats during mating. FCl offers insights into the duration of male ejaculation or pre-copulatory behaviors²⁰. Notably, administration of PRE did not induce a significant change in this behavior (p>0.05), whereas the other extracts, both at doses of 10 and 30mg/kg bw, exhibited an increase. Remarkably, administration of EE and ZRE at a dose of 30mg/kg bw demonstrated similar effects to SD 10(p>0.05) (Table 2).

Coitus encompasses the sexual activities of male rats, involving several stages such as male rats licking their own genitals and female rats extending their bodies and raising their tails. Among all the treatments, only the 30 mg/kg bw dose of PRE did not result in a significant increase in coitus frequency (p>0.05 compared to NC). Conversely, administration of EE, OBLE, and ZRE exhibited coitus frequency (FCo) equivalent to that of SD 10(p<0.05) (Table2).

The testicles, crucial reproductive organs responsible for sperm production and sex hormone secretion, comprise seminiferous tubules, connective tissue, and blood vessels. The seminiferous tubules represent the primary component of the testicles, and any damage or atrophy to the cells within them can lead to a reduction in testicular weight. Aberrations in the testicles can potentially affect spermatogenesis, impacting sperm quality. Testicular weight can be determined by weighing each individual testis, serving as an effective indicator of cell growth activity and endocrine secretion levels at the fifteen-minute mark²⁰.

Table 4: The weights (grams) of animal tests at the end of the study (H-7), right and left testicular organ weights (grams) and relative organ weights (%).

Treatment groups	Rat weight on day 7	Weight of right + left testicle	Relative weight of organs (%)
NC	239.6 ± 10.53	2.44 ± 0.21	1.02 ± 0.11
SD 10	207.8 ± 13.12	2.55 ± 0.36	1.24 ± 0.23
PRE 10	228.2 ± 14.01^**	2.33 ± 0.13^*	1.02 ± 0.04^**
PRE 30	217.8 ± 27.41^.*	2.54 ± 0.121^*.#	1.18 ± 0.12^*.#
EE 10	211.0 ± 5.09^*	2.25 ± 0.04^**	1.06 ± 0.02^**
EE 30	208.6 ± 4.22^*.#	2.28 ± 0.05^**	1.09 ± 0.03^**
OBLE 10	230.8 ± 10.70^**	2.43± 0.08^**	1.05 ± 0.04^**
OBLE 30	196.2 ± 13.36^*	2.27 ± 0.11^**	0.97 ± 0.48^**
WAE 10	226.2 ± 25.78^.*	2.29 ± 0.11^**	1.02 ± 0.11^**
WAE 30	207.4 ± 9.68^*.#	2.12 ± 0.19^**	1.01 ± 0.11^**
ZRE 10	222.6 ± 16.28^.*	2.36 ± 0.21^**	1.062 ± 0.10^**
ZRE 30	222.4 ± 21.66^.*	2.38 ± 0.11^**	1.082 ± 0.10^**

Notes: *: different with NC (p<0.05), **: different with SD 10 (p<0.05), #: not significantly different with SD 10 (p>0.05)

The connection between the relative testicular weight and aphrodisiac effects is characterized by the correlation between higher testosterone hormone production in the testes, increased testicular weight, and enhanced sexual behavior, as reflected in elevated FI, FCl, and FCo²⁴. The outcomes revealed that the % relative organ weight following the administration of SD 10 was comparable to that observed with PRE at 30 mg/kgbw (p>0.05) (Table 4).

Histopathologic features of testicular organs after treatment:

Histopathological assessments of testicular organs following treatment revealed the presence of spermatogonia, a diverse group of proliferating cells within the testis, representing early stages of sperm maturation (see Figure 1). Spermatogonia undergo mitosis, giving rise to preleptotene spermatocytes or resting primary spermatocytes, which subsequently enter an extended meiosis phase of spermatogenesis, lasting approximately 24 days in humans. During meiosis, primary spermatocytes divide into secondary spermatocytes, leading to a reduction in chromosome number. Spermiogenesis signifies the stage in which round spermatids transform in shape and composition to assume a tadpole-like structure, featuring a head, neck, tail, and motility. In the maturation phase, spermatids acquire a morphology nearly identical to mature spermatozoa, detaching from the seminiferous epithelium to become free spermatozoa²⁷.he histopathological analysis indicates that all extracts administered at doses of 10 or 30 mg/kg bw exhibit significant differences from the negative control (p<0.05). However, the cellular morphology resulting from these extracts does not closely resemble that induced by sildenafil citrate (p<0.05), as outlined in Table 5.

Table 5: Histopathological features of testicular organs in terms of the number of cells in the Spermatogonia, Spermatocid, Spermatid phase

Treatment groups	Spermatogonia	Spermatocid	Spermatid
NC	41.0 ± 1.41	56.6 ± 5.12	31.8 ± 4.92
SD 10	62.0 ± 2.83	144.6 ± 4.41	72.6 ± 6.37
ECB 10	50.2 ± 6.08^.*	81.6 ± 6.83^.*	40.6 ± 5.95^.*
ECB 30	54.0 ± 2.83^.*	107.2 ± 9.11^.*	53.8 ± 4.49 ^.*
EE 10	45.2 ± 3.54^**	70.6 ± 6.77^.*	36.8 ± 4.71^**
EE 30	48.4 ± 1.85^.*	109.4 ± 5.89^.*	45.2± 3.54^.*
OBLE 10	45.6 ±2.06^**	66.2 ±12.01^.*	44.8 ± 2.86^.*
OBLE 30	51.0 ± 3.03^.*	100.6 ± 6.02^.*	48.4 ± 6.59^.*
WAE 10	52.5 ± 2.69^.*	66.0 ± 4.12^.*	41.0 ±1.87^.*
WAE 30	54.2 ± 1.72^.*	97.2 ± 7.49^.*	55.6 ± 5.12^.*
ZRE 10	48.4 ± 2.58^.*	64 ± 2.97^**	41.4 ± 2.06^.*
EHM 30	48.4 ±1.85^.*	93.2 ±8.61^.*	47.8 ±4.96^.*

Notes: *: different with NC (p<0.05), **: different with SD 10 (p<0.05), #: not significantly different with SD 10 (p>0.05)

The etiology of erectile dysfunction (ED) in are complex, involving multiple factors including vascular and neurological impairments. ED have compromised relaxation of the smooth muscle in the corpus cavernosum when exposed to nitric oxide produced from neuronal and endothelial sources. This impairment could potentially be attributed to the increase of glycosylation products²⁸. Sildenafil citrate, serving as a positive control, constitutes an active component employed in the oral treatment of erectile disorders. It functions through the inhibition of phosphodiesterase-5 (PDE-5), an enzyme specifically associated with cyclic Guanosine Monophosphate (cGMP)²⁹. During the course of sexual stimulation, the release of neurotransmitters, facilitated by nitric oxide (NO) within the corpus cavernosum, leads to an elevation in the levels of cGMP. This heightened cGMP concentration instigates the relaxation of muscles within the corpus cavernosum, thereby facilitating an increased blood flow into this region and subsequently engendering penile erection. The presence of phosphodiesterase-5 can impede the actions of cGMP. Consequently, sildenafil citrate, through its inhibition of phosphodiesterase-5, prolongs the activity of cGMP, enhancing the likelihood of achieving an erection when subjected to sexual stimulation³⁰.

Herbal medications can be utilized not only as a cure but also as growth boosters and preventive acts³¹. Alkaloids, with their inherent capacity to induce vasodilation, play a pivotal role in promoting erection by augmenting blood vessel dilatation. Piperine, a particular type of alkaloid, exerts influence over the reproductive hormone system by elevating the levels of gonadotropin hormone. Moreover, piperine exhibits stimulant and vasodilatory properties, thereby amplifying blood circulation to the reproductive organs^32,33. Another noteworthy alkaloid, citrulline, possesses polar characteristics and solubility in aqueous compounds. Citrulline exerts an impact on the production of potent vasodilators such as nitric oxide (NO)^34,35.

Flavonoid compounds play a crucial role in elevating dehydroepiandrosterone (DHEA) levels, which in turn have a significant impact on increasing testosterone levels and stimulating sexual activity in men. DHEA contributes to testosterone biosynthesis, with the enzymatic conversion of pregnenolone to the intermediary compound 17-hydroxypregnenolone mediated by CYPC17. Subsequently, cytochrome CYPC17 further transforms the intermediary product into dehydroepiandrosterone. Under the influence of the 3β-HSD enzyme, dehydroepiandrosterone is subsequently converted to androstenedione, ultimately leading to the formation of testosterone²⁶.

In addition to their central actions, these flavonoids and alkaloids also exert peripheral effects that contribute to the relaxation of the corpus cavernosum, facilitating penile erection²⁷. The central mechanisms involving alkaloids stimulate the release of nitric oxide from nerve endings and endothelial cells³⁰. The release of nitric oxide is triggered by erotic stimulation of the sexual center in the brain, with signals transmitted through parasympathetic nerve activation and signal transduction in the sacral erectile center within the corpus cavernosum. Key transmitters involved in this process include vasoactive intestinal polypeptide and nitric oxide, with nitric oxide serving as the principal mediator of corpus cavernosum relaxation³⁴. Nitric oxide plays a pivotal role in increasing cyclic Guanosine Monophosphate (cGMP) levels, which exert vasodilatory effects³². cGMP serves as a critical molecule that regulates various aspects of the erection process. It governs smooth muscle function and blood flow within the penile region, both crucial for initiating and sustaining an erection. Additionally, cGMP inhibits the activity of phosphodiesterase type 5 (PDE5), the enzyme responsible for cGMP breakdown. By inhibiting PDE5, cGMP levels remain elevated in erectile tissue, prolonging smooth muscle relaxation and ensuring sufficient blood flow to maintain a robust erection. This inhibition of PDE5 aligns with the working principles of conventional drugs such as sildenafil and tadalafil^36,37.

In addition to flavonoids and alkaloids, saponins also exhibit aphrodisiac properties. Saponins operate by enhancing libido and play a pivotal role in the biosynthesis of dihydrotestosterone, a key factor in elevating testosterone levels³⁸. Furthermore, saponins stimulate androgen production within the central nervous system and gonadal tissues, thereby directly or indirectly augmenting libido. Saponins also participate in the biosynthesis of dehydroepiandrosterone (DHEA), further contributing to the stimulation of libido and testosterone production²⁷. The surge in hormone levels has been correlated with heightened sexual desire, increased penile tumescence, rigidity, and the utilization of accessory muscles that contribute to sexual activity³⁹. Terpenoids, on the other hand, operate through a mechanism involving the binding of saponins to form a terpenoid-saponin bond. This interaction impacts sexual activity by replacing cholesterol in the synthesis of testosterone. Steroids, in a similar vein, exert their influence on sexual activity by substituting cholesterol in the process of testosterone synthesis. This occurs through the conversion of pregnenolone, a cholesterol precursor, into progesterone, a precursor that triggers the formation of androgens such as testosterone⁴⁰. Furthermore, aside from its central effects, testosterone has been observed to exert peripheral actions by enhancing nitrergic neurotransmission, amplifying the activity of nitric oxide synthase, and facilitating the release of nitric oxide within the cavernosa. Collectively, these mechanisms contribute to the intricate process of penile erection.

CONCLUSION:

The administration of 30 mg of ethanolic extract (EE) resulted in the highest aphrodisiac activity in the test animals, as evidenced by increased frequencies of introduction, climbing, and coitus. Notably, the treatment with PRE at 30 mg also led to a significant increase in the relative testes weight, resembling the effect observed with sildenafil at a dosage of 10 mg/kgbw.

CONFLICT OF INTEREST:

The authors declare no conflicts of interest with respect to this research.

ACKNOWLEDGMENTS:

The authors extend their appreciation to the Ministry of Education, Culture, Research, and Technology of the Republic of Indonesia for providing support through grant contract agreement number 006/LL6/PB/AL.04/2023, 170.18/C.1-III/LRI/VI/2023. Furthermore, special thanks go to Diaz Ivanda Ayu Dewayani and Vidya Nareta Deva Aryani, UMS Pharmacy students, for their assistance with this research. Muhammad Da’i is responsible for this project.

REFERENCES:

1. Situmorang EW, Kurniawati E, Soleha TU, Larasati T. Obesity and erectile dysfunction: Literature Review. PREPOTIF J Kesehat Masy. 2022; 6: 1258-1265. doi:10.31004/prepotif.v6i2.4336

2. Junairiah, Arofah J, Manuhara YSW, Nurhariyati T, Ni’matuzahroh. Effect of 2,4-d and bap on callus induction of piper retrofractum vahl. Res J Pharm Technol. 2021; 14(3): 1390-1394. doi:10.5958/0974-360X.2021.00248.1

3. Nagamani, K. Sesha Kumar, N. Sucharitha PVS. Male Sexual Dysfunction – An Over Looked Health Issue. A V Pub J Nurs Med Res. 2022; 1(1): 27-31. doi:10.52711/jnmr.2022.06

4. Singh R, Singh S, Jeyabalan G, Ali A, Semwal A. Sexual Dysfunction An Overview and Medicinal Plant used for treatment of Sexual dysfunction. Earthjournals Publ. 2012; 1(2): 9-24.

5. Luthfi MJ, Noor MM, Latip J. Medicinal Plants: A Prospect in Developing Male Fertility Enhancing Agent. Biol Med Nat Prod Chem. 2015; 4(2): 41. doi:10.14421/biomedich.2015.42.41-47

6. Kotta S, Ansari S, Ali J. Exploring scientifically proven herbal aphrodisiacs. Pharmacogn Rev. 2013; 7(13): 1-10. doi:10.4103/0973-7847.112832

7. Goldstein I, Goren A, Li VW, Tang WY, Hassan TA. Epidemiology Update of Erectile Dysfunction in Eight Countries with High Burden. Sex Med Rev. 2020; 8(1): 48-58. doi:10.1016/j.sxmr.2019.06.008

8. Wahyuni AS, Wahyuningtyas N, Arifiyanti. Aphrodisiac Activity Of Essential Oil From Syzygium aromaticum (L.) Merr. & Perry. Pharmacon. 2010; 11(2): 43-46.

9. Salleh MNH, Ahmad F. Phytopharmacological investigations of piper retrofractum vahl. – A review. Agric Conspec Sci. 2020; 85(3): 193-202.

10. Fajriyatun BN, Sutrisna E, Nirlawati DD. Investigating the Effects of 70% Etanol Extract from Oak Seeds (Solanum Melongena L.) on HDL Cholesterol Levels Male White Rat (Rattus Norvegicus) HDL Blood Serum Galur Wistar. Biomedika. 2014; 6(1): 13-16. doi:10.23917/biomedika.v6i1.282

11. Parisa N, Kamaluddin MT, Salni S. Aphrodisiac Effects of Areca Fruit in Erectile Dysfunction Rat Model. Biosci Med J Biomed Transl Res. 2019; 3(3): 16-23. doi:10.32539/bsm.v3i3.90

12. Dhanalakshmi S, Abinaya, Devi K, Lakshmi. In vitro anti-oxidant study of herbal extract mixture by nitric oxide and DPPH method. Res J Pharm Technol. 2017; 10(1): 277-280. doi:10.5958/0974-360X.2017.00057.9

13. Nurwaini S, Wikantyasning E. Formulations of Lozenges of Leaf Extract Ocimum sanctum L : The effect of sodium carboxymethyl cellulose as a binding substance on physical properties. J Penelit Sains Teknol. 2011; 12(1): 45-57. http://publikasiilmiah.ums.ac.id/handle/123456789/3434

14. Sharma R, Kumari N, Ashawat M., Verma CP. Standardization and Phytochemical Screening analysis for Herbal Extracts: Zingiber officinalis, Rosc., Curcuma longa Linn., Cinnamonum zeylanicum Nees., Piper longum, Linn., Boerhaavia diffussa Linn. Asian J Pharm Technol. 2020; 10(3): 127-133. doi:10.5958/2231-5713.2020.00022.7

15. Sharma M, Arya D, Bhagour K, Gupta RS. Natural aphrodisiac and fertility enhancement measures in males: A review. Curr Med Res Pract. 2017; 7(2): 51-58. doi:10.1016/j.cmrp.2017.02.007

16. Pantuzza Silva GF, Pereira E, Melgar B, et al. Eggplant fruit (Solanum melongena l.) and bio‐residues as a source of nutrients, bioactive compounds, and food colorants, using innovative food technologies. Appl Sci. 2021; 11(151): 1-24. doi:10.3390/app11010151

17. Zaini BSI, Pujiana IM, Djamaludin EO, Salim HM. The Effectiveness of Extract Basil Leaves on Motility and Morphology of Spermatozoa Mice Exposed with Cigarette Smoke. Biomol Heal Sci J. 2020; 3(2): 75. doi:10.20473/bhsj.v3i2.22169

18. Anandita DW, Nurlaila, Pramono S. Aphrodisiac Effects Of Red Ginger (Zingiber officinale Rosc. red clone) Essential Oil And Essential Oil Free Etanolic Extract In Male Rats. Maj Obat Tradisional,. 2012; 17(1): 8-14.

19. Wolfensohn S, Lloyd M. Handbook of Laboratory Animal Management and Welfare. Blackwell Scientific Publications Oxford and Edinburg; 2003. doi:10.1016/s0165-6147(00)89011-2

20. Tachikawa KS, Yoshihara Y, Kuroda KO. Sexual behavior and aggression in male mice: Involvement of the vomeronasal system. J Neurosci. 1984; 4(9): 2222-2229. doi:10.1523/jneurosci.04-09-02222.1984

21. Pal RS, Saraswat N, Pal Y, Wal P, Wal A, Rai AK. Alcoholic extract of poly herbal powder mixture for anti-obesity effect on wistar rats. Res J Pharm Technol. 2019; 12(4): 1857-1864. doi:10.5958/0974-360X.2019.00310.X

22. Islam MT. Phytochemical information and pharmacological activities of Okra (Abelmoschus esculentus): A literature-based review. Phyther Res. 2019; 33(1): 72-80. doi:10.1002/ptr.6212

23. Supu RD, Diantini A, Levita J, et al. Red Ginger ( Zingiber officinale var. rubrum): Its Chemical Constituents, Pharmacological Activities And Safety. Fitofarma. 2018; 8(1): 25-31.

24. Assefa AD, Hur OS, Ro NY, et al. Fruit morphology, citrulline, and arginine levels in diverse watermelon (Citrullus lanatus) germplasm collections article. Plants. 2020; 9(9): 1-16. doi:10.3390/plants9091054

25. Srivatsav A, Balasubramanian A, Pathak UI, et al. Efficacy and Safety of Common Ingredients in Aphrodisiacs Used for Erectile Dysfunction: A Review. Sex Med Rev. 2020; 8(3): 431-442. doi:10.1016/j.sxmr.2020.01.001

26. Khair-ul-Bariyah S, Ahmed D, Ikram M. Ocimum Basilicum: A Review on Phytochemical and Pharmacological Studies. Pakistan J Chem. 2012; 2(2): 78-85. doi:10.15228/2012.v02.i02.p05

27. Igwebuike UM, Ochiogu IS, Ihedinihu BC, Ikokide JE, Idika IK. The effects of oral administration of monosodium glutamate (msg) on the testicular morphology and cauda epididymal sperm reserves of young and adult male rats. Vet Arh. 2011; 81(4): 525-534.

28. Bairagi G., Kabra A., Mandade R. Sexual dysfunction in men with diabetes. Res J Pharm Tech. 2011; 4(5): 677-684.

29. Samala A, Pawar S, Manala S, Chada S, Nageshwar M. RP-HPLC method development and validation of tadalafil in tablet dosage form. J Chem Pharm Res. 2013; 5(4): 315-318. doi:10.52711/0974-4150.2021.00065

30. Arcaniolo D, Imbimbo C, Grillo M, Fusco F. Vardenafil for the treatment of erectile dysfunction. Clin Pract. 2012; 9(1): 39-48. doi:10.2217/CPR.11.131

31. Dadras J, Herman N, Rahimi S. The study of herbal medicines and their therapeutic effects for farm animals. Asian J Pharm Technol. 2020; 10(3): 137. doi:10.5958/2231-5713.2020.00024.0

32. Kusumawati I, Mahatmaputra S, Hadi R, et al. Aphrodisiac Activity of Ethanolic Extracts from the Fruits of Three Pepper Plants from Piperaceae Family. J Farm Dan Ilmu Kefarmasian Indones. 2021; 8(2): 194. doi:10.20473/jfiki.v8i22021.194-199

33. D’Cruz SC, Mathur PP. Effect of piperine on the epididymis of adult male rats. Asian J Androl. 2005; 7(4): 363-368. doi:10.1111/j.1745-7262.2005.00059.x

34. Munglue P, Kupittayanant S, Kupittayanant P. Effect of watermelon (Citrullus lanatus) flesh extract on sexual behavior of male rats. Chiang Mai Univ J Nat Sci. 2014; 13(1): 519-527. doi:10.12982/cmujns.2014.0054

35. Cormio L, De Siati M, Lorusso F, et al. Oral L-citrulline supplementation improves erection hardness in men with mild erectile dysfunction. Urology. 2011; 77(1): 119-122. doi:10.1016/j.urology.2010.08.028

36. Jain MS, Koradia SK. Phosphodiesterase-5 (PDE5) Inhibitors in the Treatment of Erectile Dysfunction: A Review. Asian J Pharm Res. 2023; 13(1). doi:10.52711/2231-5691.2023.00012

37. Lasker GF, Pankey EA, Kadowitz PJ. Modulation of soluble guanylate cyclase for the treatment of erectile dysfunction. Physiology. 2013; 28(4): 262-269. doi:10.1152/physiol.00001.2013

38. Kagathara V, Vyawahare N, Pujari R, Patil M, Ansari I, Bhandare A. Effect of Chlorophytum borivilianum on Fluoxetine Induced Sexual Dysfunction in Female Rats. Res J Pharmacol Pharmacodyn. 2009; 1(3): 130-133.

39. Carro-Juárez M, Alcazar C, Ballesteros-Polvo E, Villalobos-Peñalosa P. Increase of ejaculatory capacity by systemic administration of the oquichpatli (Senecio cardiophyllus) aqueous crude extract in male rats. J Ethnopharmacol. 2009; 126(3): 506-511. doi:10.1016/j.jep.2009.09.006

40. Singh S, Nair V, Gupta YK. Evaluation of the aphrodisiac activity of Tribulus terrestris Linn. in sexually sluggish male albino rats. J Pharmacol Pharmacother. 2012; 3(1): 43-47. doi:10.4103/0976-500X.92512

Received on 13.10.2023 Modified on 30.11.2023

Research J. Pharm. and Tech 2024; 17(8):3969-3975.

DOI: 10.52711/0974-360X.2024.00616

Yield of extracts and detection of components	Detected active components in the extracts
Yield of extracts and detection of components	Red chili (PRE)	Eggplant fruit (EE)	Sweet basil leaf (OBLE)	Watermelon albedo (WAE)	Red ginger (ZRE)
Yield (% b/b)	11.28%	29.3%	13.48%	24.79%	6.31%
Alkaloids	+	+	+	+	+
Flavonoids	+	+	+	+	+
Tannins	-	Undetected	-	Undetected	Undetected
Saponins	+	+	+	+	+
Terpenoids/Steroids	+	+	-	Undetected	-

NC	*SD 10*	PRE 10
PRE 30	EE 10	EE 30
OBLE 10	OBLE 30	WAE 10
WAE 30	ZRE 10	ZRE 30