Screening and quantification of an antiseptic alkylamide, spilanthol from in vitro cell and tissue cultures of Spilanthes acmella Murr.

Department of Biotechnology, Indian Institute of Technology - Guwahati, Guwahati 781039, Assam, India
Industrial Crops and Products (Impact Factor: 2.84). 03/2012; 36(1):321–328. DOI: 10.1016/j.indcrop.2011.10.029


The study revealed, for the first time, accumulation of spilanthol, an antiseptic alkylamide, in in vitro cultures of Spilanthes acmella Murr., a medicinal plant of immense commercial value. To achieve this, in vitro shoots were regenerated via direct organogenesis from leaf-disc explants of Spilanthes. Shoots were induced in the presence of N6-benzylaminopurine (BAP) alone or in combination with either α-naphthalene acetic acid (NAA) or Indole-3-acetic acid (IAA) in Murashige and Skoog medium. The best treatment for shoot regeneration was MS + BAP (5.0 μM) + IAA (5.0 μM), which promoted adventitious shoot proliferation in >82% cultures with an average of 5.3 shoots per explant. Regenerated shoots rooted spontaneously with a frequency of 100% on half strength MS medium (major salts reduced to half strength) containing 50 g l−1 sucrose. The plantlets were acclimatized successfully with 90% survival rate. Additionally, ploidy stability of the regenerated plants was assessed by flow cytometry which showed that all investigated plants had the similar ploidy as that of the mother plant. For spilanthol identification, peaks eluted from HPLC were analyzed by mass spectrometry with its characteristic fragmentation pattern. For quantification studies, calibration curve was generated, which revealed a higher amount of spilanthol content (3294.36 ± 12.4 μg/g DW) in the leaves of in vitro plants compare to those of in vivo plants (2703.66 ± 9.6 μg/g DW of spilanthol). An efficient multiplication frequency, ploidy stability and enhanced spilanthol accumulation ensure the efficacy of the protocol developed for this industrially important medicinal plant.

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Available from: Rakhi Chaturvedi,
    • "03 ± 15 . 6 mg / g DW ) ( Singh and Chaturvedi 2012 ) . The study confi rms the earlier reports which suggested that differentiated ( organized and redifferentiated ) cells and specialized organs generally produce most secondary products compared to dedifferentiated ( unorganized ) cells in cultures . "
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    ABSTRACT: Spilanthes acmella Murr., commonly known as “Toothache plant” and “Akarkara” of the family Asteraceae, holds an important place in Indian and global scenario owing to its medicinal properties. Different pharmacological experiments in a number of in vitro and in vivo models have convincingly demonstrated the ability of Spilanthes to exhibit anti-infl ammatory, antimalarial, antioxidant, antimicrobial and diuretic activities, lending support to the rationale behind several of its traditional uses. Spilanthes virtues are, to a large extent, attributable to its chemical constituents viz. alkylamides, phenolics, caumarin and triterpenoids. Of these, the most studied group has been the alkylamides, which are abundant in this plant and one which remains the most sought after by scientists is an antiseptic alkylamide, (2E, 6Z, 8E)-deca- 2,6,8-trienoic acid N-isobutyl amide, commonly known as spilanthol. Spilanthol has immense application in pharmaceuticals, food, health and body care products. The present review is an up-to-date and comprehensive analysis on in vitro conservation and biotechnological studies of Spilanthes. With this review authors aim to offer an experience background to researchers who intend to study on Spilanthes.
    Plant Reproductive Biology and Conservation, Edited by Rupam Kapoor, Inderdeep Kaur, Monika Koul, 02/2015: chapter An appraisal on in vitro conservation and biotechnological interventions in Spilanthes acmella Murr: pages 299-319; I.K. International, Delhi., ISBN: 9789382332909
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    • "This is the reason that makes researchers interested in developing methodologies to extract it from the plant and use reference standards, as dodeca-2(E),4(E)-dienoic acid isobutylamide for example, or in the comparison for identifying the peak of the compound in chromatographic analysis. However, the literature shows extraction taking 8-12 h using processes with methanol, ethanol, etc. (Dias et al., 2012; Singh and Chaturvedi, 2012). Processes assisted by microwaves can offer good advantages due to the volumetric heating of the material, reduced processing time and energy savings, with consequent high product quality. "
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    ABSTRACT: Jambu (Spilanthes oleracea) is a very popular plant in the Northern region of Brazil. It is an herbaceous plant belonging to Compositae family. The green leaves and stems are used in the Brazilian cuisine and the peculiar unique flavour is its main characteristics. Nowadays, many researchers and Brazilian companies are very interested in all parts of this plant due to its therapeutic potential and the use of its essential oil in cosmetic products. Instead of all of the components of the plant, the methodology in which one would obtain the amide spilanthol will be of interest to industry, since it is the principal compound in the essential oil. The main objective of this work was to obtain the proper calibration curve for the spilanthol from jambu flowers (area x concentration) using microwave extraction with ethanol and hexane (3:7) as solvent. The essential oil was extracted at 50 °C during 30 min and analysed in gas chromatography which confirmed the presence of spilanthol. The calibration curve was determined using concentrations of 0.05, 0.1, 0.25, 0.5 and 1.0 mg/mL in triplicates. The results showed a fast extraction method to remove efficiently the spilanthol from the flowers and a linear math model was arrived at with an excellent coefficient of determination of 99.8 %. The calibration model was used in later investigations of microwave extraction methods by the team.
    07/2013; 32:1783-1788. DOI:10.3303/CET1332298
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