GC-MS and Phytopharmacological Analysis of Aqueous Distillate of Boerhavia diffusa Roots

Objective: To investigate the phytochemical constituents of aqueous distillate of Boerhavia diffusa roots by using GC-MS technique.Methods: GC-MS analysis of aqueous distillate of Boerhavia diffusa roots were performed using a Perkin-Elmer GC Clarus 500 system and Gas Chromatograph interfaced to a Mass Spectrometer (GC-MS) equipped with a Elite -I, fused Silica Capillary Column (30mm x0.25mm1Dx1 μ Mdf, composed of 100 % Dimethylpolysiloxane. Results: In this study results showed the presence of many phytochemical compounds such as phenol, lipid, saponin, flavonoids, steroids, protein, and carbohydrates. In GC-MS analysis, 31 bioactive Phytochemical compounds were identified in the aqueous distillate of Boerhavia diffusa roots, the components were identified by comparing their relation indices and mass spectra fragmentation patterns with those stored on the MS-Computer library and also from the published literatures. Stigmast-4-en-3one 48.066% followed by Cycloartane-3.beta., 25-diol (RT) 46.226%, gamma-Sitosterol with RT 44.193, Stigmasterol with 42.788% RT, Dihydrobrassicasterol with RT 42.074%, Lichesterol RT 41.647%, Cholesteryl chloroformate RT 39.518 % were seem to be the major constituents. Conclusion: From the results, it can be concluded that the aqueous distillate of Boerhavia diffusa roots showed 31 bioactive phytochemical constituents. The presence of these constituents justifies the use of plant roots by folk practitioners. Since terpenoids and sterols are the basic components responsible for antioxidant, antibacterial, anti-inflammatory activity and ailments of many diseases, we identify the presence of terpenoids and sterols in aqueous distillate of Boerhavia diffusa roots as major component, hence it may be used in various diseases. Pooja*, V. K. Lal, Anurag Verma School of Pharmaceutical Sciences, IFTM University, Moradabad, U.P., India. Submission: 25 August 2017 Accepted: 3 September 2017 Published: 30 September 2017 www.ijppr.humanjournals.com Citation: Pooja et al. Ijppr.Human, 2017; Vol. 10 (2): 374-391. 375 INTRODUCTION: As mentioned in ancient Indian books like Charak Samhita, Sushruta Samhita, Ras Tantra Sar, Bhavprakasha, Ras Tarang, Nayan Drastam and Astanghriday, Sushruta, the father of Surgery in India before 500 BC researched that, there are a number of plants which are used in various diseases, As juice, extract, kwath, paka, arka etc, either single or in compound formulations with other plants 1 . Medicinal plants have no side effects due to presence of bioactive constituents. Medicinal plants are of great importance to the health of individuals and communities in general 2 . The medicinal value of plants lies in some chemical substances that produce a definite physiological action on the human body. The most important of these bioactive constituents of plants are alkaloids, tannins, flavonoids and phenolic compounds 3 . In Ayurveda (Indian system of medicine) various disorders and diseases have been described in great details. The World Health Organization (WHO) supports the use of traditional medicine, they are proven to be efficacious and safe 4 . Many of the indigenous medicinal plants are used as spices and food. They also sometimes added to foods meant for pregnant women and nursing mothers for medicinal purposes 1-3 . In addition, the use of herbal medicine for the treatment of diseases and infections is as old as mankind 4 . Boerhavia diffusa is a member of Nyctaginaceae family. Boerhavia diffusa is a perennial creeping weed; a number of branches spread out from the node of main stem and cover most of the surrounding area. The stem is usually prostrate and woody. Leaves are simple, ovate or oblong or round in shape flowers are minute, hermaphrodite, subcapitates, pedicellate. Stamens are usually two or three in number and the stigma is peltate. The fruit is achene, ovate, oblong, and pubescent. Five ribbed and glandular and anthocarpous 5, 6, 7 . Taxonomical classification, common names and photographs of leaves, flower and plant of (Boerhavia diffusa) is given in figure 1. www.ijppr.humanjournals.com Citation: Pooja et al. Ijppr.Human, 2017; Vol. 10 (2): 374-391. 376 Fig. 1: Taxonomical classification, common names and photographs of leaves, flower and plant of Boerhavia diffusa Part used: Root, Leaves and seeds 7 GEOGRAPHICAL DISTRIBUTION AND HABITAT: Genus Boerhavia, consisting of 40 species is distributed in tropical and subtropical regions and warm climate. It is found in Ceylon, Australia, Sudan and Malay Peninsula, extending to China, Africa, America and Islands of the Pacific. Among 40 species of Boerhaavia, 6 species are found in India, namely B. diffusa, B. erecta, B. rependa, B. chinensis, B. hirsute and B. rubicunda. Boerhavia diffusa, in India, is found in warmer parts of the country and throughout up to 2000 m altitudes in the Himalayan region. It is a perennial, spreading hogweed, commonly occurring abundantly in waste places, ditches and marshy places during rains. The plant is also cultivated to some extent in West Bengal 8, 9, 10, 11 . Boerhavia diffusa is a perennial creeping weed, prostrate or ascending herb, up to 1 m long or more having spreading branches. The plant grows profusely in the rainy season, and mature seeds are formed in October-November. Due to its sticky nature, the plant gets stuck on the clothes of human beings and on the legs of animals, which helps in its dispersal from one place to another 12 . The shape of the leaves varies considerably ovate-oblong, round, or subcordate at the base and smooth above. Margins of the leaves are smooth, wavy, or undulate. The upper surface of www.ijppr.humanjournals.com Citation: Pooja et al. Ijppr.Human, 2017; Vol. 10 (2): 374-391. 377 the leaves is green, smooth, and glabrous, whereas it is pinkish white and hairy beneath. Leaves are up to 5.5 × 3.3 cm2 in area. The seeds germinate before the onset of the monsoon 13 . Flowers are minute, usually fascicled or sub umbellate on the ultimate branchlets, pink, white and about 1.5 mm long. These are hermaphrodite, pedicellate, and white, pink, or pinkish-red in color. Bracts are deciduous and involucrate. A perianth is present in the place of a calyx and corolla, which is tubular in shape, the tube being short and narrow at the base and funnelshaped at the top and constricted above the ovary. There are five lobes, which are small and acute. Two or three stamens are present and are slightly exerted. The stigma is peltate. The achene fruit is detachable, ovate, oblong, pubescent, five-ribbed and glandular, anthocarpous, and viscid on the ribs 6, 7, 8, 13. The taproot is tuberous, cylindrical to narrowly fusiform to conical or tapering, light yellow, brown or brownish gray. It is thick, fleshy and very bitter in taste. Some workers have studied the regeneration of this plant through tissue culture 14 . Roots contained 0.15% alkaloid punarnavine. Increase in levels of indoleacetic acid (IAA) in MS medium reduced the number of roots regenerated from the leaf segment, their length and alkaloid content 15 . The present study clearly demonstrated that the aqueous distillate of Boerhavia diffusa roots still possesses some useful phyto compounds analyzed by GCMS, which have many pharmacological properties and may provide maximum information for the use in medicine 16 . MATERIALS AND METHODS: Collection of plant materials: Well identified samples of plant Boerhavia diffusa Linn. (roots) were collected from their natural habitats along with the samples from local market 17 . The samples have been authenticated by National Botanical Research Institute, Lucknow. (SPECIFICATION No: NBRI-SOP-202). The plants were washed in tap water and air dried. Roots were collected in separate paper covers and dried in shade for 15 days 18 . The dried plant materials (roots) of (Boerhavia diffusa) made to fine powder using homogenizer. The dry powder obtained was stored at room temperature in airtight containers 19 . www.ijppr.humanjournals.com Citation: Pooja et al. Ijppr.Human, 2017; Vol. 10 (2): 374-391. 378 METHOD OF PREPARATION: The drug was cleaned and coarsely powdered. Some quantity of water is added to the drugs for soaking and kept over-night. This makes the drugs soft and when boiled releases the essential volatile principles easily. Powdered root of plant was boiled with distilled water (1:16) where 1 part is drug and 16 parts are distilled water. The following morning it is poured into the distillation assembly and the remaining water was added and boiled. The vapour was condensed and collected in a receiver. In the beginning, the vapour consists of only steam and may not contain the essential principles of the drugs. It should, therefore, be discarded. The last portion also may not contain therapeutically essential substance and was discarded. The aliquots collected in between contain aroma of the active ingredients and may be mixed together to ensure uniformity of the aqueous distillate 8 . Condensed aqueous distillate of roots, were stored in airtight container, and used for GCMS study and preliminary screening of phytochemicals such as alkaloids (Mayer's and Dragendorff’s tests), anthroquinones (Borntrager's test), flavonoids (NaOH or Alkaline reagent test), cardiac glycosides (Keller-Killani test), phenols (Phenol test), quinines (NaOH test), reducing sugars (Fehling's test), saponins (Foam test), steroids (Libermann-Burchard test), soluble starch (Iodine test), tannins (gelatin test) and terpenoids (Salkowski test), were carried out following the methodologies of Harbone (1998), Sofowora (1993), Trease and Evans (2002). Standardization of aqueous distillate 1. Preliminary Phytochemical screening of aqueous distillate for the presence of group of compounds The aqueous distillate so prepared has been tested qualitatively for the presence of different groups of compounds and details are as below 10 . www.ijppr.humanjournals.com Citation: Pooja et al. Ijppr.Human, 201