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Research Article In vitro antioxidant and standardization studies on formulated hair oil AG2 Sudha Veerappan, Niraimathi Lakshmanan*, Lavanya Ramu, Brindha Pemaiah ABSTRACT Objective: The objective of the study was to develop herbal hair oil using leaves of Alternanthera sessilis L. and root powder of Glycyrrhiza glabra. Method: Preliminary phytochemical screening was performed for the herbal hair oil in the detection of secondary metabolites. The formulated oil was assessed by general physicochemical properties including color, odor, density, acid value, saponification value, peroxide value, and rancidity. In addition, gas chromatography-mass spectrometry (GC-MS) analysis was performed to identify the active metabolites in the formulation. In vitro antioxidant activity of prepared herbal hair oil was determined by (2, 2- Diphenyl-1-(2, 4, 6-trinitrophenyl) hydrazyl) radical scavenging assay. Results: GC-MS analysis detected 24 compounds from herbal formulation which exhibit various types of fatty acids, terpenoids and hydrocarbons. The major fatty acids found in the formulation were lauric acid (36.09%), myristic acid (19.97%), oleic acid (12.49%) and palmitic acid (11.81%). Antioxidant activity of formulated oil was determined and its IC50 value was found to be 50.84% at the concentration of 400µg/ml using gallic acid as standard. Conclusion: The result suggested that the selected herbs A. sessilis and G. glabra in coconut oil can contribute significantly in promoting hair growth. Chemical moieties identified might be contributing to the antioxidant efficacy of the developed herbal hair oil AG2. KEY WORDS: Antioxidant activity, gas chromatography-mass spectrometry, hair oil, standardization INTRODUCTION Coconut oil is the most popular oil used for hair care around the world. The chemical compounds present Herbal cosmetics are in high demand due to the in the coconut oil such as saturated fatty acids are increasing interest of mankind toward them because responsible for hair growth.[2] Saturated fatty acid is they are more effective with less side effects and one of the richest sources of medium chain fatty acids. easily available ingredients. Hair care cosmetics are The major fatty acids are lauric acid, myristic acid, now used with herbs as they are well recognized and and capric acid.[3] The fatty acids help the coconut oil human-friendly as compared to synthetic ones. Hair to penetrate the hair shafts easily and supply essential oil preparations mainly used for the prevention and nutrients required by the hair and also help to make treatment of baldness or other ailments, the aggression the hair more radiant, thick, and healthy. Lauric acid, of hair. They also promote the luxurious growth of hair. which is highly abundant in coconut oil can eradicate Over centuries, the popular method to stimulate hair the microbes such as bacteria, fungi, and virus.[4] Hair growth seems to be to use herbs and oils. The Egyptians is totally made up of proteins, and it gets reduced applied castor oil directly to the scalp with the hope of due to overuse of chemical based hair products and stimulating hair growth. To enhance the smell, they leads to weak, broken, and unhealthy hair. Coconut sometimes mixed it with sweet almond oil. Ancient oil is the only hair growth oil that serves as a protein Indians and Polynesians applied coconut oil directly to shield for the hair, and the fatty acids have the power the scalp whereas Ancient Africans used olive oil. Based to reduce this protein loss from the hair. Coconut oil on a previous study, it appears that certain plant-based helps to prevent the loss of moisture from hair and thus [1] oils have a positive impact on hair loss treatment. provides moisturizing effect. Coconut oil is also a great source of antioxidants, hair vitamins, and nutrients. Access this article online These hair vitamins prevent hair fall, escalate faster hair growth and strengthens, softens, and contributes Website: jprsolutions.info ISSN: 0974-6943 more effectively than any other hair growth oil. Centre for Advanced Research in Indian System of Medicine (CARISM), SASTRA Deemed to be University, Thanjavur, Tamil Nadu, India *Corresponding author: Niraimathi Lakshmanan, Centre for Advanced Research in Indian System of Medicine, SASTRA University, Thanjavur – 613 401, Tamil Nadu, India. Phone: +91-9944690268. E-mail: [email protected] Received on: 17-12-2017; Revised on: 25-01-2018; Accepted on: 14-03-2018 Journal of Pharmacy Research | Vol 12 • Issue 3 • 2018 431 Sudha Veerappan, et al. An excessive amount of herbs have been employed for In the present study, herbal hair oil was prepared using hair treatments. A few of these herbs are Phyllanthus coconut oil as a base and leaf juice of A. sessilis L and emblica,[5] Lawsonia inermis, Azardirachta indica, G. glabra root powder. The developed formulation Trigonella foenum graecum, Citrus limon, Ocimum was standardized and evaluated for its in vitro sanctum, Bacopa monnieri, Acacia concinna, antioxidant activity. Sapindus mukorossi, G. glabra root, Eclipta prostrata, Nardostachys jatamansi, Hibiscus rosa sinensis, and METHODS Thymus vulgaris.[6,7] In the present study, two herbs such as sessile joywood and G. glabra root are used Plant Material along with coconut oil. A. sessilis leaves were purchased from local market, Thanjavur and G. glabra root from Siddha Sessile joywood is botanically equated as Alternanthera pharmacy store, Thanjavur and were authenticated sessilis (L.) R.Br. ex DC. is an aquatic plant belongs by Dr. N. Ravichandran, CARISM, SASTRA deemed to family Amaranthaceae. It is known by several University, Thanjavur, Tamil Nadu. common names including ponnanganni (in Tamil). As an herbal medicine, the plant has diuretic, cooling Chemicals and Reagents hair tonic, and laxative properties. It has been used for 2, 2- Diphenyl-1-(2, 4, 6-trinitrophenyl) hydrazyl the treatment of dysuria and hemorrhoids.[8] This herb (DPPH) was purchased from Sigma-Aldrich, USA. has been reported to be antioxidant, antimicrobial, All other chemicals were of analytical reagent grade and wound healing.[9] The plant is also believed to be purchased from Merck Specialties Pvt., Ltd., India. beneficial for the eyes and used as an ingredient in the preparation of medicinal hair oils. Application of Preparation of Herbal Hair Oil coconut oil boiled with A. sessilis leaves, Hibiscus, and The hair oil was prepared using A. sessilis leaves, Cynodon dactylon helps in luxurious hair growth. The G. glabra root powder and coconut oil. A. sessilis leaves of A. sessilis are rich in alkaloids, flavonoids, leaves were grind well with water. The obtained leaf terpenoids, carbohydrates, saponins, tannins, steroids, juice was filtered. Equal volume of filtered juice and starch, protein, amino acids, Vitamin C, and contains coconut oil was boiled with G. glabra root powder high amount of calcium and iron.[10] Flavonoids and with constant stirring at a constant temperature until triterpenoids possess hair growth promoting activity the water droplet in oil stopped with crackling sound by strengthening the capillary wall of the smaller and the drugs were completely extracted in the oil. blood vessels present in hair follicles and improves After complete extraction, the oil was filtered using blood circulation so as to nourish the hair follicles and muslin cloth and stored. The formulated herbal hair oil thereby promoting hair growth.[11,12] was subjected to chemical standardization studies and antioxidant activity. G. glabra belonging to the family Fabaceae is a plant which grows in Egypt and other countries of Standardization of Herbal Hair Oil the world. Its roots possess some nutritive value and The formulated oil was screened chemically for the medicinal properties. G. glabra plant is an herbaceous identification of chemical constituents and the quality perennial legume native to southern Europe and parts of the oil was evaluated by standard methods.[15] of Asia, such as India. Their roots are thick having many branches with red or lemon color outside and Chromatographic Analysis yellowish or pale inside. It is called Keshya (hair Gas chromatography-mass spectrometry (GC-MS) growth promoter) in traditional texts of Ayurveda.[13] analysis was performed using a gas chromatograph G. glabra root is said to be very soothing for dry, equipped with Turbomass Quadrupole Mass irritated skin. Many of the properties found in the Spectrometer (PerkinElmer, Clarus 500, USA) fitted root are capable of soothening the scalp and getting with Elite-5 capillary non-polar column (30 m × 0.25 mm rid of irritations such as dandruff and scabs. It is internal diameter, film thickness 0.25 µm). An electron mollifying properties can help in open up the pores ionization system with ionization energy of 70ev was of the scalp and are beneficial for the hair shaft. used for the detection of chemical constituents. Helium Chemical analysis of G. glabra root extract showed gas with 99.999% purity was used as a carrier gas at the existence of triterpenes, flavonoids, and a wide constant flow rate of 1 ml/min. The injector temperature range of beneficial nutrients.[14] It is a good source of was set at 280°C and the oven temperature was set Vitamin B1 (thiamine), B2 (riboflavin), B3 (niacin), initially at 60°C–150°C at 6°C/min hold for 2 min and B5 (pantothenic acid), and Vitamin E (tocopherol). It finally raised to 280°C at 4°C/min hold for 5 min. The also provides minerals such as phosphorous, calcium, sample was diluted in n-hexane and 1 µl was injected choline, iron, magnesium, potassium, selenium, with split ratio of 1:10. The mass spectrum was scanned silicon, and zinc. from 40 to 450 amu with scan time interval of 0.2 s. The 432 Journal of Pharmacy Research | Vol 12 • Issue 3 • 2018 Sudha Veerappan, et al. percentage of the chemical constituents in the sample inhibition of sample at different concentrations was expressed as % area by peak area normalization.