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Comparative Phytochemical Analysis and Antibacterial Efficacy of in Vitro and in Vivo Extracts from East Indian Sandalwood Tree

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Comparative Phytochemical Analysis and Antibacterial Efficacy of in Vitro and in Vivo Extracts from East Indian Sandalwood Tree See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/231610036 Comparative Phytochemical Analysis and Antibacterial Efficacy Of In Vitro And In Vivo Extracts From East Indian Sandalwood Tree. ARTICLE in LETTERS IN APPLIED MICROBIOLOGY · SEPTEMBER 2012 Impact Factor: 1.66 · DOI: 10.1111/lam.12005 · Source: PubMed CITATIONS READS 11 205 2 AUTHORS: Biswapriya Biswavas Misra Satyahari Dey University of Florida IIT Kharagpur 52 PUBLICATIONS 110 CITATIONS 69 PUBLICATIONS 667 CITATIONS SEE PROFILE SEE PROFILE All in-text references underlined in blue are linked to publications on ResearchGate, Available from: Biswapriya Biswavas Misra letting you access and read them immediately. Retrieved on: 21 March 2016 Letters in Applied Microbiology ISSN 0266-8254 ORIGINAL ARTICLE Comparative phytochemical analysis and antibacterial efficacy of in vitro and in vivo extracts from East Indian sandalwood tree (Santalum album L.) B.B. Misra1,2 and S. Dey1 1 Plant Biotechnology Laboratory, Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur, Midnapore (West), West Bengal India 2 Present address: Center for Chemical Biology, Universiti Sains Malaysia [CCB@USM], 1st Floor Block B, No 10, Persiaran Bukit Jambul, 11900 Bayan Lepas, Pulau Pinang, Malaysia Significance and Impact of Study: This study constitutes the first systematic investigation on phytochem- ical composition and antimicrobial efficacy of sandalwood tree across in vitro and in vivo developmen- tal stages screened against thirteen bacterial strains by four methods. Using a battery of antimicrobial assay techniques, it is possible to follow the differential bioactive metabolic richness of plant parts, to decipher, for example comparable efficacy of somatic embryo extracts and sandalwood oil. Keywords Abstract antibacterial, extracts, in vitro, phytochemical, sandalwood oil, santalol, Santalum album, Sandalwood oil has been found in numerous therapeutic applications in TLC bioautography. traditional medicines such as Chinese traditional medicine and Ayurveda. However, there are no comparative accounts available in the literature that Correspondence focused on in vitro and in vivo tree sample-derived extracts. Combined B.B. Misra, Plant Biotechnology Laboratory, dichloromethane and methanol extracts were obtained from in vitro samples, Department of Biotechnology, Indian Institute that is, callus, somatic embryo and seedlings, and in vivo from leaves of non-oil- of Technology Kharagpur, Midnapore (West) Kharagpur-721302, West Bengal, India. yielding young and oil-yielding matured trees. Phytochemical evaluation of the E-mail: [email protected] extracts reveals that the tree is rich in terpenoids, saponin, phenolics and tannins. The antibacterial properties of the five extracts were compared with sandalwood 2012/1333: received 25 July 2012, revised 6 oil by screening against nine Gram-negative and five Gram-positive bacterial September 2012 and accepted 23 September strains by disc diffusion, agar spot and TLC bioautography methods. Minimum 2012 inhibitory concentration (MIC) for sandalwood oil was determined to be in the À range of 0Á078–5 lgml 1 for most of the test micro-organisms screened. doi:10.1111/lam.12005 Bioautography results indicated the presence of potential antimicrobial constituents in somatic embryo extracts and sandalwood oil. Among the extracts screened, the somatic embryo extracts showed the strongest antibacterial activity comparable only with sandalwood oil and matured tree leaves’ extract. The findings presented here also suggest that apart from sandalwood oil, other parts of this tree across developmental stages are also enriched with antibacterial principles. matured tree ranges from 2Á5 to 6%, which is the highest Introduction among all known 20 Santalum species, depending on the The East Indian sandalwood tree, Santalum album L., is age of the tree and colour of the heartwood (Shankaran- identified as one of the most important medicinal tree in arayana and Kamala 1989). The estimated global annual the world (Fox 2000). This woody tree is a root hemipara- requirement is about 10 000 tons of wood, equivalent to site tropical species belonging to the taxonomic group 200 tons of oil, involving a trade of about $360 million, of Santalaceae and yields precious sandalwood, which con- which only 10% is met from natural resources. The quality tains over 90% of santalols, a mixture of sesquiterpenoids and composition of essential oil obtained from young and and hence is the focus of many investigations (Demole matured sandal trees varies among heartwoods and sam- et al. 1976). The essential oil yield from a 50-year-old pling heights (Shankaranarayana and Parthasarathi 1987). 476 Letters in Applied Microbiology 55, 476--486 © 2012 The Society for Applied Microbiology B.B. Misra and S. Dey Antibacterial principles of sandalwood Sandalwood oil is used in Indian traditional medicine extracts (8–10%, w/w) and was the lowest in callus at 4%. system Ayurveda, as antiseptic, antipyretic, antiscabietic, The ash content of the five tissues showed variability diuretic, expectorant, stimulant and for the treatment of depending upon their in vitro and in vivo origin. bronchitis, dysuria, urinary infection, gonorrhoeal recov- ery owing to its antibacterial and antifungal properties Phytochemical evaluation of sandalwood extracts (Handa et al. 1951; Okasaki and Oshima 1953; Winter 1958; Dastur 1962; Jain 1968; Dikshit and Hussain Phytochemical analyses suggest that the extracts were rich 1984). The hydrolysed exhausted sandalwood powder on in terpenoids (monoterpenoids and sesquiterpenoids), pharmacological screening demonstrated antiremorogen- saponins, phenylpropanoids such as phenolics, proantho- ic, anti-inflammatory, antimitotic, antiviral, anticancer- cyanidins, flavonoids, phenolics, condensed tannins, o- ous, anti-hypertensive, antipyretic and sedative properties quinines and polyphenols (Table S1). (Desai et al. 1991). The oil also possesses antiviral activ- Sesquiterpenoid standard curve (y = 0Á002x, ity against certain herpes simplex virus types (Benencia R2 = 0Á988) was obtained for quantification purpose and Courreges 1999). Recently, the major constituent of using bulk-purified a-santalol as a standard. Total sesquit- oil, a-santalol, was proven to be effective as a skin can- erpenoid content in in vitro plant extracts (17Á3– À cer chemopreventive agent (Kaur et al. 2005). Moreover, 51Á4mgg 1 extract) was found to be higher than that of À it was shown that various organic fractions and sesquit- in vivo extracts (8–37Á4mgg 1 extract). Total monoterp- erpenoid compounds from the oil possess anti-Helicob- enoid content was quantified using an l-linalool acter pylori properties, which is the causative organism (y = 0Á0001x, R2 = 0Á932) standard curve and was found À for gastric cancer and peptic ulcer (Takaishi et al. 2005). to be similar for the extracts (3–4Á5mgg 1 extract) and À Epidemic phytoplasmal ‘spike’ disease leading to severe the highest for young tree leaves’ extract (9Á5mgg 1 destruction, illegal poaching and over exploitation owing extract). Total saponin content (diosgenin standard curve, to the increased global demand are the reasons of it being y = 0Á017x À 0Á197, R2 = 0Á988) of callus, somatic inducted into IUCN, Red List of Threatened Species as embryo and seedling extracts was comparable (9Á4– À vulnerable (IUCN 2006). Unsurprisingly, the first in vitro 17Á1mgg 1 extract), whereas the in vivo extracts showed À micropropagation study on any woody forest plant was higher saponin contents (31Á6–43Á6mgg 1 extract). reported for sandalwood, that is, callusing from embryos Phenolics are the most plentiful classes of constituents (Rangaswamy and Rao 1963) followed by somatic in the plant kingdom (Rao 2003). In sandalwood phy- À embryogenesis, regeneration, suspension cultures, somatic toextracts, 18–190 mg gallic acid equivalents (GAE) g 1 embryo production and maturation in air lift bioreactors of total phenolics was recorded. The highest phenolics (Das et al. 1998). Furthermore, in vitro callus is known content recorded for dietary constituents is spinach À to yield sandalwood oil constituents (Yamashita 1997). (c.0Á71 mg g 1), followed by swamp cabbage À À With the developmental stages of the plant very well (c. 0Á41 mg g 1), and cabbage (c. 0Á11 mg g 1; Ismail established and its age old medicinal properties being et al. 2004). Tannins are widely distributed among the confirmed recently by advanced means, this study was angiosperms. Furthermore, tannins such as gallic acid, undertaken to probe the antibacterial properties of non- catechins and tannin-glycosides were reported in the cal- polar extracts from in vitro samples, that is, callus, lus cultures of Quercus acutissima (Tanaka et al. 1995). somatic embryo and seedlings, and in vivo from the The fact that saponins are represented in sandalwood À leaves of non-oil-yielding and oil-yielding matured tree extracts (9Á4–43Á6mgg 1 extract) is further corroborated leaves with sandalwood oil. by the fact that the plant order Santalales are known to To our knowledge, this is the first time effort towards contain oleane-type triterpenoid saponins (Crespin et al. the evaluation of biological activities of the nonpolar 1993). In grape marc extracts, anthocyanins are present in À extracts from this particular tree, as a comparative study 0Á68 mg g 1 extract (Vatai et al. 2008), comparable with À across both in vitro and in vivo stages. the anthocyanin
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