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Natural Product Research: Formerly Natural Product Letters Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/gnpl20
Antioxidant and antifungal activity of different extracts obtained from aerial parts of Inula crithmoides L. a
b
c
Anahi Bucchini , Donata Ricci , Federica Messina , Maria Carla c
c
a
Marcotullio , Massimo Curini & Laura Giamperi a
Dipartimento di Scienze della Terra, della Vita e dell'Ambiente – Università di Urbino “Carlo Bo”, Via Bramante 28, 61029Urbino (PU), Italy b
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Dipartimento di Scienze Biomolecolari, Università di Urbino “Carlo Bo”, Via Bramante 28, 61029Urbino (PU), Italy c
Dipartimento di Scienze Farmaceutiche, Università di Perugia, Via del Liceo 1, 06123Perugia (PG), Italy Published online: 26 Nov 2014.
To cite this article: Anahi Bucchini, Donata Ricci, Federica Messina, Maria Carla Marcotullio, Massimo Curini & Laura Giamperi (2014): Antioxidant and antifungal activity of different extracts obtained from aerial parts of Inula crithmoides L., Natural Product Research: Formerly Natural Product Letters, DOI: 10.1080/14786419.2014.983102 To link to this article: http://dx.doi.org/10.1080/14786419.2014.983102
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content.
Downloaded by [Nova Southeastern University] at 08:25 06 January 2015
This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/termsand-conditions
Natural Product Research, 2014 http://dx.doi.org/10.1080/14786419.2014.983102
SHORT COMMUNICATION Antioxidant and antifungal activity of different extracts obtained from aerial parts of Inula crithmoides L. Anahi Bucchinia, Donata Riccib, Federica Messinac, Maria Carla Marcotullioc, Massimo Curinic and Laura Giamperia*
Downloaded by [Nova Southeastern University] at 08:25 06 January 2015
a
Dipartimento di Scienze della Terra, della Vita e dell’Ambiente – Universita` di Urbino “Carlo Bo”, Via Bramante 28, 61029 Urbino (PU), Italy; bDipartimento di Scienze Biomolecolari, Universita` di Urbino “Carlo Bo”, Via Bramante 28, 61029 Urbino (PU), Italy; cDipartimento di Scienze Farmaceutiche, Universita` di Perugia, Via del Liceo 1, 06123 Perugia (PG), Italy (Received 22 May 2014; final version received 26 October 2014)
The total phenolic content, antioxidant and antifungal activities of three Inula crithmoides extracts (n-hexane, methylene chloride and MeOH) were investigated. The methanolic extract showed the highest total phenolic content. In the DPPH assay, the methanolic and hexane extracts exhibited the highest DPPH-radical scavenging activity; in the 5-lipoxygenase assay, the hexane extract showed greater inhibitory effect with an IC50 similar to that of Trolox and ascorbic acid. The antifungal activity of the methanolic extract revealed a higher activity against Phytophtora cryptogea and Alternaria solani. Keywords: Inula crithmoides; phenolics; antioxidant activity; antifungal activity
1. Introduction Inula crithmoides L. is a plant from the halophyte group and belonging to the Compositae family. It is widespread in the Mediterranean basin, in maritime meadows and saltwater marshes, and in Italy it grows along the coasts. The genus Inula for its richness in secondary metabolites has been the subject of many recent researche’ studies, which highlighted the different bioactivities of several extracts obtained from this plant, due to different secondary metabolites content. Some Inula spp. contain interesting compounds with antibacterial, antifungal, cytotoxic and antioxidant activities (Mosaad et al. 2008). In a recent paper, *Corresponding author. Email: [email protected] This manuscript was submitted at SIF 2013, XIII National Congress of Societa` Italiana di Fitochimica (Italian Phytochemical Society), Gargnano (Italy), 19 –21 September 2013. q 2014 Taylor & Francis
Downloaded by [Nova Southeastern University] at 08:25 06 January 2015
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A. Bucchini et al.
Omezzine et al. (2011) showed a good antifungal activity of different Inula extracts against two Trichoderma species and three formae speciales of Fusarium oxysporum. Furthermore, Giamperi et al. (2010) showed a good antioxidant activity of the essential oil obtained from I. crithmoides aerial parts and a considerable antifungal activity of extracts obtained from callus cultures. Fungi and bacteria have unfavourable effects on quality, safety and preservation of food. Synthetic chemicals are widely used in the control of plant diseases; however, these compounds may leave toxic residues in treated products (Zhao et al. 2010). Therefore, there is an urgent demand for the discovery of new safer and more effective antimicrobial substances, and medicinal plants can be regarded as a valid and alternative source of bioactive compounds (Bucchini et al. 2013). Among these, the components with phenolic structures such as carvacrol, eugenol and thymol were highly active against pathogens and showed a good antioxidant activity. The aim of our work was to evaluate the total phenolic content, antioxidant and antifungal activities of three different extracts obtained from I. crithmoides aerial parts.
2. Results and discussion 2.1. Total phenolic content Powdered dried aerial parts were sequentially extracted by maceration with n-hexane (H), methylene chloride (DCM) and methanol (MeOH). Yields of extractions are reported in Table S1. Looking at the same table it is evident that among the three Inula extracts, the methanol extract showed the highest total phenolic content, 15.52 mg/g dry weight of starting material (DW). Furthermore, the total polyphenols content exhibited the descending order among MeOH . H . DCM extracts.
2.2. Free-radical scavenging activity The DPPH radical assay has been widely used to test the ability of compounds as free-radical scavengers or hydrogen donors and to evaluate the antioxidative activity of plant extracts and foods (Prior et al. 2005; Giamperi et al. 2012; Seow et al. 2012). Our results (Table S1) show that the methanol and hexane extracts exhibited the highest DPPH-radical scavenging activity with EC50 value of 0.59 and 0.57 mg DW/mL, respectively. These EC50 values are six times higher than that of the ascorbic acid (0.110 mg DW/mL). Antioxidant activity assessed with the DPPH test in both extracts is linearly related with polyphenols content (R 2 is 0.99 for methanol extract and is 0.97 for hexane extract).
2.3. Anti- inflammatory activity (inhibition of lipid peroxidation) 5-Lipoxygenase (5-LOX) is one of the key enzymes in the synthesis of leukotrienes, inflammatory eicosanoids and histamine from mast cells, and in the formation of cytokines (mainly TNF-a, IL-6) by macrophages followed by the liberation ROS (reactive oxygen species). Thus, a reduction of the production of inflammatory mediators by inhibiting the activities of 5-lipoxygenase or cyclooxygenase in addition to a direct ROS scavenging by antioxidants is considered as a promising therapeutic target (El-Readi et al. 2013). The aim of this step of our work was to evaluate the inhibitory effect of different I. crithmoides extracts on 5-lipoxygenase. The hexane extract showed greater inhibitory effect with an IC50 similar to that of Trolox and ascorbic acid (IC50 ¼ 13.48, 11.89 and 18.63 mg DW/ mL, respectively) (Table S1).
Downloaded by [Nova Southeastern University] at 08:25 06 January 2015
Natural Product Research
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2.4. Antifungal activity Recently, in different parts of the world, attention has been paid towards exploitation of higher plant products as novel chemotherapeutants in plant protection (Kotan et al. 2010). For this reason we decided to test the obtained extracts for their antifungal effect against eight phytopathogenic fungi and their possible activity in control of fungal diseases. The antifungal activity was tested in vitro by using the agar dilution method. Extracts demonstrated a good antifungal activity against Phytophtora cryptogea and Alternaria solani. Particularly, the antifungal activity of methanol extract at the dose of 400 ppm was comparable with the activity of the positive control (Nystatin 50 ppm). Also the hexane extract exhibited a good antifungal activity. In this case the dose of 400 ppm of the extract showed an important antifungal activity when compared with the untreated control with a similar trend of the methanol extract. A weaker activity was shown against all fungi of the genus Fusarium. Only for F. oxysporum, the fungicidal activity of extracts was similar to that shown towards A. solani (Tables S2 –S4). The DCM extract is not effective against all fungi tested (data not shown).
3. Conclusions The present report is the first comprehensive study of the polyphenolic content, antioxidant, anti-inflammatory and antifungal activities of I. crithmoides extracts. Interestingly, the antiinflammatory activity of I. critmoides methanol and hexane extracts positively correlated with their polyphenolic content (R 2 ¼ 0.94 and 0.95, respectively). This study indicated that the methanolic and the hexane extracts of I. crithmoides might be regarded as natural antioxidant and anti-inflammatory agents. In agriculture, these extracts might be applicable as natural chemicals for integrated plant protection against a variety of phytopathogenic fungi that cause an economically important tomato and potato disease. Further studies are required to investigate the in vivo efficacy. With efficient collaborations with pharmacologists and medical doctors, plant pathologists will be crucial to see the development of a comprehensive study.
Supplementary material Experimental details relating to this paper are available online, alongside Tables S1 – S4.
References Bucchini A, Giamperi L, Ricci D. 2013. Total polyphenol content, in vitro antifungal and antioxidant activities of callus cultures from Inula crithmoides L. Nat Prod Commun. 8:1587–1590. El-Readi MZ, Eid HH, Ashour ML, Eid SY, Labib RM, Sporer F, Wink M. 2013. Variations of the chemical composition and bioactivity of essential oils from leaves and stems of Liquidambar styraciflua (Altingiacae). J Pharm Pharmacol. 65:1653 –1663. Giamperi L, Bucchini A, Bisio A, Giacomelli E, Romussi G, Ricci D. 2012. Total phenolic content and antioxidant activity of Salvia spp. exudates. Nat Prod Commun. 7:201–202. Giamperi L, Bucchini A, Fraternale D, Genovese S, Curini M, Ricci D. 2010. Composition and antioxidant activity of Inula crithmoides essential oil grown in central Italy (Marche Region). Nat Prod Commun. 5:315–318. Kotan R, Cakir A, Dadasoglu F, Aydin T, Cakmakci R, Ozer H, Kordali S, Mete E, Dikbas N. 2010. Antibacterial activities of essential oils and extracts of Turkish Achillea, Satureja and Thymus species against plant pathogenic bacteria. J Sci Food Agric. 90:145–160. Mosaad A, Abdel-Wahhab S, Abdel-Azim H, Aziza AEN. 2008. Inula crithmoides extract protects against ochratoxin Ainduced oxidative stress, clastogenic and mutagenic alteration in male rats. Toxicon. 52:566–573. Omezzine F, Daami-Remadi M, Rinez A, Ladhari A, Haoulala R. 2011. In vitro assessment of Inula spp. organic extracts for their antifungal activity against some pathogenic and antagonistic fungi. Afr J Microbiol Res. 5:3527–3531.
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A. Bucchini et al.
Downloaded by [Nova Southeastern University] at 08:25 06 January 2015
Prior RL, Wu X, Schaich K. 2005. Standardized methods for the determination of antioxidant capacity and phenolic in foods and dietary supplements. J Agric Food Chem. 53:4290–4302. Seow MH, Amru NB, Chandran S. 2012. Antioxidant activity, phenolic and flavonoid contents in the leaves of different varieties of sweet potato (Ipomoea batatas). Aust J Crop Sci. 6:375–380. Zhao J, Li Y, Liu Q, Gao K. 2010. Antimicrobial activities of some thymol derivatives from the roots of Inula hupehensis. Food Chem. 120:512–516.
Natural Product Research: Formerly Natural Product Letters Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/gnpl20
Antioxidant and antifungal activity of different extracts obtained from aerial parts of Inula crithmoides L. a
b
c
Anahi Bucchini , Donata Ricci , Federica Messina , Maria Carla c
c
a
Marcotullio , Massimo Curini & Laura Giamperi a
Dipartimento di Scienze della Terra, della Vita e dell'Ambiente – Università di Urbino “Carlo Bo”, Via Bramante 28, 61029Urbino (PU), Italy b
Click for updates
Dipartimento di Scienze Biomolecolari, Università di Urbino “Carlo Bo”, Via Bramante 28, 61029Urbino (PU), Italy c
Dipartimento di Scienze Farmaceutiche, Università di Perugia, Via del Liceo 1, 06123Perugia (PG), Italy Published online: 26 Nov 2014.
To cite this article: Anahi Bucchini, Donata Ricci, Federica Messina, Maria Carla Marcotullio, Massimo Curini & Laura Giamperi (2014): Antioxidant and antifungal activity of different extracts obtained from aerial parts of Inula crithmoides L., Natural Product Research: Formerly Natural Product Letters, DOI: 10.1080/14786419.2014.983102 To link to this article: http://dx.doi.org/10.1080/14786419.2014.983102
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content.
Downloaded by [Nova Southeastern University] at 08:25 06 January 2015
This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/termsand-conditions
Natural Product Research, 2014 http://dx.doi.org/10.1080/14786419.2014.983102
SHORT COMMUNICATION Antioxidant and antifungal activity of different extracts obtained from aerial parts of Inula crithmoides L. Anahi Bucchinia, Donata Riccib, Federica Messinac, Maria Carla Marcotullioc, Massimo Curinic and Laura Giamperia*
Downloaded by [Nova Southeastern University] at 08:25 06 January 2015
a
Dipartimento di Scienze della Terra, della Vita e dell’Ambiente – Universita` di Urbino “Carlo Bo”, Via Bramante 28, 61029 Urbino (PU), Italy; bDipartimento di Scienze Biomolecolari, Universita` di Urbino “Carlo Bo”, Via Bramante 28, 61029 Urbino (PU), Italy; cDipartimento di Scienze Farmaceutiche, Universita` di Perugia, Via del Liceo 1, 06123 Perugia (PG), Italy (Received 22 May 2014; final version received 26 October 2014)
The total phenolic content, antioxidant and antifungal activities of three Inula crithmoides extracts (n-hexane, methylene chloride and MeOH) were investigated. The methanolic extract showed the highest total phenolic content. In the DPPH assay, the methanolic and hexane extracts exhibited the highest DPPH-radical scavenging activity; in the 5-lipoxygenase assay, the hexane extract showed greater inhibitory effect with an IC50 similar to that of Trolox and ascorbic acid. The antifungal activity of the methanolic extract revealed a higher activity against Phytophtora cryptogea and Alternaria solani. Keywords: Inula crithmoides; phenolics; antioxidant activity; antifungal activity
1. Introduction Inula crithmoides L. is a plant from the halophyte group and belonging to the Compositae family. It is widespread in the Mediterranean basin, in maritime meadows and saltwater marshes, and in Italy it grows along the coasts. The genus Inula for its richness in secondary metabolites has been the subject of many recent researche’ studies, which highlighted the different bioactivities of several extracts obtained from this plant, due to different secondary metabolites content. Some Inula spp. contain interesting compounds with antibacterial, antifungal, cytotoxic and antioxidant activities (Mosaad et al. 2008). In a recent paper, *Corresponding author. Email: [email protected] This manuscript was submitted at SIF 2013, XIII National Congress of Societa` Italiana di Fitochimica (Italian Phytochemical Society), Gargnano (Italy), 19 –21 September 2013. q 2014 Taylor & Francis
Downloaded by [Nova Southeastern University] at 08:25 06 January 2015
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A. Bucchini et al.
Omezzine et al. (2011) showed a good antifungal activity of different Inula extracts against two Trichoderma species and three formae speciales of Fusarium oxysporum. Furthermore, Giamperi et al. (2010) showed a good antioxidant activity of the essential oil obtained from I. crithmoides aerial parts and a considerable antifungal activity of extracts obtained from callus cultures. Fungi and bacteria have unfavourable effects on quality, safety and preservation of food. Synthetic chemicals are widely used in the control of plant diseases; however, these compounds may leave toxic residues in treated products (Zhao et al. 2010). Therefore, there is an urgent demand for the discovery of new safer and more effective antimicrobial substances, and medicinal plants can be regarded as a valid and alternative source of bioactive compounds (Bucchini et al. 2013). Among these, the components with phenolic structures such as carvacrol, eugenol and thymol were highly active against pathogens and showed a good antioxidant activity. The aim of our work was to evaluate the total phenolic content, antioxidant and antifungal activities of three different extracts obtained from I. crithmoides aerial parts.
2. Results and discussion 2.1. Total phenolic content Powdered dried aerial parts were sequentially extracted by maceration with n-hexane (H), methylene chloride (DCM) and methanol (MeOH). Yields of extractions are reported in Table S1. Looking at the same table it is evident that among the three Inula extracts, the methanol extract showed the highest total phenolic content, 15.52 mg/g dry weight of starting material (DW). Furthermore, the total polyphenols content exhibited the descending order among MeOH . H . DCM extracts.
2.2. Free-radical scavenging activity The DPPH radical assay has been widely used to test the ability of compounds as free-radical scavengers or hydrogen donors and to evaluate the antioxidative activity of plant extracts and foods (Prior et al. 2005; Giamperi et al. 2012; Seow et al. 2012). Our results (Table S1) show that the methanol and hexane extracts exhibited the highest DPPH-radical scavenging activity with EC50 value of 0.59 and 0.57 mg DW/mL, respectively. These EC50 values are six times higher than that of the ascorbic acid (0.110 mg DW/mL). Antioxidant activity assessed with the DPPH test in both extracts is linearly related with polyphenols content (R 2 is 0.99 for methanol extract and is 0.97 for hexane extract).
2.3. Anti- inflammatory activity (inhibition of lipid peroxidation) 5-Lipoxygenase (5-LOX) is one of the key enzymes in the synthesis of leukotrienes, inflammatory eicosanoids and histamine from mast cells, and in the formation of cytokines (mainly TNF-a, IL-6) by macrophages followed by the liberation ROS (reactive oxygen species). Thus, a reduction of the production of inflammatory mediators by inhibiting the activities of 5-lipoxygenase or cyclooxygenase in addition to a direct ROS scavenging by antioxidants is considered as a promising therapeutic target (El-Readi et al. 2013). The aim of this step of our work was to evaluate the inhibitory effect of different I. crithmoides extracts on 5-lipoxygenase. The hexane extract showed greater inhibitory effect with an IC50 similar to that of Trolox and ascorbic acid (IC50 ¼ 13.48, 11.89 and 18.63 mg DW/ mL, respectively) (Table S1).
Downloaded by [Nova Southeastern University] at 08:25 06 January 2015
Natural Product Research
3
2.4. Antifungal activity Recently, in different parts of the world, attention has been paid towards exploitation of higher plant products as novel chemotherapeutants in plant protection (Kotan et al. 2010). For this reason we decided to test the obtained extracts for their antifungal effect against eight phytopathogenic fungi and their possible activity in control of fungal diseases. The antifungal activity was tested in vitro by using the agar dilution method. Extracts demonstrated a good antifungal activity against Phytophtora cryptogea and Alternaria solani. Particularly, the antifungal activity of methanol extract at the dose of 400 ppm was comparable with the activity of the positive control (Nystatin 50 ppm). Also the hexane extract exhibited a good antifungal activity. In this case the dose of 400 ppm of the extract showed an important antifungal activity when compared with the untreated control with a similar trend of the methanol extract. A weaker activity was shown against all fungi of the genus Fusarium. Only for F. oxysporum, the fungicidal activity of extracts was similar to that shown towards A. solani (Tables S2 –S4). The DCM extract is not effective against all fungi tested (data not shown).
3. Conclusions The present report is the first comprehensive study of the polyphenolic content, antioxidant, anti-inflammatory and antifungal activities of I. crithmoides extracts. Interestingly, the antiinflammatory activity of I. critmoides methanol and hexane extracts positively correlated with their polyphenolic content (R 2 ¼ 0.94 and 0.95, respectively). This study indicated that the methanolic and the hexane extracts of I. crithmoides might be regarded as natural antioxidant and anti-inflammatory agents. In agriculture, these extracts might be applicable as natural chemicals for integrated plant protection against a variety of phytopathogenic fungi that cause an economically important tomato and potato disease. Further studies are required to investigate the in vivo efficacy. With efficient collaborations with pharmacologists and medical doctors, plant pathologists will be crucial to see the development of a comprehensive study.
Supplementary material Experimental details relating to this paper are available online, alongside Tables S1 – S4.
References Bucchini A, Giamperi L, Ricci D. 2013. Total polyphenol content, in vitro antifungal and antioxidant activities of callus cultures from Inula crithmoides L. Nat Prod Commun. 8:1587–1590. El-Readi MZ, Eid HH, Ashour ML, Eid SY, Labib RM, Sporer F, Wink M. 2013. Variations of the chemical composition and bioactivity of essential oils from leaves and stems of Liquidambar styraciflua (Altingiacae). J Pharm Pharmacol. 65:1653 –1663. Giamperi L, Bucchini A, Bisio A, Giacomelli E, Romussi G, Ricci D. 2012. Total phenolic content and antioxidant activity of Salvia spp. exudates. Nat Prod Commun. 7:201–202. Giamperi L, Bucchini A, Fraternale D, Genovese S, Curini M, Ricci D. 2010. Composition and antioxidant activity of Inula crithmoides essential oil grown in central Italy (Marche Region). Nat Prod Commun. 5:315–318. Kotan R, Cakir A, Dadasoglu F, Aydin T, Cakmakci R, Ozer H, Kordali S, Mete E, Dikbas N. 2010. Antibacterial activities of essential oils and extracts of Turkish Achillea, Satureja and Thymus species against plant pathogenic bacteria. J Sci Food Agric. 90:145–160. Mosaad A, Abdel-Wahhab S, Abdel-Azim H, Aziza AEN. 2008. Inula crithmoides extract protects against ochratoxin Ainduced oxidative stress, clastogenic and mutagenic alteration in male rats. Toxicon. 52:566–573. Omezzine F, Daami-Remadi M, Rinez A, Ladhari A, Haoulala R. 2011. In vitro assessment of Inula spp. organic extracts for their antifungal activity against some pathogenic and antagonistic fungi. Afr J Microbiol Res. 5:3527–3531.
4
A. Bucchini et al.
Downloaded by [Nova Southeastern University] at 08:25 06 January 2015
Prior RL, Wu X, Schaich K. 2005. Standardized methods for the determination of antioxidant capacity and phenolic in foods and dietary supplements. J Agric Food Chem. 53:4290–4302. Seow MH, Amru NB, Chandran S. 2012. Antioxidant activity, phenolic and flavonoid contents in the leaves of different varieties of sweet potato (Ipomoea batatas). Aust J Crop Sci. 6:375–380. Zhao J, Li Y, Liu Q, Gao K. 2010. Antimicrobial activities of some thymol derivatives from the roots of Inula hupehensis. Food Chem. 120:512–516.
