Journal of Applied Pharmaceutical Science Vol. 9(09), pp 111-118, September, 2019 Available online at http://www.japsonline.com DOI: 10.7324/JAPS.2019.90916 ISSN 2231-3354 Tobacco resinoid (Nicotiana tabacum L.) as an active ingredient of cosmetic gels Venelina Popova1*, Yulian Tumbarski2, Tanya Ivanova1, Raina Hadjikinova1, Albena Stoyanova1 1Department of Tobacco, Sugar, Vegetable and Essential Oils, University of Food Technologies, Plovdiv, Bulgaria. 2Department of Microbiology, University of Food Technologies, Plovdiv, Bulgaria. ARTICLE INFO ABSTRACT Received on: 14/05/2019 Tobacco (Nicotiana tabacum L.) is not just an important industrial crop but also a source of phytochemicals with Accepted on: 15/07/2019 diverse biological and pharmacological activities. The study aimed at the investigation of the chemical composition Available online: 01/09/2019 and the antimicrobial activity of resinoids obtained from three types of tobacco grown in Bulgaria (Virginia, Burley, and Oriental), and the properties of a model cosmetic gel incorporating tobacco resinoid as an active ingredient. The GC-MS composition of the resinoids was presented by aliphatic hydrocarbons (56.54%–68.30%), oxygenated Key words: aliphatics (10.41%–20.45%), diterpenes (10.02%–18.63%), and triterpenes (4.55%–6.92%). The tested resinoids were Tobacco leaves, resinoid, more effective against Gram-positive bacteria (Bacillus subtilis ATCC 6633 and Bacillus cereus) and yeasts (Candida cosmetic gel, rheology, albicans NBIMCC 74) than against Gram-negative bacteria (Klebsiella pneumoniae). The resinoid of Oriental tobacco antimicrobial activity. was included as an active ingredient (1%) in a model cosmetic gel formulation. The rheological behavior of the gel characterized it as a non-Newtonian fluid. The inclusion of tobacco resinoid reduced gel viscosity in the applied stress range (1.5 ÷ 72.9 s-1). The outcomes from the study suggest that tobacco resinoid is a suitable ingredient of cosmetic gels, contributing to their organoleptic and functional properties. INTRODUCTION classes of bioactive chemical compounds, such as alkaloids, Tobacco (Nicotiana tabacum L. and to a lesser extent terpenoids (mono-, sesqui-, di-, and triterpenoids), phenolics N. rustica L.) has been long recognized as one of the most (phenolic acids, flavonoids, and coumarins), isoprenoids, and many economically important industrial crops and is cultivated in many others (Banožić et al., 2019; Rodgman and Perfetti, 2013). The countries around the world. Parallel to the commercial interest in chemistry and pharmacology of tobacco metabolites substantiate tobacco as leaf material for different tobacco products, the plant a reasonable interest in more innovative or broader use of the plant has been undeniably distinguished for its potential to synthesize through extraction and concentration of fractions with potential a plethora of secondary (or specialized) metabolites with various beneficial effects. On the other hand, tobacco has been historically biological or pharmacological activities. In a recent review, Jassbi regarded as a genuine essential oil-bearing (or aromatic) plant and et al. (2017) summarized that “cultivated tobacco (N. tabacum) has been used for obtaining aromatic products for the fragrance is among the most chemically and biologically well-studied industry. In fact, as Mookherjee and Wilson (1990) state, “no species of the plant kingdom, with more than 2500 characterized natural product in the flavor and fragrance industry can match metabolites, updated by continuous research”. These metabolites, tobacco for the number of volatile constituents which have found in high concentrations in tobacco leaves, include many been identified”. Traditional (or established) aromatic products obtained from tobacco include tobacco concrete (by extraction with nonpolar solvents, like n-hexane), tobacco resinoid (by extraction with polar solvents, like ethanol), and tobacco absolute (by re-extraction of concrete or resinoid with polar solvents at low *Corresponding Author Venelina Popova, Department of Tobacco, Sugar, Vegetable and Essential temperatures) (Baser and Buchbauer, 2010; Bauer et al., 2001), Oils, University of Food Technologies, Plovdiv, Bulgaria. which are used in perfumery or in the casing and top-flavoring of E-mail: vpopova2000 @ abv.bg tobacco blends due to their specific olfactory profiles. Additionally, © 2019 Venelina Popova et al. This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/). 112 Popova et al. / Journal of Applied Pharmaceutical Science 9 (09); 2019: 111-118 these extracts have demonstrated antimicrobial, antioxidant, and trifluoroacetamide (BSTFA, Supelco) and incubated at 70°C for 45 pharmacological properties (Docheva et al., 2018; Ru et al., 2012; minutes. The sample was then diluted with 150 µl chloroform and Vijayabhaskar et al., 2014) and could be considered as active injected (1 µl volume) in a system comprised an Agilent 7890A ingredients (natural supplements or antioxidants) in cosmetic gas chromatograph (Agilent Technologies) and mass selective preparations. The growing interest in such natural products and the detector Agilent 5975C (Agilent Technologies). The column was inclusion of various plant extracts and phytochemicals in cosmetic HP-5ms (30 m × 0.32 mm; film thickness 0.25 µm) operated products (Fedorovska et al., 2018; Iswandana et al., 2018; Khor under the following conditions: temperature increase from 40°С et al., 2018) substantiate comprehensive studies of the (0 minute) to 230°С at 5°С/minute (held at 230°С for 10 minutes); composition and properties of extraction products derived from injector and detector temperature 250°С; carrier gas helium at 1 tobacco representing different genotypes and regions. Due to the ml/minute constant flow; MSD scan range m/z = 50–550; split economic importance of tobacco, the species has developed into mode (5:1). numerous commercial types and varieties, each with characteristic properties, composition, and use (Dyulgerski and Docheva, 2017; Antimicrobial activity Korubin-Aleksoska and Ahmad, 2016; Radoukova and Dyulgerski, Test microorganisms 2018). In general, three major types of tobacco are produced and Twelve microorganisms including Gram-positive used in cigarettes: flue-cured Virginia, Oriental (also known as bacteria (B. subtilis ATCC 6633, Bacillus cereus—clinical isolate, aromatic or Turkish tobacco, often labeled as the richest in flavor Staphylococcus epidermidis ATCC 12228, Staphylococcus aureus among all tobacco types), and Burley light air-cured. Considering ATCC 25923, Listeria monocytogenes ATCC 19111, Enterococcus the historically established traditions in the large-scale production faecalis—clinical isolate), Gram-negative bacteria (Salmonella of all of the three types in Bulgaria, as well as the availability of the enteritidis—clinical isolate, Klebsiella pneumoniae—clinical raw material and the respective processing facilities and network, isolate, Escherichia coli ATCC 25922, Proteus vulgaris ATCC we hypothesized that the leaves from the three types of tobacco 6380, Pseudomonas aeruginosa ATCC 9027), and yeasts (Candida grown in the country could be processed to aromatic products albicans NBIMCC 74) from the collection of the Department of with specific composition and character. We further hypothesized Microbiology at the University of Food Technologies, Plovdiv, that tobacco resinoids would reveal antimicrobial activity and Bulgaria, were selected for the antimicrobial test. characteristic olfactory profile and could be included in cosmetic preparations (creams, gels, etc.) as an active ingredient. Culture media Therefore, the aim of the present study was to obtain Luria–Bertani glucose agar (LBG-agar) medium and characterize (composition by gas chromatography-mass (Laboratorios Conda S.A., Madrid, Spain) containing tryptone, spectrometry (GC-MS), antimicrobial activity) the resinoid yeast extract, NaCl, glucose, and agar was used for cultivation of from the leaves of different tobacco types and the properties of a test microorganisms, as well as for implementation of agar well cosmetic gel containing tobacco resinoid as an active ingredient. diffusion assay. For these two purposes, 50 g of LBG-agar medium MATERIAL AND METHODS was dissolved in 1 l of deionized water, the final pH was adjusted to 7.5 and then autoclaved at 121°C for 20 minutes before use. Plant material Antimicrobial assay The plant material were cured and aged (for 12 months) tobacco leaves, provided by local leaf processing factories, which The antimicrobial activity of tobacco resinoids was were further sorted to obtain uniform samples. Three tobacco types determined by the standard agar well diffusion method in were studied, all representing local varieties grown in South Bulgaria LBG-agar medium (Tumbarski et al., 2018). The pathogenic tobacco-producing region—flue-cured Virginia (FCV), Burley (BU), bacteria and yeasts were cultured on LBG-agar medium at 37ºC and Oriental (OR). The leaves were oven-dried (40°C; 6 hours), for 24 hours, while B. subtilis ATCC 6633 and B. cereus—at 30ºC ground, and sieved to particle size required by the respective analysis. for 24 hours. The inocula of test microorganisms were prepared by The moisture content of the leaves before processing was 7.21% ± homogenization of a small amount of biomass in 5 mL of sterile 0.05% (FCV), 7.93% ± 0.06% (BU), and 7.56% ± 0.05%