CHEMICAL COMPOSITION OF VOLATILE OIL OF NIGELLA SATIVA SEEDS

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1 WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES SJIF Impact Factor Volume 3, Issue 10, Research Article ISSN CHEMICAL COMPOSITION OF VOLATILE OIL OF NIGELLA SATIVA SEEDS Rabindra Kumar Singh 1*, Kamal Nayan Jhunjhunwalla 2 1 Research & Development Manager, M/S AFF Aromatics Pvt. Ltd, Howrah, India. 2 Research Associate, M/S AFF Aromatics Pvt. Ltd, Howrah, India. Article Received on 18 August 2014, Revised on 10 September 2014, Accepted on 02October 2014 *Correspondence for Author Dr.Rabindra Kumar Singh Research & Development Manager, M/S AFF Aromatics Pvt. Ltd, Howrah, India. ABSTRACT The composition of the essential oil of Nigella sativa obtained through hydro-distillation. The compositional analysis carried out by GC and GCMS, which identify 35 components representing 97.09% of total composition of oil. The major components are para-cymene (53.6%), alpha Thujene (15.18%), beta pinene (3.7%) and alpha pinene (3.3%). This mix of monoterpenes imparts its odour a strong terpene-like top note. KEYWORDS: Nigella Sativa, Blackseed, Para cymene, Thymoquinone, Essential oil, Black seed. INTRODUCTION The species Nigella Sativa belonging order ranunculales family ranunculaceae. Which is also known as blackseed, kala jeera or mangrail. It is used for culinary purposes. However, blackseed oil has penetrated boundaries to enter the aromatherapy market due to its wide-ranging health benefits (Paarakh et al 2010) (Salem 2005). Nigella sativa occupies a major place in the cultures of south East Asia and south Asia. It serves a wide range of medicinal purposes, which is rendering it increasingly important in the medical world (Bourgou et al 2008). In addition, it is an integral part of the cuisine of South Asia and South East Asia, used as a flavour for traditional breads. There is a notion that the Islamic prophet Muhammad called black seed a major medicinal species. The ant cancerous effects of thymoquinone have been studied. The potential of thymoquinone and its derivative molecules as anticancer agents has been established (Woo2011, Molecules2006). Its potential against cytotoxic activity has also been established (Harzallah et al 2011). The allelopathic potential of this species has recently been outlined (Zribi et al Vol 3, Issue 10,

2 2014). It is used as an anti-diabetic, anti-hepato-toxic, anti-cancerous agent, antibacterial, antifungal, and anti-parasitic and as a therapeutic. Thymoquinone, a major component of the oil also works as an anti-epileptic (Kamil et al 2013). In addition to these uses, blackseed is an integral part of South Asian and South West Asian cuisines. MATERIALS AND METHODS Raw Material: The black seed used for this experiment were collected from Nadia district, West Bengal. In the local region, this species is known as kala jeera. Experimental Design: Fresh Nigella sativa seeds were hydro distilled in three batches (2kg seeds with 6 litres of water each) in a Clevenger apparatus for 4-5 hours at approximately 80 C. The distillate was extracted with hexane. The distillate was extracted with hexane (5x10ml) each batch, after that the hexane layer was dried over anhydrous sodium sulphate and hexane distilled off on a gently heated water bath at 40 C. Total oil obtained after distilled off the hexane is grams. The yield of the oil (W/W) was thus calculated to be 0.48 %. GC-MS Analysis The oil analysis was performed using an Agilent 7890a series GC connected with 5975C MSD and FID connected with splitter with following conditions mention in Table 1. Table 1: GC & GCMS Parameters. Column DB5-MS Column dimensions 60.0m x 0.25mm x 0.25 µm Carrier Gas Helium Injector Temperature 270 Detector Temperature 270 MS mode Scan mode ( amu) Constant Pressure 23psi Ionization Electron Ionization at 70 ev Split ratio 1:50 Oven conditions Rate (C/min) Value Hold time Run time ( C ) ( min) (min) Initial Ramp Vol 3, Issue 10,

3 After analysis the identification of component was carried out by mass fragmentation pattern matching with NIST, Willey and Adam s mass library. The identification has done by GCMS while quantification done by FID. RESULTS AND DISCUSSION Monoterpenes constitute a large fraction of the total composition of the volatile oil of West Bengal s Nigella Sativa seeds. Seven monoterpenes alpha Thujene, alpha Pinene, sabinene, beta pinene, para Cymene, Limonene and gamma terpinene together constitute above 81% of the total composition of the oil. The terpenoids in this oil include terpene alcohols, terpene ketones and terpene acetates. The major chiral terpenes in this molecule are alpha pinene and limonene, which constitute above 5% of the total oil. Terpenoids carvacrol, linalool, and terpin-4-ol, together constitutes 6.44% of the oil. The compositions of component shown in Table 2 while the GC graph and margin of error given in Graph1 and 2. Figures and Tables Figure 1: GC Graph of Nigella Sativa Seed Oil Vol 3, Issue 10,

4 Figure 2: The major components of the hydro-distilled oil with the respective margins of error Table 2: Composition of nigella sativa seed oil COMPONENT RT EXP 1 EXP. 2 EXP. 3 MEAN MARGIN OF ERROR Alpha Thujene Alpha Pinene Camphene Sabinene Beta Pinene Beta Myrcene Alpha Phellandrene Alpha Terpinene Para Cymene DL-Limonene Beta Phellandrene Eucalyptol Gamma Terpinene Alpha Terpinolene Styrol Linalool Camphor Terpin-4-ol Vol 3, Issue 10,

5 P-Cymen-8-ol Alpha Terpineol Carvone Thymoquinone Thujyl Acetate Bornyl Acetate Carvacrol Alpha Longipinene Alpha Ylangene Longicyclene Longifolene Undecanone Beta Bisabolene Trans- Cadinadiene Undecanal Ambrettolide Pimaradiene CONCLUSION The volatile oil of a certain variety of Iranian blackseed been reported to contain 38.3% transanethole (a phenylpropene). In addition, 46.1% of the total oil composition belongs to phenylpropenes and 26% belongs to monoterpenoid hydrocarbons (Nickavar2003). This significantly differs from the composition obtained in this study - 81% monoterpenes. In addition, the composition also significantly varies from that of the Moroccan variety of Nigella sativa oil (Gharby et al 2013). Thymoquinone - a major component of Nigella sativa essential oil extracted from across a variety of sources is present in a very low quantity in this essential oil. Different extraction methods should be attempted on the same raw material to increase the quantity of thymoquinone in the oil. As an example, the Soxhlet extraction of Nigella Sativa seeds followed by the distillation of the volatile oil has been performed to achieve greater percentage of thymoquinone. This difference could have been caused due to either: a) The geographical difference between the crop of Iran and West Bengal b)the methodical difference between the processes. (Soxhlet extraction followed by distillation v distillation) (Kiralan et al 2014) c) Sowing date, sowing rate and location (Ullah et al 2013). Vol 3, Issue 10,

6 ACKNOWLEDGEMENTS I would like to extend my gratitude to our R&D team members of AFF Aromatics Pvt Ltd. REFERENCES 1. Paarakh PM. Nigella sativa Linn. A comprehensive review. Indian Journal of Natural Products and Resources, 2010; 1(4): Salem ML. Immunomodulatory and therapeutic properties of the Nigella sativa L. seed. International Immunopharmacology, 2005; 5(13-14): Bourgou S, Ksouri R, Bellila A, Skandrani I, Falleh H, & Marzouk B. Phenolic composition and biological activities of Tunisian Nigella sativa L. shoots and roots. Comptes Rendus Biologies, 2008; 331(1): Woo CC, Loo SY, Gee V, Yap CW, Sethi G, Kumar AP & Tan KHB. Anticancer activity of thymoquinone in breast cancer cells: possible involvement of PPAR-γ pathway. Biochemical Pharmacology, 2011; 82(5): Molecules L, Products N, & All EBV. The medicinal potential of black seed ( Nigella sativa ) and its components. Asian Journal of Physical Sciences, 2006; Jrah Harzallah H, Kouidhi B, Flamini G, Bakhrouf A, & Mahjoub T. Chemical composition, antimicrobial potential against cariogenic bacteria and cytotoxic activity of Tunisian Nigella sativa essential oil and thymoquinone. Food Chemistry, 2011; 129(4): Zribi I, Omezzine F & Haouala R. Variation in phytochemical constituents and allelopathic potential of Nigella sativa with developmental stages. South African Journal of Botany, 2014; 94: Kamil, ZH, Spectacular Black Seeds ( Nigella sativa ) Medical Importance Review, Medical Journal of Babylon. 2013; 10(4): Nickavar B, Mojab F, Javidnia K, & Amoli MAR. Chemical composition of the fixed and volatile oils of Nigella sativa L. from Iran Zeitschrift Für Naturforschung. Journal of Biosciences, 2003; 58(9-10): Gharby S, Harhar H, Guillaume D, Roudani A, Boulbaroud S, Ibrahimi M, Charrouf Z. Chemical investigation of Nigella sativa L. seed oil produced in Morocco, Journal of the Saudi Society of Agricultural Sciences, Kiralan, M, Özkan G, Bayrak A, & Ramadan MF. Physicochemical properties and stability of black cumin (Nigella sativa) seed oil as affected by different extraction methods, Industrial Crops and Products, 2014; 57: Vol 3, Issue 10,

7 12. Ullah H & Honermeier B. Fruit yield, essential oil concentration and composition of three anise cultivars (Pimpinella anisum L.) in relation to sowing date, sowing rate and locations. Industrial Crops and Products, 2013; 42: Vol 3, Issue 10,