Indian Journal of Experimental Biology Vol. 50, November 2012, pp. 785-794 Nanoemulsified ethanolic extract of Pyllanthus amarus Schum & Thonn ameliorates CCl 4 induced hepatotoxicity in Wistar rats V Deepa 1 *, R Sridhar 1 , A Goparaju 2 , P Neelakanta Reddy 3 & P Balakrishna Murthy 4 1 Department of Biotechnology, 2 Department of Statistics & 4 Department of Toxicology, International Institute of Biotechnology and Toxicology, Kanchipuram District, Padappai 601 301, India, 3 Bio-organic Chemistry Laboratory, CSIR-Central Leather Research Institute, Adayar, Chennai 600 020, India Received 4 November 2011; revised 27 August 2012 Phyllanthus amarus (PA) is commonly used in traditional medicine for hepatoprotectivity. The major limitation is that, treatment requires a large quantity of herbal extract for a longer duration. Aim of the present study was to encapsulate ethanolic plant extract for sustained release of constituents in intestine and facilitate maximum absorption. The efficacy was compared for the hepatoprotective activity of nanoencapsulated ethanolic extract of P. amarus (NPA) and PA in carbon tetrachloride (CCl 4 ) induced hepatotoxic male rats. Based on total phenol content (TPC), the loading efficiency of nanocapsules was 89% (pH 7.0) and optimum concentration was 2:18 (mg/mL) for plant extract: olive oil. Scanning electron microscopy (SEM) showed a spherical morphology, photon correlation spectroscopy (PCS) identified mean particle diameter as 213 nm and Fourier transform infrared spectroscopy (FT-IR) revealed that the phytoconstituents were stable. An oral dose of NPA (20 mg/kg body wt.) showed a better hepatoprotective activity than PA (100 mg/kg body wt.) and also repeated dose oral toxicity proved to be safe. These biochemical assessments were supported by rat biopsy examinations. In conclusion, the nanoemulsification method may be applied for poor water-soluble ethanolic herbal extracts to reduce the dosage and time. Keywords: Hepatoprotective activity, Nanoemulsion, Phyllanthus amarus, Sodium alginate Phyllanthus amarus is a herb used in Ayurvedic medicine for liver disorders 1 . It is a well-known antiviral agent 2 , has antiseptic properties, and is also used to treat gonorrhea, jaundice and mammary abscesses 3 . In addition, P. amarus has been reported to possess antidiabetic, anticancer and anti-inflammatory activities 4-6 . It contains many types of phenolic compounds; lignans such as phyllanthin and hypophyllanthin, flavonoids such as quercetin and astragalin and ellagitannins such as amarinic acid, amarin and phyllanthin D 7-10 . These phenolic compounds were related to P. amarus antioxidant activity 11 . Biochemical constituents of plants are important sources of natural antioxidants and efficacy of plant extract is more when they are consumed as a crude extract 12 . However, a major limitation is that the quantity of herbal extract required for treatment is more and a long duration of treatment is required due to the degradation of various plant constituents such as alkaloids, amides, prophenylphenols, steroids, hydrocinnamic acid and oxalic acid in gastro intestinal tract, in addition to poor absorption of these constituents in intestine 13 . In this study, nanocapsules of P. amarus were prepared by emulsion-coacervation method and characterized them by using scanning electron microscopy (SEM), photon correlation spectroscopy (PCS) and Fourier transform infra-red spectroscopy (FT-IR). Finally, the efficacy of nanoemulsified ethanolic extract of P. amarus was studied for their hepatoprotective activity by reducing the treatment dosage and period. Materials and Methods Extraction and preparation of nanoemulsified ethanolic extracts using sodium alginate—Leaves and berries of P. amarus were collected from International Institute of Biotechnology and Toxicology, Padappai, during the period 2009. They were washed thoroughly with several changes of sterile water followed by 70% ethanol, blotted between fields of filter paper ——————— *Correspondent author Telephone: + 91-44-27174246, + 91-44-27174266 (Inst) Fax: + 91-44-27174455 E-mail: deepasabarish@gmail.com