EVALUATION OF APHRODISIAC ACTIVITY OF TAMILNADIA ULIGNOSA (RETZ.) BARK

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ABOUT AUHTORS
Neerugatti Dorababu, Battu Ganga Rao, Devarakonda Ramadevi

A.U. College of Pharmaceutical Sciences, Andhra University,
Visakhapatnam, Andhra Pradesh (A.P), India
*ramapathi.addepalli@gmail.com

ABSTRACT:
The study was aimed at investigate the effect of methanolic extract of Tamilnadia ulignosa (Retz.) (Rubiaceae) on general mating behaviour, libido, and adverse effects on sexually normal male albino rats.  Methanolic extract was administered orally at the dose of 100, 200, and 400 mg / kg, to different groups of male rats (n = 8) once a day for 14 days . All the doses resulted in significant increase in mount frequency, intromission frequency and anogenital sniffing when compared to normal. The methanolic extract of Tamilnadia ulignosa (Retz.) bark at higher concentration (400 mg/kg body weight) showed significant aphrodisiac activity on male Wister albino rats as evidenced by an increase in number of mounts and mating performance. Thus, in experimental rats, the results of the present study suggest that the methanolic extracts of Tamilnadia ulignosa (Retz.) exert significant aphrodisiac activity

REFERENCE ID: PHARMATUTOR-ART-2481

PharmaTutor (Print-ISSN: 2394 - 6679; e-ISSN: 2347 - 7881)

Volume 5, Issue 4

Received On: 12/12/2016; Accepted On: 23/12/2016; Published On: 01/04/2017

How to cite this article:Neerugatti D, Battu GR, Devarakonda R;Evaluation of aphrodisiac activity of tamilnadia ulignosa (retz.) bark; PharmaTutor; 2017; 5(4);55-65

INTRODUCTION:
The term aphrodisiac is derived from Greek mythology, where Aphrodite was the goddess of love and beauty [1] . The Greek word ‘aphrodisia’ means sexual pleasure [2]. The modern definition of aphrodisiac can vary, but is generally regarded as a substance that increases sexual desire (i.e. libido) and/or sexual pleasure. The definition has been extended to include those substances which enhance sexual performance or aid in the proper functioning of the male and female sex organs [3,4]. Substances include foods, beverages, vitamins, minerals, and other natural and synthetic chemicals[5] . There are numerous reports of aphrodisiac activity attributed to plants [6,7,8,9,10,11,12], isolated constituents. [13]

MATERIALS AND METHODS
Preliminary Phytochemical studies The powder of dried leaves of Tamilnadia ulignosa (Retz.) (family Rubiaceae) was subjected to continuous soxhlet extraction with various organic solvent such as methanol. The 1:3 ratio for drug to solvent was maintained for each solvent for successive solvent extraction method. After concentration and drying of each extract in vacuum desicator, identification of phytoconstituents was carried out using thin layer chromatography method by different detecting reagents. 11 The all extracts  were then subjected to qualitative chemical tests for confirmation of phytoconstituents of interest.

Preparation of extracts  The leaves of Tamilnadia ulignosa (Retz.) were collected from Tirumala hills, Chitoor, Andhra Pradesh, India and authenticated by Dr. Madhava shetty, taxonomist, Department of Botany, Sri Venkateswara University, Andhra Pradesh. Shade dried leaves of Tamilnadia ulignosa (Retz.)  were powdered and separately extracted in a Soxhlet apparatus for 6 hrs with methanol and then concentrated under vacuum at temperature of 45oc by using rotary evaporator (Buchi), dried completely, weighed and stored in desiccators.

Requirements Sildenafil citrate [Shilpa antibiotics, Raichur, India.] Progesterone capsules (100 mg) [Alembic limited, Vadodara, India] Oestradiol valerate injection (10mg/ml) [German Remedies, Mumbai.] Olive oil [s.d.fine Chem Ltd. Mumbai ] Ethyl Alcohol [The Ugar Sugar Works Ltd. Ugar Khurd, Belgaum] REMI research centrifuge (R-24) Borosil Soxhlet extractor Solvent evaporator. Research microscope (Metzer) Afcoset Digital balance(E-R-180 A) Mini Lyotrap Dryer (LTE Scientific Ltd, Great Britain) were purchased from Sigma chemicals, USA.  All chemicals used in the study were of analytical grade.

 

Animals Albino rats (National Institute of Nutrition, Hyderabad, India) of either sex weighing 200-250gm were used in the studies. The animals were maintained under standard laboratory conditions at an ambient temperature of 23±2±C having 50+-5% relative humidity with 12h light and dark cycle. The use and care of the animals in the experimental protocol has been approved by the local Institutional Animal Ethics Committee (Regd. No. 516/01/A/CPCSEA) following the guidelines of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Government of India.

Evaluation for aphrodisiac activity[14,15] Adult, healthy male Albino rats weighing between (150-200 g) were divided into eight groups, each consisting of five rats. Group-A:  Received distilled water (normal control) p.o. Group-B:  received dose of Sildenafil citrate (0.7 mg/kg) p.o. Group-C:  received low dose of Tamilnadia Ulignosa (Retz.) (100mg/kg) p.o. Group-D: received medium dose of  Tamilnadia ulignosa (Retz.) (200mg/kg) p.o. Group-E:  received high dose of Tamilnadia ulignosa (Retz.) (400mg/kg) p.o. All the treatments were given continuously for 14 days. On the 14th day, after treatment, all animals were allowed for mating and were recorded for evaluating aphrodisiac activity. All the animals showed quick mount latency, increased number of mounts, delayed intromission latency and increased number of intromissions in comparison to normal animals. All the animals showed delayed inter intromission interval and able to attain more intromissions and more ejaculations in comparison to normal animals (Table).  

Effect on Stress modulated sexual behavior in male rats Male Albino rats weighing between (250-300 g) were divided into following leaven groups, each group consisting of five rats. Group-A: normal control (receives distilled water 3ml/kg) (without stress) p.o. Group B: stress control (receives distilled water 3ml/kg) p.o. Group-C:  received low dose of Tamilnadia ulignosa (Retz.) (100mg/kg) p.o. Group-D: received medium dose of  Tamilnadia ulignosa (Retz.) (200mg/kg) p.o. Group-E:  received high dose of Tamilnadia ulignosa (Retz.) (400mg/kg) p.o. Group-F:  received dose of Testosterone 0.05mg  per rat i.m. All the treatments were given for 28 consecutive days before Immobilization stress. [16]

Induction of Immobilization stress  The animals were subjected to IMB stress by Plexiglas cylinder (5 cm diameter and 16 cm large) for 6 h a day during light period started from 10 am each day for 28 consecutive days. Water and food were withdrawn during stress period.

Sexual accessory Organ to body weight ratio
Body weight of each animal was measured on day 1 before the IMB stress and on day 28 after IMB stress and drug treatment. The percentage change in body weight was calculated. Animals were sacrificed on day 28 by euthanasia with ether, accessory sexual organs viz, testis, vasdeferens, prostate glands, seminal vesicles, epididymis and adrenal glands were isolated and weighed in wet condition to measure organ to body weight ratio.

Sperm count
1.    Spermatozoa were collected by flushing the vas deferens and epididymis in 2.0ml of normal saline. Draw the semen in the WBC pipette up to 0.5 mark. Draw in 4% sodium bicarbonate in 1% phenol solution up to the mark 11, making a dilution of 1 in 20. Count the sperm under high power in the four WBC squares.
    Calculation: Number of sperms in 1 cu.mm of sample = N x 10/4 x 20
                          Number of sperms in 1 ml (i.e.1 cu.cm) of sample:
                                              = N x 50 x 1000(as 1 cu.cm=1000cu.mm)
                                              = N x 50,000

Where N is the total sperm count observed in outer four square of WBC chamber

Sperm motility Place a drop of semen on the cover slip and invert it on a rim of plasticine on the cavity slide. Examine under high power objective and find out the percentage of immobile to mobile sperms.

Histology of testis
Two-left testis of each group was excised and rinsed in 0.9% saline blotted dry of saline and excess blood. They are fixed in 12 % formalin for 24 hrs. The tissues, after fixation, were washed in water to remove excess fixative. Washed   tissues were dehydrated through a graded series of ethyl alcohol, cleared with xylene and embedded in paraffin wax. Sections were cut at 3 μm with microtone blade, and mounted on clean glass slide. The sections were routinely stained with haemotoxyllin and eosin. The stained slides were observed (400 X) in research microscope and photographed.

RESULTS
Table 1:Qualitative chemical examination of different extract of seeds of Tamilnadia ulignosa (RETZ.) bark (TU)

S.No

Phytoconstituents

TU

1

Alkaloid

-

2

Carbohydrate

+

3

Phytosterol

+

4

Protein & Amino acid

-

5

Saponin Glycoside/ glycoside

+

6

Fixed oil / fat

+

7

Gum/ mucilage

+

8

Flavonoids

+

9

Volatile oil

-

10

Amino acids

+

11

Tannin

+

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