Global J Res. Med. Plants & Indigen. Med.

| Volume 1, Issue 9 | September 2012 | 381–390

ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal

Research article

APHRODISIAC STUDIES OF DIHERBAL MIXTURE OF

ZANTHOXYLUM LEPRIEURII GUILL. & PERR. AND
PIPER GUINEENSE SCHUMACH. & THONN. ON MALE WISTAR RATS

Kpomah E D1*, Uwakwe A A2, Abbey B W3

1, 2, 3
Department of Biochemistry, University of Portharcourt, Rivers State, Nigeria.
*
Corresponding Author: Ph: +2348034173608; E-mail: [email protected]

Received: 09/08/2012; Revised: 24/08/2012; Accepted: 29/08/2012

ABSTRACT

The effects of 50, 100 and 150 mg/kg body weight of ethanol extract of diherbal mixture of
Zanthoxylum leprieurii and Piper guineense on aphrodisiac potentials and hormonal levels was
investigated. A total of twenty healthy, sexually experienced Albino male rats (Rattus norvergicus),
weighing between 190–230g were randomly divided into four groups (A–D) of five rats each, group
A were orally administered once daily with 1 ml of distilled water (vehicle), while groups B, C and
D received 50, 100 and 150 mg/kg body weight of the extract in 1ml of the vehicle. After 21 days of
treatment the animals were studied for mating behaviour and hormonal assay, the mating behaviour
monitoring showed that mount latency, intromission latency and post ejaculatory intervals were
significantly decreased (P < 0.05), mount frequency, intromission frequency, ejaculatory frequency
and ejaculatory latency were all significantly increased (P < 0.05), however, only the 150 mg/kg
body weight dosage produced significant increase in copulatory efficiency. Hormonal assay showed
that all doses produced significant increase (P < 0.05) in testosterone, luteinizing hormone and
follicle stimulating hormone, similarly all doses produced significant increase (P < 0.05) in
progesterone except 50 mg/kg, prolactin was decreased significantly (P < 0.05) by all dose level. The
effects of these extracts on the mating and hormonal studies were dose dependent and the findings
validated the acclaimed use of this herbal product as an aphrodisiac in men.

KEYWORDS: Aphrodisiac, Diherbal mixture, Zanthoxylum leprieurii and Piper guineense

Cite this Article

Kpomah E D, Uwakwe A A, Abbey B W (2012), Aphrodisiac studies of diherbal mixture of
Zanthoxylum leprieurii Guill. & Perr. and Piper guineense Schumach. & Thonn. on Male wistar rats,
Global J Res. Med. Plants & Indigen. Med., Volume 1(9), 381–390

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Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 9 | September 2012 | 381–390

INTRODUCTION sufferers often shy away from these treatment

options and these could be attributed to its
The incidence of male sexual dysfunction is sensitivity and social stigma attached to male
on the increase globally (Kaiser, 1999). It is sexual dysfunction in the African context (Lim
reported as having a prevalent rate of 10% et al., 2005), these treatment options are
across all ages (Lauman et al., 1999), and equally very expensive, not easily accessible to
because sexual dysfunction is an inevitable the poor and rural dwellers and are often
process of aging, the prevalence is over 50% in associated with some serious side effects,
men between 50 and 70 years of age (Rendell consequently medicinal plants with marked
et al., 1999). pharmacological activities are readily available
all year round, cheap and accessible and often
Male sexual dysfunction is classified as a with minimal side effects (Tilburt and
disorder of desire that is persistent absence of Kaptchuk, 2008; Ufelle et al., 2011; Yadav and
sexual fantasy and desire for sexual activity Agarwala, 2011), and are being explored
(Kandeel et al., 2001), erectile dysfunction that globally as panacea. All through history many
is persistent inability to develop and maintain a preparation from plants have been used and
penile erection that is for intercourse and reputed to have sex invigorating (aphrodisiac)
ejaculation in 50% or more of attempts properties and these include Yohimbine,
(Kandeel et al., 2001), disorder of ejaculation Gingseng, Massularia acuminata, Montanoa
that is problem with expulsion of semen at the tomentosa (Ang et al., 1997; Baljinder et al.,
climax of the sexual act, disorder of orgasm 2010; Yakubu and Akanji, 2011).
that is delay in or absence of orgasm after a
normal sexual excitement phase during sexual The composite extract of the bark of
activity (Rosen and Lieblum, 1995), failure to Zanthoxylum leprieurii Guill. & Perr. and the
detumescence that is prolonged (> 4 hours seeds of Piper guineense Schumach. & Thonn.
duration) and extreme painful erection are two of such sex tonics that is largely used
unaccompanied by sexual desire (Kandeel et by the people of Niger Delta region of Nigeria,
al., 2001). but with no scientific footing. Z. leprieurii
belong to the family Rutaceae and locally
The causes of male sexual dysfunction called Prickly ash or toothache tree, it is an
include factors like psychological disorders aromatic, spiny, thicket forming deciduous
(performance anxiety, strained relationships, shrub or tree. The alternate branches are armed
depression, stress, guilt, fear of sexual failure), with strong brown prickles about 2–3 cm long,
hormonal condition (androgen deficiency, cone shaped with a broad base and found
hyperprolactinemia), chronic medical irregularly throughout the tree (Todd, 2008).
conditions (diabetes, hypertension), penile Ethnomedically, it is used in the treatment and
disease (priapism, smooth muscle dysfunction), management of muscle spasm, varicose vein,
neurological disorder (Parkinson disease, raynauld disease, arthritis, rheumatism,
stroke, cerebral trauma, alzheimer’s disease, neuraligia, flu, fever, toothache and gum
spinal cord or nerve injury), drug side effect diseases (Tilloston, 2011). P. guineense
(antihypertensive, psychiatric medications, commonly called climbing pepper is a slender
antiulcer and antidepressants), lifestyle (chronic climber up to 12 m high with prominent nodes
alcohol abuse, cigarette smoking), aging and and clasping roots, the leaves are elliptic in
systemic disease (cardiac, hepatic, renal, shape about 15 cm long and 7 cm broad, the
pulmonary, metabolic), post organ transplant flowers are small, borne on common stalk as
(Kandeel et al., 2001; Guay et al., 2003; cluster opposite to the leaves. The fruits are red
Yakubu et al., 2007). and turns black when dry (Iwu, 1988).
Traditionally the seeds are used as spices and
Despite the medical advancements in
as sex invigorator in men.
treatment and treatment facilities for male
sexual dysfunction (Lim et al., 2005), most

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Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 9 | September 2012 | 381–390

The present study was therefore undertaken dark cycle with free access to grower’s mash
to evaluate the aphrodisiac property of the and water.
composite ethanol extract of the stem bark of
Zanthoxylum leprieurii and the seeds of Piper Experimental Design
guineense using a dose level of 50, 100 and
The twenty male rats were randomly
150 mg/kg body weight with a view to validate
divided into four groups (A–D), consisting of
the acclaimed use of this sexual invigorator.
five rats each. Group A were orally
MATERIALS AND METHODS administered once daily with 1 ml of distilled
water (vehicle), groups B, C, and D were orally
The plant samples were bought in a local administered with 50, 100 and 150 mg/kg body
herb market in Warri, Delta State, Nigeria. weight of the extract in 1 ml of the vehicle for
Both the species were identified and confirmed 21 days.
at the Herbarium of the Department of Plant
Science and Biotechnology of the University of Mating Behaviour Test Procedure
Portharcourt, Rivers State. The investigation was carried out on the
Preparation of Plant Extract 22nd day after the commencement of extract
administration by adopting the methods
The bark Z. leprieurii and the seeds of P. described by (Gauthaman et al., 2002; Yakubu
guineense were thoroughly washed with and Akanji, 2011). The investigation was
distilled water to remove debris and conducted between 19 and 22 h in the same
contaminants, they were then dried in an oven laboratory and under a dim light. The female
at 40℃ until a constant weight was reached, rats were artificially brought to oestrus by
and then pulverized using an electric blender administering estradiol benzoate 10 µg/100 g
(Blender, 462 Nakai Japan). 200 g of the body weight orally 48 h prior to mating and
powdered mixture (i.e 100 g each of Z. progesterone injected subcutaneously at a dose
leprieurii and P. guineense) was extracted in of 0.5 mg/100 g 6 h prior to the mating
600 ml of absolute ethanol for 24 h at room (Szectman et al., 1991; Yakubu et al., 2005).
temperature with constant shaking using a flask The artificially warmed female was introduced
shaker (Model, Denly A - 500). The extract to the cage of the male. The male and female
was filtered with Whatman No. 1 filter paper were observed from the cage side for
and the resulting filtrate was evaporated to proceptive, precopulatory and copulatory
dryness using a rotatory evaporator at 40℃ to behaviours. The occurrence of events and
give 5.74 g, the resultant concentrate was phases of mating was monitored for 30 min
reconstituted in distilled water to give the observatory period and the following male
required doses used in the study. sexual behavior indices recorded/calculated.
Mount latency: the time interval between the
Experimental Animals introduction of the female to the first mount by
the male; Mount frequency: the number of
A total of twenty healthy sexually
times the male assumed copulatory position but
experienced Albino male rats (Rattus
failed to achieve intromission; Intromission
norvergicus), 2.5–3 months old, weighing
latency: the time interval from the introduction
between 190–230 g and thirty female rats
of the female until the first intromission of the
between 2–2.5 months old and weighing
female (vaginal penetration) by the male;
between 150–180 g were obtained from the
Intromission frequency: the number of
animal house unit of the Department of
intromissions (vaginal penetration) made by the
Biochemistry, University of Portharcourt,
male from the first time of introduction of the
Rivers State. The animals were kept in a clean
female; Ejaculatory latency: the time interval
cage and housed in a well ventilated room at
between the first intromission and ejaculation,
temperature 28–30℃ under natural light and
it is usually characterized by longer, deeper

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Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 9 | September 2012 | 381–390

pelvic thrusting and slow dismount, followed means were analysed using one way analysis of
by a period of reduced activity; Ejaculation variance (ANOVA) followed by Posthoc
frequency: The number of times there was (Turkey). P < 0.05 was regarded as significant.
expulsion of semen by the males after vaginal The Statistical Package for Social Sciences
penetration characterized by rhythmic (SPSS) Computer software version 16 was used
contraction of the posterior abdomen; Post for data analysis.
ejaculatory interval (PEI): the time interval
from ejaculation to intromission of the next RESULTS
mating series; Copulatory efficiency:
Effects of the Composite Mixture of Z.
number(s) of intromission divide by number(s)
leprieurii and P. guineense on Mating
of mount multiplied by 100 %. (Yakubu et al.,
Behaviour
2005, and Yakubu and Akanji, 2011).
Upon the introduction of the female to the
Method of Collection and Handling of
male cage, the male responded with immediate
Serum advances towards the female and displayed
At the end of the treatment period, the precopulatory behaviours such as chasing,
animals were anaesthetized in a chloroform anogenital sniffing which eventually
chamber and blood samples collected from the culminated into mounting, the female also
jugular vein into sample bottles, the blood displayed receptivity that allowed mating to
samples were allowed to clot for 10 min at occur. The ethanol extract of the diherbal
room temperature and subsequently centrifuged mixture of Z. leprieurii and P. guineense after
to obtain serum for hormonal assay. 21 days of treatment at a dosage of 50, 100 and
150 mg/kg body weight was able to influence
Assay Kits significantly (P < 0.05) on the mount latency,
intromission latency, mount frequency,
Testosterone, follicle stimulating hormone intromission frequency, ejaculatory latency,
(FSH), luteinizing hormone, prolactin and ejaculatory frequency and post ejaculatory
progesterone radioimmunoassay test kits are interval, however, only the 150 mg/kg body
products of BYK-Sangtic Diagnostica, GmbH weight of the extract had significant effect
and Co. KG, while estradiol benzoate and (P < 0.05) on the copulatory efficiency (Table
progesterone are products of Sigma Chemicals, 1.0).
St. Louis, USA and Shalina Laboratories,
Mumbai, India, respectively. Effects of the Composite Mixture of Z.
leprieurii and P. guineense on Hormonal
Hormonal Assay Levels.
Serum samples were assayed for the The ethanol extract of the composite
following hormones testosterone, follicle mixture of Z. leprieurii and P. guineense after
stimulating hormone, luteinizing hormone, 21 days of treatment at 50, 100 and 150 mg/kg
prolactin and progesterone by using the body weight dose was able to significantly
procedure described by BYK-Sangtic increase (P < 0.05) the levels of testosterone,
Diagnostica. This was based on the principle of luteinizing hormone and follicle stimulating
radioimmunoassay of competitive binding hormone, however, only 100 and 150 mg/kg
between the sample serum and the standards for body weight was able to significantly
a constant amount of the antisera. (Tietz, 1995). (P < 0.05) increase progesterone. All doses of
the extract studied were also able to
Statistical Analysis significantly decrease (P < 0.05) the level of
prolactin (Table 2.0).
The results were expressed as the mean of
five replicates ± standard deviation (S.D),

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Table 1.0 Effect of 50, 100 and 150 mg/kg Body Weight of Ethanol Extract of Composite of
Zanthoxylum leprieurii and Piper guineense on Mating Behaviour Parameters.

C D
MATING B
S/N A CONTROL 100 mg/kg 150 mg/kg
PARAMETERS 50 mg/kg B.W
B.W B.W
Mount Latency
1. 813.00 ± 4.95 a 746.00 ± 6.67 b 610.00 ± 7.21 c 493.00 ± 9.76 d
(Seconds)
Intromission Latency
2. 820.00 ± 7.91 a 750.00 ± 6.63 b 618.00 ± 5.70 c 499.00 ± 8.63 d
(Seconds)
3. Mount Frequency 5.20 ± 0.84 a 10.40 ± 2.07 b 14.20 ± 1.92 c 18.80 ± 1.48 d
4. Intromission frequency 3.00 ± 0.71 a 6.80 ± 1.64 b 9.60 ± 1.52 c 13.00 ± 1.58 d
Ejaculatory Latency
5. 234.00 ± 6.52 a 274.00 ± 9.19 b 310.00 ± 11.81c 409.00 ± 8.54 d
(Seconds)
6. Ejaculatory Frequency 1.00 ± 0.00 a 2.00 ± 0.00 b 3.20 ± 0.45 c 4.20 ± 0.84 d
Post Ejaculatory
7. 400.00 ± 7.91 a 353.40 ± 8.44 b 308.60 ± 6.84 c 241.20 ± 7.01 d
Interval (Seconds)
Copulatory Efficiency
8. 57.33 ± 7.23 a 65.01 ± 3.70 a 67.49 ± 2.46 a 69.00 ± 360 b
(%)
Values are means of five replicates ± standard deviation, values with different superscript letters b, c and d are
significantly different (P < 0.05) from the control ‘a’. BW: body weight.

Table 2.0 Effect of 50, 100 and 150 mg/kg Body Weight of Ethanol Extract of Composite of Z.
leprieurii and P. guineense on Hormonal Levels.

B C D
A
S/N 50 mg/kg 100 mg/kg 150 mg/kg
HORMONE CONTROL
B.W B.W B.W
1. Testosterone (nmol/L) 3.10 ± 0.16 a 4.22 ± 0.19 b 7.94 ± 0.21 c 10.62 ± 0.22 d
2. Luteinizing hormone (IU/L) 2.86 ± 0.19 a 3.64 ± 0.38 b 4.98 ± 0.29 c 5.30 ± 0.31 a
3. Follicle stimulating hormone 2.92 ± 0.23 a 4.12 ± 0.33 b 4.98 ± 0.15 c 6.08 ± 0.28 d
(IU/L)
4. Progesterone (nmol/L) 0.98 ± 0.24 a 1.34 ± 0.24 a 1.44 ± 0.18 b 1.64 ± 0.18 c
5. Prolactin (ng/ml) 6.82 ± 0.37 a 6.14 ± 0.32 b 5.60 ± 0.46 c 5.04 ± 0.19 d
Values are means of five replicates ± standard deviation, values with different superscript letters b, c and d are
significantly different (P<0.05) from the control ‘a’. B.W: body weight.

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DISCUSSION
Ejaculatory latency and ejaculatory
Aphrodisiacs can be defined as substances frequency are pointers of enhanced copulatory
which are ingested, applied topically, performance, the extract was also able to
smoked/snorted or otherwise delivered into the significantly increase ejaculatory latency and
body to induce sexual arousal, heighten sexual ejaculatory frequency (P < 0.05). Ejaculatory
experience and to improve sexual performance. latency also implies prolonged coitus duration
History had it that many preparations from which translates into increased staying power,
plants and animals have been used for this strength and vigour thus validating its
purpose e.g. Yohimbine, the mandrake plant, aphrodisiac properties just other aphrodisiac
Gingseng, ground rhinocerous horn, sheep and plants that have been investigated e.g. Vanda
bull testicle and the Spanish fly (Ang et tsellata (Suresh et al., 2000), Dactylorhiza
al.,1997; Baljinder et al., 2010). This present hatagirea (Thakur and Dixit, 2007) and
study was therefore designed to investigate the Mondia whitel (Watcho et al., 2007).
aphrodisiac properties of the composite mixture
of Zanthoxylum leprieurii and Piper guineense. Post ejaculatory interval is a positive
marker of sexual potency, libido and a fast pace
Mount latency and intromission latency are of recovery from exhaustion after the first
indices of sexual motivation and there is an series of mating, all doses of the extract
inverse relationship between mount latency, significantly decreased the post ejaculatory
intromission latency and sexual motivation interval (P < 0.05) and compared favourably
(Yakubu and Akanji, 2011). The extract was with other aphrodisiac plants that have been
able to significantly decrease (P < 0.05) the studied e.g. Chlorophytum boriviliaunum
mount latency and intromission latency and this (Thakar and Dixit, 2006) and Syzygium
might imply stimulation of sexual appetite and aromaticum (Tajuddin et al., 2004).
arousal, thus lending credence to the sexual
improving effect of the extract under study just Copulatory efficiency is an indication that
like other aphrodisiac plants that have been the copulatory action of the male was well
studied e.g. Lepidium myenii (Cicero et al., enhanced with well coordinated pelvic
2001), Montanoa tomentosa (Carro-Juarez et thrusting and this further indicates sustained
al., 2004) and Microdemis kenyana (Zamble et increase in interest, focus, agility and stamina
al., 2008). in the sexual act. Only the 150 mg/kg body
weight of the extract was able to increase the
Mount and intromission frequencies are copulatory efficiency significantly (P < 0.05).
indicators of libido, sexual vigour, strength, Thus lending credence its aphrodisiac potency
power and energy (Yakubu et al., 2005). The which compares to other aphrodisiac plants that
extract was able to significantly increase have been studied e.g. Alpinia calcarata
(P < 0.05) the Mount and intromission (Ratnasooriya and Tayakody, 2006) and
frequencies, thus supporting the sexual Withania somnifera (Illayperuma et al., 2002).
improving effect of the extract as in the case of
other aphrodisiac plants that have been The precopulatory and copulatory
investigated e.g. Triholepis glaberrima behaviours of the extract treated rats showed
(Padashetty and Mishra, 2007), Mucuna that the rats were extremely aroused and the
pruriens (Amin et al., 1994) and Terminalia effects of the extract on the mating behaviour
catappa (Ratnasooriya and Dharmasiri, 2000). were dose dependent with the 150 mg/kg body
Increased intromission frequency is also a weight been more potent.
function of erection efficiency, penile
orientation and the ease by which ejaculatory Sexual behaviour and erection are largely
reflexes are activated (Agmo, 1997; Yakubu dependent on androgen which may act through
and Akanji, 2011). central and peripheral mechanism (Mill et al.,

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Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 9 | September 2012 | 381–390

1996; Yakubu et al., 2011). All doses of the et al., 2006), all doses of the extract was able to
extract significantly increased (P < 0.05) decrease significantly the levels prolactin in a
testosterone, luteinizing hormone and follicle manner that was dose dependent.
stimulating hormone. Luteinizing hormone and
follicle stimulating hormone are produced by The observed improvement in mating
the anterior pituitary lobe and are needed for behaviour seen between the control and the
maintaining testosterone levels, hence an extract treated groups may be due to plant
increase in luteinizing hormone and follicle chemicals present in the extract (alkaloid,
stimulating hormone automatically triggers an saponin, tannin, flavonoid, sterols), these plant
increase in testosterone levels (Yakubu et al., chemicals are able to exert their effect through
2007). Studies have shown that testosterone elevation of androgens and gonadotropins,
supplementation helps to improve sexual vasodilation and generation of nitric oxide
function and libido (Aversa and Fabiri, 2001; (Yakubu and Akanji, 2011), which are key
Grahl et al., 2007), in addition to the intensity factors in the initiation and sustenance of
of orgasm and ejaculation (Morales, 1996). All erection, libido and other sexual factors.
doses of the extract except 50mg/kg body
weight was able to increase the progesterone CONCLUSION
level significantly (P < 0.05), optimal level of
The systemic use of the ethanol extract of
progesterone have associated with improved
Zanthoxylum leprieurii and Piper guineense
sexual function (Andersen and Tufik, 2006;
have marked enhancement on mating behaviour
Andersen et al., 2007). High levels of prolactin
parameters and the sex hormones of male rats,
in men (hyperprolactinemia) have been
thus corroborating the acclaimed use of this
associated with hypogonadism, decreased
product as an aphrodisiac by the people of the
sperm count and motility, erectile dysfunction
Niger Delta region of Nigeria.
and decreased libido (Kruger et al., 2003; Paick

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Source of Support: Nil Conflict of Interest: None Declared

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