Formulation and evaluation of Antimicrobial activity of Elaeocarpus Ganitrus leaves

Elaeocarpus ganitrus, also known as Rudraksha in Sanskrit or Rudraki in Hindi, is one of the most significant herbal plants used in indigenous medical systems like Ayurveda, Siddha, and Unani. The E. ganitrus plant’s ripe fruit is made up of a hard, rocky endocarp known as a bead, nut, or Rudraksha. The rudraksha plant is the most well-known plant with spiritual and therapeutic uses ¹. The Rudraksha plant has historically been used to treat conditions including stress, anxiety, palpitations, depression, nerve pain, migraines, sleeplessness, acne, ringworm, epilepsy, asthma, poor concentration, arthritis, hypertension, and liver problems. Analgesic, antidepressant, antidiabetic, anticonvulsant, cardio-stimulator, hydrochloric, anti-aging, antiseptic, antihypertensive, hypnotic, antiepileptic, tranquilizing, anti-inflammatory, thermogenic, sedative, smooth muscle relaxant, and hydro choleretic are just a few of the pharmacological effects of the Rudraksha plant.²

Botanical Description: It is a large evergreen tree with large leaves. Its height ranges from 50–200 feet. Leaves are large and shining green on the sun facing side and dull stringy on earth facing side. Flowers become visible in the month of April May and are white or yellow in colour^3,4. Fruits start appearing in June and ripen near October. Ripe fruit is fleshy and has a seed with blue shell. Inner part or bead lying in the seed is called Rudraksha³.

Morphological & Macroscopically Description 2,4,5,6

The morphological characters of plant are shown below as follow as,

Leaves- Simple, glabrous, oblong-lanceolate, sub-entire or irregularly crenate, acute or acuminate

Flower- White or yellow colored, in dense racemes and mostly from axils of fallen leaves, fringed petals, anthers are linear, appear in April-may

Fruits- Round or oval, small, violet or blue colored and

Taste: Acidic  Endocarp- Stony endocarp is hard, globular, strongly tubercule, marked with 5 to longitudinal ridges, rarely 1 to 4,

Color :reddish brown

Acne  vulgaris  is an  extremely  common disorder  of the skin  (pilocebaceous  unit)  that  affects  virtually  all individuals  at  least once  during  life.  The  incidence  of acne peaks of teenage,  but substantial  numbers of  men and women between 20-30 years of age are also affected. by the disorder.⁷  herbal as well as synthetic ingredients are reported to have remarkable beneficial effect on acne vulgarisThey may  have  different  mechanisms  like, (a)  Control  sebum  secretion,  (b)  Antibiotics  which inhibit  acne,  the  main  causative organism  of  acne,  (c)  Keratolytic  which  removes  the keratin layer  and prevents  the trapping  of sebum under the  skin,  (d)  Anti-inflammatory  which  prevents  the worsening  of  the  condition  due  to  inflammation  or redness etc. Numbers of formulations are available in the market  with  a  variety  of  active  pharmaceutical ingredients for the treatment of acne.^8,9,10  Topical preparations such as gels, ointments, lotions and creams are the important drug delivery systems due to its convenience in delivering drug to a localized area of the skin.  Gel  is  one  of the  semi-solid  topical  preparations providing quick onset of activity, long-term efficacy and

high patient satisfaction.

MATERIALS AND METHODS

Plant Materials and Preparation of Extract: Rudraksh leaves leaves were collected from Himalaya, cleaned from foreign material, washed with distilled water, dried in the shade for 72 hours, coarsely grinded, weighed, and stored in airtight jars. One liter of ethanol (95% v/v) was added to 250 g of powdered Rudraksh leaves for 3 to 4 days. The mixture was stirred with a sterile glass rod after 12 h and was filtered with Whatman filter paper No. 1. In a rotary evaporator, the solvent was removed under reduced pressure at a temperature of less than 50 °C, leaving a dark green residue stored in the airtight glass jars. The extract’s weight was recorded and the percentage yield was 10.6%. Carbopol 940, Propylene glycol, Sodium hydroxide and Methyl paraben .

The phytochemical screening of the  alcoholic extracts: 

The leaf of  rudraksh leaves was carried  out for the presence  alkaloids,  cardiac  glucosides,  flavonoids phenols, resins,  saponins, tannins,  terpenes and steroids using  standard  phytochemical  methods.  The phytochemical screening of the ethanolic extracts of the plant was carried out  in order  to elucidate  the chemical constituents  (bioactive  agents)  responsible  for  their antimicrobial and therapeutic activities

Preparation of Gel Formulations :

Distilled water was added to the Carbopol 940 and mixed mechanically  by  high-speed  mixer.  To  this  mixture, sodium hydroxide 10% was added vigorously.  In water bath  with  a  temperature  not  exceeding  50  °C,  the extracts in  a  concentration of  1,  2, and 3 g were added to prepare three formulations, Gel-A (1%  w/w),  Gel-B  (2%  w/w)  and  Gel-C (3%  w/w), respectively.  Separately  dissolved  methyl  paraben  in propylene  glycol  were  also  added  to  this  gel.  The remaining quantity of purified water was added,

Evaluation of Gel Formulations

Physical appearance

The  gel formulations  were  evaluated for  their  physical parameters  like  color,  odor,  consistency,  transparency, and homogeneity.

Spreadability of Gel Formulations

A glass slide with standard dimensions was used, where 0.5 g of the gel was placed in a circle 1 cm in diameter on  the glass  slide, over  which  another  glass slide  was placed. A weight of 125 g was set for 5 min so that the gel  was sandwiched  between  the  two  slides  to  form  a thin layer. Then, the weight was removed and the extra gel was removed. Then  the slides  were adjusted  so that the upper  slide was  fixed  with a  weight of  about 20 g. The time was noted fonoted the slides to separate from each other.

The  spreadability  was  recorded  using  the

following formula.

S = M / T

Where:

S – Spreadability in grams/seconds;

M – Mass in grams;

T – Time in seconds.

Viscosity.

A Brookfield DV-E viscometer (RVDVE) was used to determine the viscosity of the gels. Spindle No. 07 was inserted  in each  formulation and  was sheared at 3.3, 9.9, and 16.5  g at 24 ± 1  °C.

The  gel formulations were prepared in distilled water

PH Determination

The pH of the gels was detected with a digital pH meter. An  amount  of 0.5  g  of  gel was  dissolved  in 50  ml  of distilled  water  and  stored  for  two  hours.  Each formulation’s  pH  was  measured  in  triplicate  and  the average values were taken.

Antibacterial Activity of Gel Formulations

Each  formulation  was  assessed  for  its  antimicrobial effects  against  the  microorganisms  on  a  nutrient  agar using a  suitable diffusion  method. About  0.2 ml  of the bacterial test strain was inoculated  over  a nutrient agar plate with a sterile cotton swab and was allowed to dry. With the help of a cork borer, 6 mm diameter wells were created. Half a milliliter of the Rudraksh leaves extract was introduced into the wells. The  plates  were  placed  at  room  temperature  for  about one hour. Then the plates were placed in an incubator at 37 °C  for 24  hours.  Then, the  zone  of inhibition  was checked  and  recorded.  Clindamycin  was  used  as standard

Acne Healing Activity of Gel Formulations

Adults aged from 17 to 22-year-old were divided into three groups, having 4 adults each. Group A, B, and C received Gel-A containing 1% w/w of the Rudraksh   leaves  extract, Gel-B containing 2% w/w of the leaves extract   and Gel-C containing 3% w/w of the extract. No other medicine was given to the adults during the entire study. The study was evaluated for 15 days

RESULT AND COCLUSION:

This study evaluated the anti-acne potential of herbal gels. Three different concentrations of an Rudraksh leaves extract were used to prepare gel formulations with Carbopol 940. The formulations were evaluated for the physical parameters like the pH, viscosity, and spreadability.

A pharmacological evaluation:, like a skin irritation test, revealed that the herbal gels were safe to apply on the skin. The antibacterial activity of these gels against acne bacteria was also tested and confirmed. An anti-acne study was carried out to show that the herbal gels can heal the acne without severe adverse effects.

Phytochemical Analysis of gel

Many phytochemicals were found in the ethanolic extract. Different tests were performed according to the standard methods to check for the presence of phytoconstituents such as alkaloids, flavonoids, tannins, reducing sugars, saponins, triterpenes and glycosides in the ethanolic extract of the Rudraksh leaves The observations were recorded .

Phytochemical Constituents of Ethanolic Extract of Rudraksh leaves.

Evaluation of Gel Formulations

All the formulations were green a/c to percentage

The spreadability :.

The bioavailability efficiency of a gel formulation also depends on its spreading value. pH: alkaline  which was compatible with normal skin physiology. The results of the viscosity are also shown below  in table.

Skin Irritation Test

All the gel formulations were found to be safe while being applied on the skin and there was no irritation or sensitivity to the skin.

Antibacterial Activity of Gel Formulations

The antibacterial activity showed Table  that the zone of inhibition increased with an increase in the concentration of the herbal extract. It indicates that the Rudraksh leaf extract possesses an antibacterial activity, helps maintain a sterile acne area, and promotes the acne healing process. Gel-C was found to be more effective in the acne healing when compared to other herbal gels. These gels showed better activity against acne bacteria.