Citrus oils composition, advanced analytical techniques, contaminants, and biological activity

Citrus Oils Composition, Advanced Analytical Techniques, Contaminants, and Biological Activity K10034_C000.indd i 9/21/2010 8:32:24 PM Medicinal and Aromatic Plants — Industrial Profiles Individual volumes in this series provide both industry and academia with in-depth coverage of one major genus of industrial importance Series Edited by Dr Roland Hardman Volume Valerian, edited by Peter J Houghton Volume Perilla, edited by He-ci Yu,   Kenichi Kosuna and Megumi Haga Volume Poppy, edited by Jenö Bernáth Volume Cannabis, edited by David T Brown Volume Neem, edited by H.S Puri Volume Ergot, edited by Vladimír Kˇren and   Ladislav Cvak Volume Caraway, edited by Éva Németh Volume Saffron, edited by Moshe Negbi Volume Tea Tree, edited by Ian Southwell and   Robert Lowe Volume 10 Basil, edited by Raimo Hiltunen and   Yvonne Holm Volume 11 Fenugreek, edited by   Georgios Petropoulos Volume 12 Ginkgo biloba, edited by   Teris A Van Beek Volume 13 Black Pepper, edited by P.N Ravindran Volume 14 Sage, edited by Spiridon E Kintzios Volume 15 Ginseng, edited by W.E Court K10034_C000.indd ii Volume 16 Mistletoe, edited by Arndt Büssing Volume 17 Tea, edited by Yong-su Zhen Volume 18 Artemisia, edited by Colin W Wright Volume 19 Stevia, edited by A Douglas Kinghorn Volume 20 Vetiveria, edited by Massimo Maffei Volume 21 Narcissus and Daffodil, edited by   Gordon R Hanks Volume 22 Eucalyptus, edited by John J.W Coppen Volume 23 Pueraria, edited by Wing Ming Keung Volume 24 Thyme, edited by E Stahl-Biskup and   F Sáez Volume 25 Oregano, edited by Spiridon E Kintzios Volume 26 Citrus, edited by Giovanni Dugo and   Angelo Di Giacomo Volume 27 Geranium and Pelargonium, edited by   Maria Lis-Balchin Volume 28 Magnolia, edited by Satyajit D Sarker and Yuji Maruyama Volume 29 Lavender, edited by Maria Lis-Balchin Volume 30 Cardamom, edited by P.N Ravindran and K.J Madhusoodanan 9/21/2010 8:32:25 PM Volume 31 Hypericum, edited by Edzard Ernst Volume 32 Taxus, edited by H Itokawa and   K.H Lee Volume 33 Capsicum, edited by Amit Krish De Volume 34 Flax, edited by Alister Muir and   Niel Westcott Volume 35 Urtica, edited by Gulsel Kavalali Volume 36 Cinnamon and Cassia, edited by   P.N Ravindran, K Nirmal Babu   and M Shylaja Volume 37 Kava, edited by Yadhu N Singh Volume 38 Aloes, edited by Tom Reynolds Volume 39 Echinacea, edited by   Sandra Carol Miller   Assistant Editor: He-ci Yu Volume 40 Illicium, Pimpinella and Foeniculum,      edited by Manuel Miró Jodral Volume 41 Ginger, edited by P.N Ravindran   and K Nirmal Babu Volume 42 Chamomile: Industrial Profiles,   edited by Rolf Franke and   Heinz Schilcher Volume 43 Pomegranates: Ancient Roots to   Modern Medicine, edited by   Navindra P Seeram,   Risa N Schulman and David Heber K10034_C000.indd iii Volume 44 Mint, edited by Brian M Lawrence Volume 45 Turmeric, edited by P N Ravindran,   K Nirmal Babu, and K Sivaraman Volume 46 Essential Oil-Bearing Grasses, edited by Anand Akhila Volume 47 Vanilla, edited by Eric Odoux   and Michel Grisoni Volume 48 Sesame, edited by Dorothea Bedigian Volume 49 Citrus Oils, edited by Giovanni Dugo and Luigi Mondello 9/21/2010 8:32:26 PM K10034_C000.indd iv 9/21/2010 8:32:26 PM Citrus Oils Composition, Advanced Analytical Techniques, Contaminants, and Biological Activity Edited by Giovanni Dugo and Luigi Mondello Medicinal and Aromatic Plants — Industrial Profiles K10034_C000.indd v 9/21/2010 8:32:26 PM CRC Press Taylor & Francis Group 6000 Broken Sound Parkway NW, Suite 300 Boca Raton, FL 33487-2742 © 2011 by Taylor and Francis Group, LLC CRC Press is an imprint of Taylor & Francis Group, an Informa business No claim to original U.S Government works Printed in the United States of America on acid-free paper 10 International Standard Book Number: 978-1-4398-0028-7 (Hardback) This book contains information obtained from authentic and highly regarded sources Reasonable efforts have been made to publish reliable data and information, but the author and publisher 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Drive, Danvers, MA 01923, 978-750-8400 CCC is a not-for-profit organization that provides licenses and registration for a variety of users For organizations that have been granted a photocopy license by the CCC, a separate system of payment has been arranged Trademark Notice: Product or corporate names may be trademarks or registered trademarks, and are used only for identification and explanation without intent to infringe Library of Congress Cataloging-in-Publication Data Citrus oils : composition, advanced analytical techniques, contaminants, and biological activity / edited by Giovanni Dugo and Luigi Mondello p cm (Medicinal and aromatic plants industrial profiles ; 49) Includes bibliographical references and index ISBN 978-1-4398-0028-7 Citrus oils Composition Citrus oils Analysis Citrus oils Therapeutic use I Dugo, Giovanni II Mondello, Luigi III Title: Composition, analysis, contamination and properties of citrus oils IV Series: Medicinal and aromatic plants industrial profiles ; v 49 TP959.C54C575 2011 665’.3 dc22 2010012530 Visit the Taylor & Francis Web site at http://www.taylorandfrancis.com and the CRC Press Web site at http://www.crcpress.com K10034_C000.indd vi 9/21/2010 8:32:26 PM To the wonderful women of my family To my grandmother, Francesca, who cheered and protected me To my mother, Paola, who was always excessively proud of my limited success To my wife, Anna, who loved and assisted me, always indulgent and understanding To my daughters, Paola, Monica, and Laura, who showed to me how lucky a father can be To the sweetest daughters of my daughters, Alice, Viola, and Laura, who, more than anything else, give a sense to my life and sweeten my old age Giovanni Dugo To my wife, Paola, and to my children, Alice and Viola, for their understanding and patience while I spent seemingly endless evenings and weekends working in my research laboratory To my parents for their love and for believing in me and encouraging me in my career Luigi Mondello K10034_C000.indd vii 9/21/2010 8:32:26 PM K10034_C000.indd viii 9/21/2010 8:32:26 PM Contents Series Preface xi Preface xiii Editors xv Contributors xvii Chapter Composition of the Volatile Fraction of Citrus Peel Oils Giovanni Dugo, Antonella Cotroneo, Ivana Bonaccorsi, Alessandra Trozzi Chapter Volatile Components in Less Common Citrus Species 163 Estelle Delort, Regula Naef Chapter Composition of Distilled Oils 193 Luis Haro-Guzmán Chapter Concentrated Citrus Oils 219 Herta Ziegler Chapter Composition of Petitgrain Oils 253 Giovanni Dugo, Antonella Cotroneo, Ivana Bonaccorsi Chapter Extracts from the Bitter Orange Flowers (Citrus aurantium L.): Composition and Adulteration 333 Giovanni Dugo, Louis Peyron, Ivana Bonaccorsi Chapter The Chiral Compound of Citrus Oils 349 Luigi Mondello, Rosaria Costa, Danilo Sciarrone, Giovanni Dugo Chapter The Oxygen Heterocyclic Components of Citrus Essential Oils .405 Paola Dugo, Marina Russo Chapter Carotenoids of Citrus Oils 445 Paola Dugo, Daniele Giuffrida Chapter 10 Minor Components in Extracts of Citrus Fruits 463 Regula Naef ix K10034_C000.indd ix 9/21/2010 8:32:26 PM Biological Activities of Citrus Essential Oils 535 Finally, lemon pure essential oils were shown able to inhibit heat shock–induced apoptosis in the human astrocyte cell line CCF-STTG1 and in primary cultured rat astrocytes, by blocking caspase-3 activation, DNA fragmentation and poly-ADP-ribose polymerase fragmentation (Koo et al., 2002) 13.3.2 ANXIOLYTIC AND SEDATIVE PROPERTIES The word “aromatherapy” combines two words: aroma (a fragrance or sweet smell) and therapy (a treatment) Aroma and massage therapy are the practice of using essential oils for psychological and physical well-being via inhalation or massage The term “aromatherapy” is used to describe a wide range of practices involving odorous substances, although only aroma delivery through inhalation for inducing psychological or physical effects can be correctly defined as aromatherapy Nevertheless, the clinical use of essential oils and their volatile constituents via inhalation or massage has expanded worldwide (Edris, 2007) A number of essential oils are currently in use as aromatherapy agents to relieve anxiety, stress, and depression Popular anxiolytic oils include those from Citrus sinensis, Citrus aurantium, and Citrus limon, suggesting central nervous system action The anxiolytic, anticonvulsant, and sedative effects following oral administration of C aurantium L essential oil have been characterized in different behavioral models in mice (Carvalho-Freitas and Costa, 2002; Pultrini et al., 2006); these beneficial effects are not accompanied by deficits in general activity or motor coordination Furthermore Komiya et al (2006) and Ceccarelli et al (2004) have demonstrated the antistress action of the essential oil of lemon by means of different behavioral models in mice and rats; these anxiolytic, antidepressant-like effects appeared to be mediated via suppression of dopaminergic activity related to enhanced serotoninergic neurons As to studies carried out in humans, Lehrner et al (2000) reported that exposure to ambient odor of orange diffused in a dental office has a relaxant effect; in particular, women exposed to orange odor had a lower level of state anxiety, a more positive mood and a higher level of calmness These data support the other evidences of sedative properties of the natural essential oil of orange Conversely, no positive effect on anxiety levels of adults accompanying children to a pediatric emergency department was observed by Holm and Fitzmaurice (2008) when aromatherapy alone (neroli essential oil) or music in addition to aromatherapy diffused in the waiting area These different results could be because of environmental conditions or application modalities of the aromatherapy 13.3.3 MODULATION OF NEUROTRANSMITTER FUNCTIONS A large number of biological effects elicited by exposure to citrus essential oils may be related to their capability to modulate neurotransmitter functions BEO contains into its volatile fraction some monoterpene hydrocarbons able to stimulate glutamate release by transporter reversal and/or by exocytosis, depending on the dose administered (Morrone et al., 2007) In fact intraperitoneal administration of BEO in rats could significantly affect the extracellular concentration of aspartate, glycine, and taurine; furthermore, when perfused into the hippocampus, BEO produced a significant increase of extracellular aspartate, glycine, and taurine as well as of GABA and glutamate These effects appeared to be dependent on the glutamate transporter blocker dl-threo-β-benzyloxyaspartic acid and on extracellular Ca2+ More recently Rombolà et al (2009) have described the systemic effects of this phytocomplex on gross behavior and EEG activity recorded from the hippocampus and cerebral cortex of the rat Systemic administration of BEO produces dose-dependent increases in locomotor and exploratory activity that correlate with significant changes in the EEG spectrum Fukumoto et al (2006) used brain-tissue slices to demonstrate the capability of R-limonene, γ-terpinene, and citral, major components of lemon essential oil, and of their metabolites on K10034_C013.indd 535 9/21/2010 5:53:26 PM 536 Citrus Oils monoamine release; interestingly, the metabolites of these monoterpenes have a stronger effect on monoamine release from brain tissue than the monoterpene compounds themselves The antidementia effects of (S)-(–)-limonene and (S)-(–)-perillyl alcohol have been reported by Zhou et al (2009) In fact, these compounds showed strong ability to improve memory impaired by scopolamine in experimental animals; brain dopamine concentration of the scopolamine group was significantly lower than that of the control group, but this phenomenon was reversed by pretreatment with (S)-(–)-limonene or (S)-(–)-perillyl alcohol Furthermore, these two lemon essential oil components could inhibit acetylcholinesterase (AChE) activity in vitro Besides the lemon essential oil, the essential oils of Citrus paradisi (pink grapefruit) were also shown able to inhibit AChE activity, very likely because of the nootkatone and auraptene contained in them (Miyazawa et al., 2001) Today there is a raising interest in studying the alterations on nervous system functions following inhalation of citrus essential oils For example, essential oil from citrus lemon, inhaled by female rats experiencing a persistent nociceptive input, can affect and modulate the increase in hippocampal acetylcholine release induced by pain (Ceccarelli et al., 2002); however, this effect appeared to be gender dependent because it was evident only in female, and not in male, animals Consistent with these findings, long-term exposure of rats to lemon essential oil can induce significant, at times sex-specific, changes in neuronal circuits involved in anxiety and pain (Ceccarelli et al., 2004) In experimental animals, olfactory stimulation with scent of grapefruit oil (SGFO) enhances sympathetic nerve activities, suppresses gastric vagal nerve activity (GVNA) and increases plasma glycerol concentration; furthermore olfactory stimulation with SGFO or with limonene elevates renal sympathetic nerve activity (RSNA) and blood pressure and lowers GVNA in urethane-anesthetized rats (Tanida et al., 2005) The authors suggested that SGFO and its active component limonene affect autonomic neurotransmission and blood pressure through central histaminergic nerves and the suprachiasmatic nucleus In agreement with data from experimental animals, fragrance inhalation of grapefruit oil in normal humans results in an increase of sympathetic activity (Haze et al., 2002) The activation of sympathetic nerve innervating the white adipose tissue is known to facilitate lipolysis, resulting in a suppression of body weight gain Olfactory stimulation with SGFO excites the sympathetic nerve innervating the white and brown adipose tissue and adrenal gland and inhibits the parasympathetic gastric nerve in rats (Shen et al., 2005) Limonene induces responses similar to those caused by SGFO The capability of SGFO, and particularly of its primary component limonene, to affect autonomic nerves and enhance lipolysis is mediated through a histaminergic response An increased sympathetic nerve activity to white adipose tissue in anesthetized rat was shown by Niijima and Nagai (2003) also following olfactory stimulation with the scent of lemon oil Finally, one has to mention that the essential oils of hakyul (Citrus natsudaidai Hayata), yuza (Citrus junos Sieb ex Tanaka), and lemon have a significant lipolytic effect, as shown by Choi (2006) using an olive oil model solution Among the authentic compounds relating to citrus-peel oils, octanal, γ-terpinene, limonene, terpinen-4-ol, nerol, p-cymene and geranyl acetate showed the highest lipolytic effect; thus monoterpene hydrocarbons having one or two double bonds would have stronger lipolytic effect than those having three double bonds 13.4 ANTINOCICEPTIVE PROPERTIES OF CITRUS ESSENTIAL OILS Due to the increasing use of aromatherapy oils, the antinociceptive properties of citrus essential oils have been investigated by several authors Sakurada et al (2009) reported that the capsaicin-induced nociceptive response in mice is significantly reduced by intraplantar injection of BEO, while sweet orange essential oil is without effect Among the monoterpene hydrocarbons found in BEO volatile fraction, linalol might be responsible for the antinociceptive effects of this phytocomplex In fact, linalol possesses antinociceptive, antihyperalgesic, and anti-inflammatory properties in different experimental animals Linalol effects K10034_C013.indd 536 9/21/2010 5:53:26 PM Biological Activities of Citrus Essential Oils 537 in neuropathic pain have been investigated by Berliocchi et al (2009) Chronic administration of linalol is able to reduce mechanical allodynia (but not sensitivity to noxious radiant heat) following spinal nerve ligation as model of neuropathic pain in mice Mechanisms other than an action on inflammatory processes may be supposed to mediate the protective effect of linalol in this model of neuropathic pain Also the polymethoxylated flavone 3′,4′,3,5,6,7,8-heptamethoxyflavone (HMF), found in citrus essential oils, possesses anti-inflammatory properties In fact, intraperitoneal (but not oral) administration of HMF proved to induce an anti-inflammatory effect when studied in the bacterial lipopolysaccharide-challenge/tumor necrosis factor-α (TNF-α) response in mice and in the carrageenan/paw edema assay in rats (Manthey and Bendele, 2008) This effect appeared to be related to the different bioavailability of the intact HMF following oral and intraperitoneal administration, while the glucuronidated HMF metabolites seem to be inactive There is evidence about the ability of citrus essential oils to modulate (not only behavioral but also neuronal) responses related to nociception and pain also following inhalation and aromamassage therapy Lemon essential oil inhaled in rats experiencing a persistent nociceptive input decreased behavioral responses related to the nociceptive stimulus, influencing c-Fos expression (used to test the degree of neuronal activation of areas belonging to the limbic system) in the arcuate and paraventricular nuclei of the hypothalamus and in the dentate gyrus of the hippocampus (Aloisi et al., 2002), with an effect dependent on animal gender Furthermore, the aroma-massage therapy with citrus essential oils might have potential as an alternative/complementary method for short-term pain relief, as shown by Yip and Tam (2008), who carried out a double-blind, placebo-controlled study to assess the efficacy of the aroma massage (6 massage sessions for weeks) with aromatic essential oils (Zingiber officinalis and Citrus sinesis) on older subjects with moderate to severe knee pain Specific formulations can improve the anti-inflammatory efficacy of citrus essential oils For example the combination of Dead Sea magnesium chloride (MgCl2) and citrus oils, tested in a subcutaneous chamber model in mice, resulted in lower levels of TNF-α and leukocyte migration without changes in IL-10 levels, in comparison with controls (Mizrahi et al., 2006) One has to point out that, despite the anti-inflammatory effects, citrus essential oils not affect prostaglandin basal levels In fact, the essential oil from Citrus aurantium (that is used in traditional medicine to treat gastritis and gastric disorders) and its main component limonene provide, when orally administered in rats, effective gastroprotection against lesions induced by absolute ethanol and nonsteroidal anti-inflammatory drugs, increasing gastric mucus production and without interfering with gastric H+ secretion, serum gastrin, or glutathione level in gastric mucosa (Moraes et al., 2009) 13.5 ANTIOXIDANT PROPERTIES OF CITRUS ESSENTIAL OILS Reactive oxygen species, as well as reactive nitrogen species, play either harmful or beneficial role in biological systems Beneficial effects of ROS include physiological roles in cellular responses against infectious agents and in several cellular signaling pathways Harmful effects are due to high concentrations of ROS, which can damage biomolecules, including lipids, proteins, and nucleic acids The harmful effects of ROS are counterbalanced by the antioxidant action of both antioxidant enzymes and nonenzymatic antioxidants; however, despite the presence of the cellular antioxidant system, oxidative damage accumulates during the life cycle and has been proposed to play a pivotal role in the development of age-dependent diseases such as atherosclerosis, arthritis, neurodegenerative disorders and cancer There is today increasing interest in the radical scavenging activities of some natural antioxidants, especially those found in edible plants, which might have a role in preventing various chronic pathologies and find applications in food and cosmetic industry Essential oils and some of their K10034_C013.indd 537 9/21/2010 5:53:26 PM 538 Citrus Oils components are highly lipophilic and have been shown to possess antioxidant properties However, little is known about the antioxidant properties of essential oils from edible plants and, in particular, of citrus essential oils Thirty-one kinds of citrus essential oils and their components were investigated for their radical scavenging properties against the stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical (Choi et al., 2000), and appeared to have an antioxidant power similar to that of Trolox; the citrus volatile components geraniol, terpinolene, and γ-terpinene showed marked scavenging activity against DPPH γ-Terpinene was also found to inhibit human low density lipoprotein (LDL) oxidation induced in vitro by Cu2+ or 2.2′-azobis(2-amidinopropane) dihydrochloride (AAPH; Edris, 2007) Also, essential oils from sweet orange peels have been investigated in terms of DPPH radical scavenging, β-carotene bleaching and nitrite scavenging activities (Anagnostopoulou et al., 2006) Misharina and Samusenko (2008) demonstrated the good antioxidant properties of essential oils from lemon (Citrus limon L.) and especially from pink grapefruit (Citrus paradisi L.) by capillary gas-liquid chromatography against the oxidation of the aliphatic aldehyde hexanal to the carboxylic acid Furthermore, the essential oil from Citrus karna Raf, containing d-limonene as major constituent, together with α-pinene and β-pinene as minor constituents, showed significant inhibition against the oxidation of linoleic acid in the β-carotene-linoleic acid system (Malhotra et al., 2009) The results indicate a main role for d-limonene in antioxidant activity Finally, bergaptol, a bioactive compound isolated from grapefruit peel oil, showed radical scavenging activity using 2,2′-azobis(3-ethylbenz-thiazoline-6-sulfonic acid) and DPPH (Girennavar et al., 2007); this compound is also a potent inhibitor of debenzylation activity of CYP3A4 enzyme Oxidation of LDL has been implicated in atherogenesis for several years Therefore many researchers are looking for potent antioxidants, which are able to inhibit LDL oxidation and thus lower the risk for atherosclerosis The antioxidative action of citrus essential oil components was studied on LDL oxidation in vitro (Grassmann et al., 2001; Takahashi et al., 2003) Lemon oil and, among its volatile compounds, γ-terpinene showed the strongest antioxidative effect and inhibited both copperand AAPH-induced oxidation of LDL In particular the loss of carotenoids during LDL oxidation appeared to be strongly retarded by lemon oil and γ-terpinene (Grassmann et al., 2001) Recently, it has been suggested that Lectin-like oxyLDL receptor-1 (LOX-1) is involved in smooth muscle cell (SMC) proliferation and neointima formation in injured blood vessels The BEO nonvolatile fraction has a protective effect on LOX-1 expression and free-radical generation in common carotid-artery injury induced by balloon angioplasty in rats (Mollace et al., 2008) These results suggest that natural antioxidants may be relevant in the treatment of vascular disorders in which proliferation of SMCs and oxyLDL-related endothelial cell dysfunction are involved Besides terpenes, other components found in citrus essential oils have well-established antioxidant properties For example, Tirkey et al (2005) demonstrated the good protective effect of the citrus bioflavonoid hesperidinin against liver and kidney oxidative damage induced in rats by CCl4 (a toxic agent which is metabolized to produce free radicals and widely used to experimentally induce acute and chronic hepatic and renal injuries in rodents) Limonene is used as flavoring agent in a wide range of food Since it is lipid soluble, limonene is often added to foods in oil-in-water emulsions, which are susceptible to both physical instability and oxidative degradation, leading to loss of aroma and formation of off-flavors Whey protein isolate (WPI) could inhibit the oxidative deterioration of limonene in oil-in-water emulsions (Djordjevic et al., 2008), through the formation of a cationic emulsion-droplet interface and/or the ability of amino acids in WPI to scavenge free radical and chelate pro-oxidative metals Also a sodium dodecyl sulfate-chitosan complex has appeared able to inhibit the oxidative deterioration of limonene because of the formation of a cationic and thick emulsion-droplet interface that could repel pro-oxidative metals (Djordjevic et al., 2007) K10034_C013.indd 538 9/21/2010 5:53:26 PM Biological Activities of Citrus Essential Oils 13.6 539 CHEMOPREVENTIVE ACTIVITY OF ACTIVE COMPONENTS OF CITRUS OILS Extensive research during the last half century has identified various molecular targets that can potentially be used not only for the prevention but also for treatment of cancer However, lack of success with targeted monotherapy resulting from bypass mechanisms has forced researchers to employ either combination therapy or agents that interfere with multiple cell-signaling pathways Increasing attention is paid to the possibility of applying cancer chemopreventive agents for individuals at high risk of neoplastic development (Aggarwal and Shishodia, 2006; Grassmann, 2005; Nishino et al., 2000; Tsuda et al., 2004; Wagner and Elmadfa, 2003) Natural compounds have practical advantages for this purpose with regard to availability, suitability for oral application, regulatory approval and mechanisms of action Recent studies have indicated that mechanisms underlying chemopreventive potential may be combinations of antioxidant, anti-inflammatory, immune-enhancing, and antihormone effects, with modification of drug-metabolizing enzymes, influence on the cell cycle and cell differentiation, induction of apoptosis and suppression of proliferation and angiogenesis playing roles in the initiation and secondary modification stages of neoplastic development (Figure 13.2) Accordingly, natural agents are advantageous for application to humans because of their combined action mechanisms (Wagner and Elmafda, 2003) However, while fruits and vegetables are recommended for prevention of cancer, their active ingredients (at the molecular level) and their mechanisms of action remain less well understood The monocyclic monoterpene limonene is a major constituent in several citrus oils (orange, lemon, mandarin, lime, and grapefruit) Because of its pleasant citrus fragrance, limonene is widely used as a fragrance and flavor additive in perfumes, soaps, foods, chewing gum, and beverages Plants biocompounds Normal cells Ini tiat ion Enhanced free radical generation Activation of phase I metabolizing enzymes Inactivation of phase II metabolizing enzyme Plants biocompounds Initiated cells n tio o om Pr Oncogenes Cell cycle alterations Overexpression of growth factors Tumor Growth Pr og re s sio n COX-2 Cytokines/TNF Angiogenic factors Adhesion molecules Matrix Metalloproteinases Plants biocompounds Tumor Metastasis FIGURE 13.2 Effects of plants biocompounds on the multistage carcinogenesis process K10034_C013.indd 539 9/21/2010 5:53:26 PM 540 Citrus Oils The Code of Federal Regulation lists d-limonene as generally recognized as safe (GRAS) for a flavoring agent Dietary intake of d-limonene varies depending on the types of foods consumed Hakim et al (2002) designed a study, by self-administration of a citrus food–frequency questionnaire, to assess the d-limonene content of different citrus juices and beverages and to develop a dietary-assessment instrument to measure d-limonene intake Mean intakes of d-limonene from citrus juices among consumers ranged between 13.0 and 13.2 mg/day d-Limonene has well-established chemopreventive activity against many types of cancers (Sun, 2007) and has demonstrated efficacy in preclinical models of breast and colon cancers Hakim et al (2000) carried out a case-control study to determine the usual citrus consumption patterns of an older Southwestern population and to evaluate how this citrus consumption varied with history of squamous cell carcinoma (SCC) of the skin In this Arizona population, citrus-peel consumption was not uncommon, with 34.7% of all subjects reporting citrus-peel use The authors found no association between the overall consumption of citrus fruits or juices and skin SCC, but the most striking feature was the protection purported by citrus-peel consumption, with a dose-response relationship Concerning skin cancer, it is interesting to note that limonene and perillic acid were proven to inhibit the metastatic progression of B16F-10 melanoma cells in C57BL/6 mice (Raphael and Kuttan, 2003a) Roberto et al (2010) evidenced that limonene could protect normal lymphocytes from diseases related to oxidative stress, including cancer In fact, limonene exerts antiproliferative action on a lymphoma cell line without modifying normal lymphocyte viability; in addition, it elicits a biphasic effect on proliferation of normal murine lymphocytes by decreasing H2O2 levels and increasing catalase and peroxidase activities, and protects murine lymphocytes against oxidative stress induced by exposure to H2O2 Limonene has been reported to induce apoptosis on tumor cells Hata et al (2003) reported that essential oils of sweet orange, grapefruit, and lemon, induce apoptosis in HL-60 cells because of limonene contained in them However, other components present in the essential oils of sweet orange and grapefruit may also be responsible for the observed apoptotic activity; in fact, the aldehyde compounds decanal, octanal, and citral present in the dichloromethane fraction proved to possess strong apoptotic activity Furthermore, d-limonene has antiangiogenic and proapoptotic effects on gastric cancer, thereby inhibits tumor growth and metastasis, as shown in a metastatic model simulating human gastric cancer and established by orthotopic implantation of histologically intact human tumor tissue into gastric wall of nude mice (Lu et al., 2004) Other mechanisms may be involved in the chemopreventive effect of limonene For example, Parija and Das (2003) demonstrated the involvement of the YY1 (Yin Yang 1) transcription factor in N-nitrodiethylamine-induced hepatocarcinogenesis, also showing that d-limonene mediated chemoprevention in this hepatocarcinogenesis model might be regulated by c-myc oncoprotein Consistent with these data, the chemopreventive effect of orally administrated orange oil (which contains 90% to 95% of limonene) was demonstrated on N-nitrosodiethylamine-induced hepatic preneoplasia in rats, together with restoration of the normal liver phenotype and upregulation of junctional complexes (Bodake et al., 2002) Limonene inhibits also hepatocarcinogenesis induced by N-nitrosomorpholine in male rats (Kaji et al., 2001); this effect may be clearly related to its effect in inhibiting cell proliferation and in enhancing apoptosis, but not through ras oncoprotein plasma membrane association d-Limonene induces phase I and phase II carcinogen-metabolizing enzymes (cytochrome p450), which metabolize carcinogens to less toxic forms and prevent the interaction of chemical carcinogens with DNA It has been shown to enhance gastrointestinal UDP-glucuronosyltransferase activity in rats (Van der Logt et al., 2004) Combination of d-limonene with cytotoxic agents or with other therapeutic approaches may be more effective than the employment of the monoterpene alone For example, d-limonene could improve the treatment outcome of hormone-refractory prostate cancer with docetaxel (Rabi and Bishayee, 2009), without being toxic to normal prostate epithelial cells and through the modulation K10034_C013.indd 540 9/21/2010 5:53:27 PM Biological Activities of Citrus Essential Oils 541 of proteins involved in mitochondrial pathway of apoptosis Furthermore the downregulation of the COL8A1 (collagen type VIII, α-1 gene) expression in the mouse hepatocarcinoma cell line Hca-F (which has a highly metastatic potential in the lymph nodes) also sensitized cells to the action of d-limonene (Zhao et al., 2009) Furthermore d-limonene modulates the immune response with significant potential for clinical application In fact, d-limonene increased the survival of lymphoma-bearing mice, and delayed hypersensitivity reaction to 2,4-dinitrofluorobenzene; furthermore it increased nitric oxide (NO) production in peritoneal macrophages obtained from these animals (Del Toro-Arreola et al., 2005) Furthermore, limonene demonstrated antiproliferative action also on a lymphoma cell line (BW5147), exerting a decrease in cell viability, that was related to apoptosis induction and increase in NO levels at long incubation times, and to cell arrest in different phases of the cell cycle at short times (Manuele et al., 2009) Finally, administration of various monoterpenes, including limonene, in Balb/c mice significantly increases the total antibody production, antibody producing cells in spleen, bone marrow cellularity and α-esterase positive cells, when compared to normal animals (Raphael and Kuttan, 2003b) The chemopreventive effect of d-limonene is not accompanied by significant toxicity; in particular, it is not genotoxic For example, limonene failed to increase the mutant frequency in the liver or kidney of male Big Blue rats exposed for 10 consecutive days to this monoterpene (Turner et al., 2001) Although male rats experienced an increased incidence of tubular cell hyperplasia, adenomas, and adenocarcinomas of the kidney, no evidence of carcinogenic activity was observed in female rats or male or female mice (National Toxicology Program, 2007) Furthermore, no induction of chromosomal aberrations or sister chromatid exchange in cultured Chinese hamster ovary cells was observed (National Toxicology Program, 2007) Finally, d-limonene elicited no mutagenicity in four strains of S typhimurium (TA98, TA100, TA1535, or TA1537) Similarly, the antigenotoxic effects of essential oil from Citrus aurantium L peels in combination with mutagenic metals and alkylating agents were evidenced by Demir et al (2009) using the wing spot test of Drosophila melanogaster exposed to potassium dichromate, cobalt chloride, ethylmethanesulfonate, and N-ethyl-N-nitrosourea as reference mutagens; the essential oil alone was not mutagenic and did not enhance the mutagenic effect of the reference mutagens, having also antigenotoxic effects in chronic cotreatments with all four mutagens Besides limonene, also other compounds contained in citrus essential oils, such as flavonoids and coumarins, are known for their chemopreventive properties Flavonoids are a ubiquitous family of phytochemicals that display a variety of biological effects, both beneficial and adverse depending on the individual compound Polymethoxylated flavones (PMFs) from citrus inhibit production of TNF-α and other pro-inflammatory cytokines As TNF-α also modulates Natural Killer (NK) cell activity, a mixture of PMFs purified from orange peel oil (consisting of nobiletin, tangeretin, trimethylscutellarein, sinensetin, 5-demethyl-nobiletin, hexa-O-methylquercetagetin, 5-demethyl-tetramethylscutellarein, and 5-hydroxy-3,3′,4′,6,7,8hexamethoxyflavone) was evaluated by Delaney et al (2001) to assess its potential to suppress, if orally administered, humoral, and innate immune functions in female B(6)C(3)F(1) mice sensitized to sheep red blood cells Long-term, high-dose exposure to this citrus PMF mixture caused a mild suppression of NK cell activity; however, humoral immunity was not sensitive to suppression at the same exposure levels Certain flavonoids are genotoxic, while others inhibit the genotoxicity of mutagenic agents However Delaney et al (2002) excluded the mutagenicity of the citrus PMF mixture described before by means of five bacterial tester strains (Salmonella typhimurium TA98, TA100, TA102, TA1535, and TA1537) either in the absence or presence of S9 activation Finally, the citrus coumarins isopimpinellin and imperatorin have chemopreventive effects when orally administered in SENCAR mice on skin-tumor initiation following topical application of benzo[a] pyrene and 7,12-dimethylbenz[a]anthracene, by blocking DNA adduct formation (Kleiner et al., 2002) K10034_C013.indd 541 9/21/2010 5:53:27 PM 542 13.7 Citrus Oils CITRUS ESSENTIAL OILS AND SKIN Citrus essential oils and/or their isolated components are efficiently used in dermatology As also reviewed in the second section of this chapter, one of their main applications concerns prevention and treatment of cutaneous infective diseases, due to the well-established antimicrobial properties of essential oils In addition, PUVA therapy continues to be the treatment of choice for patients with vitiligo, psoriasis, and other inflammatory skin diseases (Morison, 2004) As to the employment of citrus monoterpenes, such as limonene, in cosmetic and pharmaceutical industry, there is today increasing evidence that these compounds are suitable enhancer(s) for improving transdermal permeation of poorly absorbed drugs and may be useful for designing and discovering innovative transdermal drug systems In fact, chemicals offer tremendous potential in overcoming the skin barrier to enhance transport of drug molecules Individual chemicals are, however, limited in their efficacy in disrupting the skin barrier at low concentrations and usually cause skin irritation at high concentrations Multicomponent synergistic mixtures of chemicals (such as solvent mixtures, microemulsions, eutectic mixtures, complex self-assembled vesicles, inclusion complexes) have been shown to provide high skin permeabilization potency as compared to individual chemicals (Karande and Mitragotri, 2009) However, limonene, that is well known for its strong potential to act as skin penetration enhancer, and its oxidation products are also recognized as able to elicit allergic and irritative contact dermatitis 13.7.1 LIMONENE AS ENHANCER OF PERCUTANEOUS ABSORPTION Using skin as a port for systemic drug administration, transdermal drug delivery has expanded greatly over the last two decades The main advantage of this route is that it avoids the hepatic first-pass effect It is also recommended for some drugs in order to avoid problematic side effects However, transcutaneous delivery is heavily limited by the permeation characteristics of the stratum corneum, and for many drugs it is insufficient to obtain and maintain efficacious systemic levels Penetration enhancers are also a classical means for improving transdermal drug delivery when incorporated in transdermal therapeutic systems Much interest is at the moment focused on the use, as penetration enhancers, of molecules of natural origin, such as d-limonene and, in general, terpenes For example, d-limonene proved as good chemical enhancer by increasing the skin permeability of 6-mercaptopurine (Chandrashekar and Hiremath, 2008), and hydroxypropyl cellulose gel drug reservoir systems containing limonene act as optimal formulations for use in the design of membrane-controlled transdermal therapeutic system of ondansetron hydrochloride (Krishnaiah et al., 2008) Limonene may be efficiently used as penetration enhancer to improve skin permeation of carvedilol (Gannu et al., 2008a); furthermore, 8% v/w of d-limonene as a penetration enhancer and 20% v/w of dibutylphthalate as a plasticizer were used for the preparation of new, efficient, monolithic matrix-type transdermal drug-delivery systems for carvedilol, prepared using a film casting technique involving hydroxypropyl methylcellulose, hydroxypropyl cellulose, Eudragit RS 100, and Eudragit RL 100 as matrix-forming polymers (Gannu et al., 2008b) Güngör et al (2008) developed a matrix-type transdermal patches of verapamil hydrochloride with pectin as a matrix polymer, where nerolidol and limonene appeared to be the most promising enhancers It is well known that enhancers permeate into the skin and reversibly decrease the barrier resistance through different mechanisms The permeation enhancement of d-limonene and l-limonene to ligustrazine hydrochloride is due to multiple (very likely stereoselective) mechanisms including disordering and extracting the stratum corneum lipids (Zhang et al., 2006) Limonene is useful for enhancing the skin permeability of nimodipine and nicardipine from transdermal therapeutic systems containing hydroxypropyl methylcellulose gel or hydroxypropyl cellulose gel as reservoir respectively (Krishnaiah et al., 2002, 2004a,b), by inducing a partial extraction of lipids in the stratum corneum K10034_C013.indd 542 9/21/2010 5:53:27 PM Biological Activities of Citrus Essential Oils 543 Besides limonene, that can deliver haloperidol at a sustained percutaneous rate if incorporated in an organogel comprised of gelator GP1 and propylene glycol (Lim et al., 2006), also the oxide monoterpene limonene oxide can enhance the in vitro permeation of haloperidol through the human epidermis (Vaddi et al., 2003) The mode of interactions of this terpene with stratum corneum is different depending on the employed solvent systems In fact, limonene oxide in 50% v/v ethanol extracted stratum corneum lipids, disrupted the bilayer packing and partially fluidized the lipids, while, if dissolved in 100% v/v propylene glycol, disrupted the lipid bilayer and leaved the overall bilayer structure intact Besides to be employed as penetration enhancers for systemic drug administration, essential oils may be efficiently used for treating skin diseases by topical application In this case, percutaneous absorption of essential oils and oil components is of great interest So, the question is raised whether all components of a complex composed essential oil are equivalent with respect to their human skin permeation Schmitt et al (2009) investigated the cooperative effect of monoterpenes and phenylpropanoids on in vitro permeation through heat-separated human skin epidermis, clearly showing that cooperative effects of single essential oil components may influence percutaneous essential oil absorption In particular limonene showed an enhancing effect on the permeation of citronellol and eugenol 13.7.2 LIMONENE AND CONTACT DERMATITIS Limonene is one of the most commonly used fragrance compounds in western countries today When exposed to air, it autoxidizes, forming hydroperoxides that are strong contact allergens Several papers support to the European classification of R-(+)-limonene, containing oxidation products, as a skin sensitizer (Matura et al., 2002, 2003) However not only oxidized R-(+)- but also S-(–)-limonene is a common cause of allergic dermatitis contact (Matura et al., 2006) Furthermore, autoxidation of linalol and R-limonene appeared to have also an irritative effect, being oxidized linalol less irritating than oxidized R-limonene (Bråred Christensson et al., 2009) To cause allergic contact dermatitis, the hydroperoxides are considered to bind covalently to proteins in the skin via a radical mechanism When the radical formation from, and sensitizing potential of, allylic hydroperoxides found in the oxidation mixture of limonene, were examined, Johansson et al (2008) demonstrated that both carbon- and oxygen-centered radicals are important intermediates in the formation of hapten-protein complexes and that the sensitizing potency of the hydroperoxides is related to their structures Epidermal treatment with antioxidants, such as ascorbic acid or α-tocopherol, may have a protective effect on the allergenic effect of haptens that form full antigens via a radical mechanism, including limonene-2-hydroperoxide (Gäfvert et al., 2002) REFERENCES Aggarwal, B.B., and Shishodia, S 2006 Molecular targets of dietary agents for prevention and therapy of cancer Biochem Pharmacol 71(10):1397–421 Aloisi, A.M., Ceccarelli, I., Masi, F., and Scaramuzzino, A 2002 Effects of the essential oil from citrus lemon in male and female rats exposed to a persistent painful stimulation Behav Brain Res 136(1):127–35 Amantea, D., Fratto, V., Maida, S., et al 2009 Prevention of glutamate accumulation and upregulation of phospho-akt may account for neuroprotection afforded by bergamot essential oil against brain injury induced by focal cerebral ischemia in rat Int Rev Neurobiol 85:389–405 Anagnostopoulou, M.A., Kefalas, P., Papageorgiou, V.P., Assimopoulou, A.N., and Boskou, D 2006 Radical scavenging activity of various extracts and fractions of sweet orange peel (Citrus sinensis) Food Chem 94:19–25 Battey, A.S., Duffy, S., and Schaffner, D.W 2002 Modeling yeast spoilage in cold-filled ready-to-drink beverages with Saccharomyces cerevisiae, Zygosaccharomyces bailii, and Candida lipolytica Appl Environ Microbiol 68:1901–1906 K10034_C013.indd 543 9/21/2010 5:53:27 PM 544 Citrus Oils Battinelli, L., Mengoni, F., Lichtner, M., et al 2003 Effect of limonin and nomilin on HIV-1 replication on infected human mononuclear cells Planta Med 69:910–13 Belletti, N., Ndagijimana, M., Sisto, C., et al 2004 Evaluation of the antimicrobial activity of citrus essences on Saccharomyces cerevisiae J Agric Food Chem 52:6932–38 Belletti, N., Kamdem, S.S., Patrignani, F., et al 2007 Antimicrobial activity of aroma compounds against Saccharomyces cerevisiae and improvement of microbiological stability of soft drinks as assessed by logistic regression Appl Environ Microbiol 73:5580–6 Belletti, N., Lanciotti, R., Patrignani, F., and Gardini, F 2008 Antimicrobial efficacy of citron essential oil on spoilage and pathogenic microorganisms in fruit-based salads J Food Sci 73:M331–8 Berliocchi, L., Russo, R., Levato, A et al 2009 (–)-Linalol attenuates allodynia in neuropathic pain induced by spinal nerve ligation in c57/bl6 mice Int Rev Neurobiol 85:221–35 Bisignano, G., and Saija A 2002 The biological activity of citrus oils In Citrus eds G Dugo, and A Di Giacomo, 602–630 London and New York: Taylor & Francis Bodake, H.B., Panicker, K.N., Kailaje, V.V., and Rao, K.V 2002 Chemopreventive effect of orange oil on the development of hepatic preneoplastic lesions induced by N-nitrosodiethylamine in rats: an ultrastructural study Indian J Exp Biol 40(3):245–51 Bråred Christensson, J., Forsström, P., Wennberg, A.M., Karlberg, A.T., and Matura, M 2009 Air oxidation increases skin irritation from fragrance terpenes Contact Dermatitis 60(1):32–40 Burt, S 2004 Essential oils: their antibacterial properties and potential applications in foods—a review Int J Food Microbiol 94:223–253 Carvalho-Freitas, M.I.R., and Costa, M 2002 Anxiolytic and sedative effects of extracts and essential oil from Citrus aurantium L Biol Pharm Bull 25:1629–33 Ceccarelli, I., Masi, F., Fiorenzani, P., and Aloisi, A.M 2002 Sex differences in the citrus lemon essential oilinduced increase of hippocampal acetylcholine release in rats exposed to a persistent painful stimulation Neurosci Lett 330(1):25–8 Ceccarelli, I., Lariviere, W.R., Fiorenzani, P., Sacerdote, P., and Aloisi, A.M 2004 Effects of long-term exposure of lemon essential oil odor on behavioral, hormonal and neuronal parameters in male and female rats Brain Res 1001(1–2):78–86 Chandrashekar, N.S., and Hiremath, S.R 2008 In vivo immunomodulatory, cumulative skin irritation, sensitization and effect of d-limonene on permeation of 6-mercaptopurine through transdermal drug delivery Biol Pharm Bull 31(4):656–61 Choi, H.-S., Song, H.S., Ukeda, H., and Sawamura, M 2000 Radical scavenging activities of Citrus essential oils and their components: detection using 1,1-diphenyl-2-picrylhydrazyl J Agric Food Chem 48:4156–61 Choi, H.S 2006 Lipolytic effects of citrus peel oils and their components J Agric Food Chem 54(9):3254–8 Corasaniti, M.T., Maiuolo, J., Maida, S., et al 2007 Cell signaling pathways in the mechanisms of neuroprotection afforded by bergamot essential oil against NMDA-induced cell death in vitro Br J Pharmacol 151(4):518–29 Cosentino, S., Tuberoso, C.I.G., Pisano, B., et al 1999 In-vitro antimicrobial activity and chemical composition of Sardinian Thymus essential oils Lett Appl Microbiol 29:130–35 Cox, S.D., Mann, C.M., Markham, J.L., et al 2000 The mode of antimicrobial action of the essential oil of Melaleuca alternifolia (tea tree oil) J Appl Microbiol 88:170–5 Del Toro-Arreola, S., Flores-Torales, E., Torres-Lozano, C., et al 2005 Effect of d-limonene on immune response in BALB/c mice with lymphoma Int Immunopharmacol 5(5):829–38 Delaney, B., Phillips, K., Buswell, D., et al 2001 Immunotoxicity of a standardized citrus polymethoxylated flavone extract Food Chem Toxicol 39(11):1087–94 Delaney, B., Phillips, K., Vasquez, C., et al 2002 Genetic toxicity of a standardized mixture of citrus polymethoxylated flavones Food Chem Toxicol 40(5):617–24 Demir, E., Kocaogˇlu, S., Cetin, H., and Kaya, B 2009 Antigenotoxic effects of Citrus aurantium L fruit peel oil on mutagenicity of two alkylating agents and two metals in the Drosophila wing spot test Environ Mol Mutagen 50(6):483–8 Djordjevic, D., Cercaci, L., Alamed, J., McClements, D.J., and Decker, E.A 2007 Chemical and physical stability of citral and limonene in sodium dodecyl sulfate-chitosan and gum arabic-stabilized oil-in-water emulsions J Agric Food Chem 55(9):3585–91 Djordjevic, D., Cercaci, L., Alamed, J., McClements, D.J., and Decker, E.A 2008 Chemical and physical stability of protein and gum arabic-stabilized oil-in-water emulsions containing limonene J Food Sci 73(3):C167–72 Dorman, H.J.D., and Deans, S.G 2000 Antimicrobial agents from plant: antibacterial activity of plant volatile oils J Appl Microbiol 88(2):308–16 K10034_C013.indd 544 9/21/2010 5:53:27 PM Biological Activities of Citrus Essential Oils 545 Dusan, F., Marian, S., Katarina, D., and Dobroslava, B 2006 Essential oils e their antimicrobial activity against Escherichia coli and effect on intestinal cell viability Toxicol in Vitro 20(8):1435–45 Edris, A.E 2007 Pharmaceutical and therapeutic potentials of essential oils and their individual volatile constituents: a review Phytother Res 21(4):308–23 Fisher, K., and Phillips, C 2006 The effect of lemon, orange and bergamot essential oils and their components on the survival of Campylobacter jejuni, Escherichia coli O157, Listeria monocytogenes, Bacillus cereus and Staphylococcus aureus in vitro and in food systems J Appl Microbiol 101(6):1232–40 Fisher, K., and Phillips, C 2009 The mechanism of action of a citrus oil blend against Enterococcus faecium and Enterococcus faecalis J Appl Microbiol 106(4):1343–9 Fukumoto, S., Sawasaki, E., Okuyama, S., Miyake, Y., and Yokogoshi, H 2006 Flavor components of monoterpenes in citrus essential oils enhance the release of monoamines from rat brain slices Nutr Neurosci 9(1–2):73–80 Gäfvert, E., Nilsson, J.L., Hagelthorn, G., and Karlberg, A.T 2002 Free radicals in antigen formation: reduction of contact allergic response to hydroperoxides by epidermal treatment with antioxidants Br J Dermatol 146(4):649–56 Gannu, R., Vishnu, Y.V., Kishan, V., and Rao, Y.M 2008a In vitro permeation of carvedilol through porcine skin: effect of vehicles and penetration enhancers PDA J Pharm Sci Technol 62(4):256–63 Gannu, R., Vishnu, Y.V., Kishan, V., and Rao, Y.M 2008b Development of carvedilol transdermal patches: evaluation of physicochemical, ex vivo and mechanical properties PDA J Pharm Sci Technol 62(6):391–401 Gardini, F., Lanciotti, R., Sinigaglia, M., and Guerzoni, M.E 1997 A headspace gas chromatographic approach for the monitoring of the microbial cell activity and the cell viability evaluation J Microbiol Methods 29:103–14 Girennavar B., Jayaprakasha G.K., Jadegoud Y., Nagana Gowda G.A., and Patil B.S 2007 Radical scavenging and cytochrome P450 3A4 inhibitory activity of bergaptol and geranylcoumarin from grapefruit Bioorg Med Chem 15(11):3684–91 Grassmann, J., Schneider, D., Weiser, D., and Elstner, E.F 2001 Antioxidative effects of lemon oil and its components on copper induced oxidation of low density lipoprotein Arzneimittelforschung 51(10):799–805 Grassmann, J 2005 Terpenoids as plant antioxidants Vitam Horm 72:505–35 Güngör, S., Bektas¸, A., Alp, F.I et al 2008 Matrix-type transdermal patches of verapamil hydrochloride: in vitro permeation studies through excised rat skin and pharmacodynamic evaluation in rats Pharm Dev Technol 13(4):283–9 Hakim, I.A., Harris, R.B., and Ritenbaugh, C 2000 Citrus peel use is associated with reduced risk of squamous cell carcinoma of the skin Nutr Cancer 37:161–168 Hakim, I.A., Hartz, V., Graver, E., Whitacre, R., and Alberts, D 2002 Development of a questionnaire and a database for assessing dietary d-limonene intake Public Health Nutr 5(6A):939–45 Hata, T., Sakaguchi, I., Mori, M et al 2003 Induction of apoptosis by Citrus paradisi essential oil in human leukemic (HL-60) cells In Vivo 17(6):553–9 Haze, S., Sakai, K., and Gozu, Y 2002 Effects of fragrance inhalation on sympathetic activity in normal adults Jpn J Pharmacol 90(3):247–53 Holley, R.A., and Patel, D 2005 Improvement in shelf-life and safety of perishable foods by plant essential oils and smoke antimicrobials Food Microbiol 22(4):273–92 Holm, L., and Fitzmaurice, L 2008 Emergency department waiting room stress: can music or aromatherapy improve anxiety scores? Pediatr Emerg Care 24(12):836–8 Johansson, S., Giménez-Arnau, E., Grøtli, M., Karlberg, A.T., and Börje, A 2008 Carbon- and oxygencentered radicals are equally important haptens of allylic hydroperoxides in allergic contact dermatitis Chem Res Toxicol 21(8):1536–47 Kaji, I., Tatsuta, M., Iishi, H., Baba, M., Inoue, A., and Kasugai, H 2001 Inhibition by d-limonene of experimental hepatocarcinogenesis in Sprague-Dawley rats does not involve p21(ras) plasma membrane association Int J Cancer 93(3):441–4 Karande, P., and Mitragotri, S 2009 Enhancement of transdermal drug delivery via synergistic action of chemicals Biochim Biophys Acta 1788(11):2362–73 Kelly, C., Jewell, C., and O’Brien, N.M 2003 The effect of dietary supplementation with the citrus limonoids, limonin and nomilin on xenobiotic-metabolizing enzymes in the liver and small intestine of the rat Nutr Res 23:681–90 Kleiner, H.E., Vulimiri, S.V., Starost, M.F., Reed, M.J., and Di Giovanni, J., 2002 Oral administration of the citrus coumarin, isopimpinellin, blocks DNA adduct formation and skin tumor initiation by 7,12dimethylbenz[a]anthracene in SENCAR mice Carcinogenesis 23(10):1667–75 K10034_C013.indd 545 9/21/2010 5:53:27 PM 546 Citrus Oils Komiya, M., Takeuchi, T., and Harada, E 2006 Lemon oil vapor causes an anti-stress effect via modulating the 5-HT and DA activities in mice Behav Brain Res 172(2):240–9 Koo, H.N., Hong, S.H., Kim, C.Y., et al 2002 Inhibitory effect of apoptosis in human astrocytes CCF-STTG1 cells by lemon oil Pharmacol Res 45(6):469–73 Krishnaiah, Y.S., Satyanarayana, V., and Bhaskar, P 2002 Effect of limonene on the in vitro permeation of nicardipine hydrochloride across the excised rat abdominal skin Pharmazie 57(12): 842–7 Krishnaiah, Y.S., Bhaskar, P., and Satyanarayana, V 2004a In vitro percutaneous permeability enhancement of nimodipine by limonene across the excised rat abdominal skin Pharmazie 59(12):942–7 Krishnaiah, Y.S., Bhaskar, P., and Satyanarayana, V 2004b Formulation and evaluation of limonene-based membrane-moderated transdermal therapeutic system of nimodipine Drug Deliv 11(1):1–9 Krishnaiah, Y.S., Raju, V., Shiva Kumar, M., et al 2008 Studies on optimizing in vitro transdermal permeation of ondansetron hydrochloride using nerodilol, carvone, and limonene as penetration enhancers Pharm Dev Technol 13(3):177–85 Lehrner, J., Eckersberger, C., Walla, P., Pötsch, G., and Deecke, L 2000 Ambient odor of orange in a dental office reduces anxiety and improves mood in female patients Physiol Behav 71(1–2):83–6 Lim, P.F., Liu, X.Y., Kang, L., Ho, P.C., Chan, Y.W., and Chan, S.Y 2006 Limonene GP1/PG organogel as a vehicle in transdermal delivery of haloperidol Int J Pharm 311(1–2):157–64 Louriero, V., and Querol, A 1999 The prevalence and control of spoilage yeasts in foods and beverages Trends Food Sci Technol 10:356–65 Lu, X.G., Zhan, L.B., Feng, B.A., Qu, M.Y., Yu, L.H., and Xie, J.H 2004 Inhibition of growth and metastasis of human gastric cancer implanted in nude mice by d-limonene World J Gastroenterol 10(14):2140–4 Malhotra, S., Suri, S., and Tuli, R 2009 Antioxidant activity of citrus cultivars and chemical composition of Citrus karma essential oil Planta Med 75(1):62–4 Manners, G.D 2007 Citrus limonoids: analysis, bioactivity, and biomedical prospects J Agric Food Chem 55(21):8285–94 Manthey, J.A., and Bendele, P 2008 Anti-inflammatory activity of an orange peel polymethoxylated flavone, 3’,4’,3,5,6,7,8-heptamethoxyflavone, in the rat carrageenan/paw edema and mouse lipopolysaccharidechallenge assays J Agric Food Chem 56(20):9399–403 Manuele, M.G., Arcos, M.L., Davicino, R., Ferraro, G., Cremaschi, G., and Anesini, C 2009 Mechanism of the antiproliferative action of limonene on a lymphoma cell line: participation of nitric oxide Phytother Res 23(7):1011–7 Matura, M., Goossens, A., Bordalo, O., et al 2002 Oxidized citrus oil (R-limonene): a frequent skin sensitizer in Europe J Am Acad Dermatol 47(5):709–14 Matura, M., Goossens, A., Bordalo, O., et al 2003 Patch testing with oxidized R-(+)-limonene and its hydroperoxide fraction Contact Dermatitis 49(1):15–21 Matura, M., Sköld, M., Börje, A., et al 2006 Not only oxidized R-(+)- but also S-(–)-limonene is a common cause of contact allergy in dermatitis patients in Europe Contact Dermatitis 55(5):274–9 Meadows, M 2006 September to October 2006, posting date Benzene in beverages FDA Consumer Misharina, T.A., and Samusenko, A.L 2008 Antioxidant properties of essential oils from lemon, grapefruit, coriander, clove, and their mixtures Prikl Biokhim Mikrobiol 44(4):482–6 Miyazawa, M., Tougo, H., and Ishihara, M 2001 Inhibition of acetylcholinesterase activity by essential oil from Citrus paradisi Nat Prod Lett 15(3):205–10 Mizrahi, B., Shapira, L., Domb, A.J., and Houri-Haddad, Y 2006 Citrus oil and MgCl2 as antibacterial and anti-inflammatory agents J Periodontol 77(6):963–8 Mollace, V., Ragusa, S., Sacco, I., et al 2008 The protective effect of bergamot oil extract on lecitine-like oxyLDL receptor-1 expression in balloon injury-related neointima formation J Cardiovasc Pharmacol Ther 13(2):120–9 Moraes, T.M., Kushima, H., Moleiro, F.C., et al 2009 Effects of limonene and essential oil from Citrus aurantium on gastric mucosa: role of prostaglandins and gastric mucus secretion Chem Biol Interact 180(3):499–505 Morison, W.L 2004 Psoralen ultraviolet A therapy in 2004 Photodermatol Photoimmunol Photomed 20(6):315–20 Morrone, L.A., Rombolà, L., Pelle, C., et al 2007 The essential oil of bergamot enhances the levels of amino acid neurotransmitters in the hippocampus of rat: implication of monoterpene hydrocarbons Pharmacol Res 55(4):255–62 National Toxicology Program 2007 Toxicology and Carcinogenesis Studies of d-Limonene (CAS No 5989– 27–5) in F344/N Rats and B6C3F1 Mice http://ntp.niehs.nih.gov/index.cfm?objectid=07086449–9787 –5414–556E052773467BE9 Last accessed April 4, 2010 K10034_C013.indd 546 9/21/2010 5:53:27 PM Biological Activities of Citrus Essential Oils 547 Niijima, A., and Nagai, K 2003 Effect of olfactory stimulation with flavor of grapefruit oil and lemon oil on the activity of sympathetic branch in the white adipose tissue of the epididymis Exp Biol Med 228(10):1190–2 Nishino, H., Tokuda, H., Satomi, Y., et al 2000 Cancer chemoprevention by phytochemicals and their related compounds Asian Pac J Cancer Prev 1(1):49–55 Parija, T., and Das, B.R 2003 Involvement of YY1 and its correlation with c-myc in NDEA induced hepatocarcinogenesis, its prevention by d-limonene Mol Biol Rep 30(1):41–6 Perez-Cacho, P.R., and Rouseff, R 2008 Processing and storage effects on orange juice aroma: a review J Agric Food Chem 56(21):9785–96 Pultrini Ade, M., Galindo, L.A., and Costa, M 2006 Effects of the essential oil from Citrus aurantium L in experimental anxiety models in mice Life Sci 78(15):1720–5 Rabi, T., and Bishayee, A 2009 d-Limonene sensitizes docetaxel-induced cytotoxicity in human prostate cancer cells: generation of reactive oxygen species and induction of apoptosis J Carcinog 8:9 Rangan, C., and Barceloux, D.G 2009 Food additives and sensitivities Dis Mon 55(5):292–311 Raphael, T.J., and Kuttan, G 2003a Effect of naturally occurring monoterpenes carvone, limonene and perillic acid in the inhibition of experimental lung metastasis induced by B16F-10 melanoma cells J Exp Clin Cancer Res 22(3):419–24 Raphael, T.J., and Kuttan, G 2003b Immunomodulatory activity of naturally occurring monoterpenes carvone, limonene, and perillic acid Immunopharmacol Immunotoxicol 25(2):285–94 Rios, J.L., and Recio, M.C 2005 Medicinal plants and antimicrobial activity J Ethnopharm 100:80–4 Roberto, D., Micucci, P., Sebastian, T., Graciela, F., and Anesini, C 2010 Antioxidant activity of limonene on normal murine lymphocytes: relation to HO modulation and cell proliferation Basic Clin Pharmacol Toxicol 106(1):38–44 Romano, L., Battaglia, F., Masucci, L., et al 2005 In vitro activity of bergamot natural essence and furocoumarin-free and distilled extracts, and their associations with boric acid, against clinical yeast isolates J Antimicrob Chemother 55:110–4 Rombolà, L., Corasaniti, M.T., Rotiroti, D., et al 2009 Effects of systemic administration of the essential oil of bergamot (BEO) on gross behavior and EEG power spectra recorded from the rat hippocampus and cerebral cortex Funct Neurol 24(2):107–12 Sakurada, T., Kuwahata, H., Katsuyama, S., et al 2009 Intraplantar injection of bergamot essential oil into the mouse hindpaw effects on capsaicin-induced nociceptive behaviors Int Rev Neurobiol 85:237–48 Sanguinetti, M., Posteraro, B., Romano, L., et al 2007 In vitro activity of Citrus bergamia (bergamot) oil against clinical isolates of dermatophytes J Antimicrob Chemother 59(2):305–8 Schmitt, S., Schaefer, U.F., Doebler, L., and Reichling, J 2009 Cooperative interaction of monoterpenes and phenylpropanoids on the in vitro human skin permeation of complex composed essential oils Planta Med 75(13):1381–5 Sharma, N., and Tripathi, A 2008 Effects of Citrus sinensis (L.) Osbeck epicarp essential oil on growth and morphogenesis of Aspergillus niger (L.) Van Tieghem Microbiol Res 163(3):337–44 Shen, J., Niijima, A., Tanida, M., Horii, Y., Maeda, K., and Nagai, K 2005 Olfactory stimulation with scent of grapefruit oil affects autonomic nerves, lipolysis and appetite in rats Neurosci Lett 380(3):289–94 Smith-Palmer, A., Stewart, J., and Fyfe, L 2001 The potential application of plant essential oils as natural food preservatives in soft cheese Food Microbiol 18(4):463–70 Suhr, K.I., and Nielsen, P.V 2003 Antifungal activity of essential oils evaluated by two different application techniques against rye bread spoilage fungi J Appl Microbiol 94:665–74 Sun, J 2007 d-Limonene: safety and clinical applications Altern Med Rev 12(3):259–64 Takahashi, Y., Inaba, N., Kuwahara, S., and Kuki, W 2003 Antioxidative effect of Citrus essential oil components on human low-density lipoprotein in vitro Biosci Biotechnol Biochem 67(1):195–7 Takikawa, A., Abe, K., Yamamoto, M., et al 2002 Antimicrobial activity of nutmeg against Escherichia coli O157 J Biosci Bioeng 94:315–20 Tanida, M., Niijima, A., Shen, J., Nakamura, T., and Nagai, K 2005 Olfactory stimulation with scent of essential oil of grapefruit affects autonomic neurotransmission and blood pressure Brain Res 1058(1–2):44–55 Tassou, C., Koutsoumanis, K., and Nychas, G.J.E 2000 Inhibition of Salmonella enteritidis and Staphylococcus aureus in nutrient broth by mint essential oil Food Res Intern 33:273–80 Tirkey, N., Pilkhwal, S., Kuhad, A., and Chopra, K 2005 Hesperidin, a citrus bioflavonoid, decreases the oxidative stress produced by carbon tetrachloride in rat liver and kidney BMC Pharmacol 5:2 K10034_C013.indd 547 9/21/2010 5:53:27 PM 548 Citrus Oils Tsuda, H., Ohshima, Y., Nomoto, H., et al 2004 Cancer prevention by natural compounds Drug Metab Pharmacokinet 19(4):245–63 Turner, S.D., Tinwell, H., Piegorsch, W., Schmezer, P., and Ashby, J 2001 The male rat carcinogens limonene and sodium saccharin are not mutagenic to male big blue rats Mutagenesis 16:329–332 Vaddi, H.K., Ho, P.C., Chan, Y.W., and Chan, S.Y 2003 Oxide terpenes as human skin penetration enhancers of haloperidol from ethanol and propylene glycol and their modes of action on stratum corneum Biol Pharm Bull 26(2):220–8 Van der Logt, E.M., Roelofs, H.M., van Lieshout, E.M., Nagengast, F.M., and Peters, W.H 2004 Effects of dietary anticarcinogens and nonsteroidal anti-inflammatory drugs on rat gastrointestinal UDPglucuronosyltransferases Anticancer Res 24:843–9 Wagner, K.H., and Elmadfa, I 2003 Biological relevance of terpenoids Overview focusing on mono-, di- and tetraterpenes Ann Nutr Metab 47(3–4):95–106 Wuryatmo, E., Klieber, A., and Scott E.S 2003 Inhibition of citrus postharvest pathogens by vapor of citral and related compounds in culture J Agric Food Chem 51:2637–40 Yip, Y.B., and Tam, A.C 2008 An experimental study on the effectiveness of massage with aromatic ginger and orange essential oil for moderate-to-severe knee pain among the elderly in Hong Kong Complement Ther Med 16(3):131–8 Zhang, C.F., Yang, Z.L., and Luo, J.B 2006 Effects of d-limonene and l-limonene on transdermal absorption of ligustrazine hydrochloride Yao Xue Xue Bao 41(8):772–7 Zhao, Y., Jia, L., Mao, X., Xu, H., Wang, B., and Liu, Y 2009 siRNA-targeted COL8A1 inhibits proliferation, reduces invasion and enhances sensitivity to d-limonene treatment in hepatocarcinoma cells IUBMB Life 61(1):74–9 Zhou, W., Fukumoto, S., and Yokogoshi, H 2009 Components of lemon essential oil attenuate dementia induced by scopolamine Nutr Neurosci 12(2):57–64 K10034_C013.indd 548 9/21/2010 5:53:27 PM .. .Citrus Oils Composition, Advanced Analytical Techniques, Contaminants, and Biological Activity K10034_C000.indd i 9/21/2010 8:32:24 PM Medicinal and Aromatic Plants — Industrial... 49 Citrus Oils, edited by Giovanni Dugo and Luigi Mondello 9/21/2010 8:32:26 PM K10034_C000.indd iv 9/21/2010 8:32:26 PM Citrus Oils Composition, Advanced Analytical Techniques, Contaminants, and. .. 978-1-4398-0028-7 Citrus oils Composition Citrus oils Analysis Citrus oils Therapeutic use I Dugo, Giovanni II Mondello, Luigi III Title: Composition, analysis, contamination and properties of citrus oils