Preparation and Evaluation Antioxidant Activity of Mixed Herbal Hair Oil Formulation

About Authors: Akram Ahmad⁽¹⁾*,Vikas Kumar⁽¹⁾ ,G.P.Mohanta⁽²⁾ and Hasna Ali⁽²⁾.
1-Christian school of pharmacy, FHMSIAMS (SHIATS) Allahabad (UP)-211007.
2-Department of pharmacy practice,Annamalai University,Chidambaram (T.N)-608002.

Abstract:
Antioxidants are helpful in increasing the blood circulation and thus help in hair growth as well as in the treatment a lot of diseases. Alopecia is a dermatological disorder with psychosocial implications on patients with hair loss. The Antioxidant property of plant and oil can be  utilise in hair fall .The objective of present study involves preparation of herbal hair oil using amla, brahmi, tridex, neem, shikakai and its evaluation for increase in hair growth activity. The oils with two different drug concentrations 5% & 10% were prepared using direct boiling, paste and cloth method. For determination of antioxidant activity DPPH method is used. Herbal hair oil formulation showing moderate antioxidant activity when compared with standard.it can be used as herbal hair oil for hair growth and other Free radical damage within cells has been linked to a range of disorders including cancer, arthritis, atherosclerosis, Alzheimer's disease, and diabetes.

1-Introduction:
Herbal formulations have always attracted considerable attention because of their good activity and comparatively lesser or nil side effects as compared to synthetic drugs. Alopecia is a dermatological disorder with psychosocial implications on patients with hair loss. The objective of present study involves preparation of herbal hair oil using amla,brahmi,tridex,shikakai, neem,Antioxidant activity and its evaluation for increase in hair growth activity and other uses of antioxidants.Hair oils are the hair care preparations used for the prevention and treatment of baldness or other ailments, aggression of hair. They also promote luxurious growth hair. Hair oil containing herbal drugs are used as hair tonic. Hair care products are categorized in to main category.(1-2)

1-Hair Tonics.

2-Hair Grooming Aids.

These are basically the extracts of medicinal plants in an oil base. A plethora of herbs have been employed for hair treatment few of these are amla,  henna ,neem, methi ,lemon, tulsi, shikakai, reetha, liquorice root, musk root, mahabhringraj, jantamasi, chitraka, marigold, hibiscus, nutmeg and tridex.(8)

 Synthetic drug, minoxidil a potent vasodilator, appears safe for long term treatment. Oral minoxidil increase growth of body hair. Applied topically (2% twice daily) it promotes hair growth in male pattern baldness and alopecia areata. The response is slow (takes 2-6 months) and in complete, but up to 60% subjects derive some benefit, albeit for short periods. Baldness recurs when therapy is discontinued. The mechanism of increased hair growth is not known, may involve:

(a)Enhanced microcirculation around hair follicles.
(b)Direct stimulation of resting hair follicles.
(c)Alteration of androgen effect on genetically programmed hair follicles.

After five years use of 2 to 3% topical minoxidil, the improvement has been shown to peak at one year with a slow decline in regrowth over subsequent years. Long term treatment with local side effects may be a problem with continuing use of minoxidil lotion.(6)

On the basis of market survey carried out on crude drugs used presently for herbal hair oils gives us clue for selection of drugs for hair oil. Hence the present study was aimed to evaluate the hair growth activity of herbal formulations, which includes oil extract of all the mentioned drugs in various concentrations. In order to justify the traditional claims now a days multi ingredient hair oils are prepared and tested for their hair growth activity and antioxidant activity due tom this used other diseases.

Amla (Emblica officinalis) belong to family Euphorbiacae is rich in vitamin c , tannins and minerals such as phosphorus ,iron ad calcium which provide nutrition  to hair and also causes darkening of hair.(3)

Brahmi (bacopa monnierie) belong to family scrophulariaceae contains alkaloids which enhance protion kinase activity and also used for treatment for Dementia.(7)

NEEM (Azadirachta indica) belongs to family Meliaceae contains glycerides of saturated and unsaturated fatty acids. The main fatty acids are oleic and stearic acids. These oils are used in the treatment of dandruff in hair and relief in itching.(5)

Shikakaiits botanical name is Acacia Concinna.The bark contains saponin, which on hydrolysis yields lupeol,spinasterol and acacic acid lactone. The sugars identified are glucose, arabinose and rhamnose. It also contains hexacosanol.The saponin of bark shows spermicidal activity against human semen. The tender leaves, which are acidic, are used in chutneys. The leaves contain oxalic, tartaric, citric, succinic and ascorbic acids. They also contain two alkaloids calyctomine  and nicotine, besides rutin and an enzyme tartaric racimase. A new triterpenoid saponin having the basic skeleton of oleanolic acid.Other constituents present in the leaves are tannins, amino acids and proteins. its botanical name is Acacia Concinna,it is used as natural hair wash for sensitive scalps or to control dandruff.It promots hair growth,strengthen hair roots and lengthy beautiful hair(9)

Tridax procumbens Linn. (Compositeae), a weed found throughout India is employed as indigenous medicine for a variety of ailments including jaundice. It is commonly used in Indian traditional medicine as anticoagulant, antifungal and insect repellent; in bronchial catarrh,diarrhoea and dysentery. Moreover it possesses wound healing activity and promotes hair growth.Tridax procumbens is also dispensed as‘Bhringraj’, which has a great  reputation in Ayurvedic medicines for liver disorders.The hepatoprotective action has been demonstrated.We have reported recently the antioxidant property of Tridax procumbens ontheprotective potential of Tridax procumbens.(10)

2-Materials and methods:
The fruits of Embelica officinalis(amla),leaves of Bacopa monnieri(brahmi), fruits of Acacia concinna(shikakai) Were procured from local market of Allahabad Leaves of Azadirachta indica (neem) were collected from herbal garden of Christians school of pharmacy, faculty of health and medical sciences, indigenous, alternative system of medicine, SHIATS Allahabad.Fresh leaves of Tridex procumbens belong to family compositeae were collected in the month of September from the department of agriculture Allahabad SHIATS. The collected leaves were dried in the shade and then powder to a coarse consistency and stored in an air tight container at room temperature

3-Preparation:
The herbs used in the present study for making herbal hair oil were dried ,crushed and passed through sieve no.80 and olive oil was used as a base. The hair oil was prepeared using following method.

3.1-Boiling Direct method:

First is the direct boiling method in which the crudedrugs were powdered, weighed and directly boiled in olive oil with continuous stirring and heating until the drug had completely extracted in the oil base. The composition of the oil was 5 and 8% where 5gm and 8gm of the individual drugs;Embelica officinalis,Bacopa monnieri,Azadirachta indica,tridx procumbens,Acacia concinna.were mixed in 100ml of the olive oil.after extraction completed then they are filtered by muslin cloth individually.the herbal hair oil taken in beaker and covered by alluminium foil and keep in freezer in cool condition.(8)

4-Composition of poly hair oil formulation:

5-Evaluation of Herbal Oil Preparation:

The prepared oils were then subjected to physical.

5.1-Physical evaluation
In physical evaluation, parameters like specific gravity, pH, refractive index, viscosity and acid value were determined and the formulations were subjected to biological evaluation. The results of primary physical screening are given in below.

5.2-Evaluation of physical parameters:

Concentration → 5% and 8%

Specific gravity     0.9384,   0.9432                                 

pH      8.4, 7.5

Refractive index

1.472, 1.435

Acid value

2.18 1.558

6-ANTIOXIDANTS:
An antioxidant is a moleculecapable of inhibiting the oxidationof other molecules. Oxidation is a chemical reactionthat transfers electronsfrom a substance to an oxidizing agent. Oxidation reactions can produce free radicals. In turn, these radicals can start chain reactionsthat damage cells. Antioxidants terminate these chain reactions by removing free radical intermediates, and inhibit other oxidation reactions. They do this by being oxidized themselves, so antioxidants are often reducing agentssuch as thiols, ascorbic acidor polyphenols.

 A low total antioxidant capacity (TAC)1 is considered a risk factor for the development of diseases such as cancer and cardiovascular disease . For antioxidants to be protective against disease, there must be adequate levels to prevent free radical damage. Oxidative stress occurs when the

free radical levels exceed the antioxidant stores.Antioxidants are added to a variety of foods to prevent or deter free radical-induced lipid oxidation, which is responsible for the development of off-flavors and the undesirable chemical compounds in food (Angelo, 1996). The free radicals can also be generated in biological systems in the form of reactive oxygen species (ROS), such as superoxide anion radicals (O2•−), hydrogen peroxide (H2O2), hydroxyl radicals (OH•), and the singlet oxygen (1O2) (Halliwell et al.,1995). These reactive ROS cause destructive and irreversible damage to the components of a cell, such as lipids, proteins and DNA (Lopaczyski and Zeisel, 2001). Although normal cells possess antioxidant defense systems against ROS, the continuous accumulation of damage to the cells induces diseases such as cancer and aging (Matés and Sánchez-Jiménez,2000). The continuous antioxidant dose also plays a preventive role against these diseases by removing the ROS in biological systems (Sgambato et al., 2001).Anticancer agents, on the other hand, are mainly related to their curative role in a damaged system. Under normal conditions,the cells in which the DNA or other components are irreversibly damaged by various causes undergo apoptotic cell death, which is a self destructive metabolism according to the genetically encoded cell death-signal (Korsmeyer,1995; Hooper et al., 1999).However, cancer cells, which are already irreversibly developed, obtain the capability to evade apoptosis by various ways.The aim of anticancer agents is to trigger the apoptosis signaling system in these cancer cells whilst disturbing their proliferation (Bold et al., 1997).

7-DPPH radical scavenging test:
This test was measured as described by Blois (1958). Onemillilitre of the extract (oil) solutions (10, 20, 30, 40 and 50 micro gm/ml.in acetone) was added to 1mL of a DPPH solution (0.2mMin acetone). All the fractions showed dose-dependent increase in activity. After a 30 min of reaction at room temperature, the absorbance of the solution was measured at 517 nm. The free radical scavenging activity of each fraction was determined by comparing its absorbance with that of a blank solution (no sample). The Antioxidant activity the oil is measured against ascorbic acid (94% Antioxidant activity).

7.1-Scavenging activity of free radical:
Free radicals are known to be a major factor in biological damages, and DPPH has been used to evaluate the free radical-scavenging activity of natural antioxidants (Yokozawa et al., 1998; Zhu et al., 2001). DPPH, which is a radical itself with a purple color, changes into a stable compound with a yellow color by reacting with an antioxidant,and the extent of the reaction depends on the hydrogen donating ability of the antioxidant.

Antioxidant activity (AA) was expressed as percent of inhibition relative to the control, using the following formula:

                                       AA = ×100

 2,2 -diphenyl-2-picrylhydrazyl radical scavenging assay.

The scavenging effect on of the DPPH radical was calculated using the following equation:

                  Scavenging effect % =    × 100

EC50 value was determined from the plotted graph of scavenging activity against the concentration of cocoa extracts, which is defined as the total antioxidant necessary to decrease the initial DPPH radical concentration by 50%.Triplicate measurements were carried out, and their scavenging effect was calculated based on the percentage of DPPH scavenged.

Control limit for DPPH = 0.823

8-Drug extract absorbance:

 9-Result:
The prepared formulation of hair oil is showing good antioxidant activity when compared with standard (ascorbic acid). Our results revealed that the antioxidant capacity of the herbal hair oil is comparable to the antioxidant activity of Ascorbic acid. Although there are various methods are available for measuring antioxidant activity, we used DPPH method as standard one.

10-Conclusion:
Our study showed that the herbal hair oil is showing good antioxidant activity. TheAntioxidant property of plant and oil can be  utilise in hair fall as well as in the treatment of cancer and various other diseases.Antioxidants are widely used as ingredients in dietary supplementsand have been investigated for the prevention of diseases such as cancer, coronary heart diseaseand even altitude sickness.our herbal oil show good antioxidant activity  so it can be used various diseases Like Free radical damage within cells has been linked to a range of disorders including cancer, arthritis, atherosclerosis, Alzheimer's disease, and diabetesand many other diseases.

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Reference ID: PHARMATUTOR-ART-1013

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