Hydrodistilled leaves of Chenopodium album yielded 0.64 % v/w of essential oil. GC and
GC/MS analyses of the oil revealed that the bulk of the oil was constituted by aromatic compounds (60.1 %). The abundant constituents of the oil were: p- cymene (40.9 %), ascaridole (15.5 %), pinane-2-ol (9.9 %), α-pinene (7.0 %), β-pinene (6.2 %) and α-terpineol (6.2 %). The oil displayed strong anti-inflammatory activity against 12-O-tetradecanoylphorbol-13-acetate (TPA) – induced ear edema in mice

Edewor, T. I.

Hamid, A. A.

Muhammad, N. O.

Olawore, N. O.

Saliu, B. K.

Usman, L. A.

Eldorado - Ressourcen aus und für Lehre, Studium und Forschung

EXCLI Journal 2010;9:181-186 – ISSN 1611-2156 Received: August 15, accepted: December 04, 2010, published: December 07, 2010  181 Original article: CHEMICAL CONSTITUENTS AND ANTI-INFLAMMATORY ACTIVITY OF LEAF ESSENTIAL OIL OF NIGERIAN GROWN Chenopodium album L.  L.A. Usman*1, A.A. Hamid1, N.O. Muhammad2, N.O. Olawore3, T.I. Edewor3, B.K. Saliu4  1 Department of Chemistry, University of Ilorin, Ilorin, Nigeria 2 Department of Biochemistry, University of Ilorin, Ilorin, Nigeria 3 Department of Pure and Applied Chemistry, Ladoke Akintola University of Technology, Ogbomoso, Nigeria  4 Department of Microbiology, University of Ilorin, Ilorin, Nigeria  * Corresponding author’s email Address: lamidiusman09@gmail.com; usmanlami-di@yahoo.com   ABSTRACT Hydrodistilled leaves of Chenopodium album yielded 0.64 % v/w of essential oil. GC and GC/MS analyses of the oil revealed that the bulk of the oil was constituted by aromatic com-pounds (60.1 %). The abundant constituents of the oil were: p- cymene (40.9 %), ascaridole (15.5 %), pinane-2-ol (9.9 %), α-pinene (7.0 %), β-pinene (6.2 %) and α-terpineol (6.2 %). The oil displayed strong anti-inflammatory activity against 12-O-tetradecanoylphorbol-13-acetate (TPA) – induced ear edema in mice.   Keywords: Chenopodium album; chenopodiaceae; p-cymene; ascaridole; pinane-2-ol;  α-terpineol; anti-inflammatory    INTRODUCTION Chenopodium album L. (family Cheno-podiaceae) is an annual shrub widely grown in Europe, North America, Asia, and Af-rica. It is commonly known as pigweed, fat hen or lamb-quarters’ (Bailey, 1977; GRIN Database, 2005). The plant is used in folk medicine in different parts of the world. As therapeutic agents, it is used as laxative, antihelmintic against round and hook worms and blood purifier. Its use for the treatments of hepatic disorders, spleen en-largement, intestinal ulcers and burns has also been documented (Sarma et al., 2008). Various bioactivities such as antifungal (Tahara et al., 1994 Maruta et al., 1995), antipruritic, antinociceptive (Dai et al., 2002) and hypotensive (Gohar and Elma-zar, 1997) properties of crude and isolated compounds from the plant justified its uses in traditional medicine. Phytochemical ana-lyses revealed the presence of alkaloids (Horio et al., 1993; Cutillo et al., 2004), apocarotenoids (DellaGreca et al., 2004), flavonoids (Gohar and Elmazar, 1997), phytoecdysteroids (Dinan, 1992; Dinan et al., 1998; DellaGreca et al., 2005a) and an unusual xyloside (DellaGreca et al., 2005b) in the plant. Earlier work on the leaf essen-tial oil of East Mediterranean grown C. al-bum revealed the abundance of Limonene (23.2 %), α-terpinyl acetate (13.7 %), α-terpinene (12.3 %) and cis-ascaridole (12.2 %) in the oil (Dembitsky et al., 2008). It has been established that geographical and agroclimatic conditions could affect the composition pattern of essential oil of a par-EXCLI Journal 2010;9:181-186 – ISSN 1611-2156 Received: August 15, accepted: December 04, 2010, published: December 07, 2010  182 ticular plant and therefore its biological ac-tivities (Lahlou, 2004). Based on these fac-tors, we investigate the leaf essential oil of Nigerian grown C. album and its anti-inflammatory activity.  EXPERIMENTAL  Plant materials The fresh leaves of Chenopodium al-bum were obtained in Ilorin, Kwara State, North Central, Nigera. Identification was carried out at the herbarium of the Depart-ment of Plant Biology, University of Ilorin, Nigeria, where voucher specimens were deposited.  Oil isolation Pulverized leaves of Chenopodium al-bum were hydrodistilled for 3 h in a Clevenger-type apparatus according to the British Pharmacopoeia (1980) specification. The resulting oil was collected, preserved in a sealed sample tube and stored under re-frigeration until analysis.  Gas chromatography GC analysis were performed on an orion micromat 412 double focusing gas chromatography system fitted with two cap-illary columns coated with CP-Sil 5 and CP-Sil 19 (fused silica, 25 m×0.25 mm, 0.15 µm film thickness) and flame ioniza-tion detector (FID). The volume injected was 0.2 µL and the split ratio was 1:30. Oven temperature was programmed from 50–230° C, respectively. Qualitative data were obtained by electronic integration of FID area percents without the use of correc-tion factors.  Gas chromatography/mass spectrometry A Hewlett Packard (HP 5890A) GC in-terfaced with a VG Analytical 70-250S double focusing mass spectrometer was used. Helium was the carrier gas at 1.2 ml/min. The MS operating conditions were: ionization voltage 70 ev, ion source temperature 230° C. The GC was fitted with a 25 m×0.25 mm, fused silica capillary column coated with CP-Sil 5. The film thickness was 0.15 µm. The GC operating conditions were identical with those of GC analysis. The MS data were acquired and processed by online desktop computer equipped with disk memory. The percent-age compositions of the oil were computed in each case from GC peak areas. The iden-tification of the components was based on the retention indices (determined relative to the retention times of series of n-alkanes) and mass spectra with those of authentic samples and with data from literature (Jennings and Shibamito, 1980; Adams 1995; Joulain and Koenig, 1998).  Anti-inflammatory activity of leaf essential oil of C. album The animal experiment were approved by the Department of Biochemistry, Uni-versity of Ilorin, Animal Care and Use Committee and conducted according to standard guidelines. Sixty-four, 21-day old male swiss Webster mice, weighing be-tween 22 and 25 g were housed in groups of 8 in an NIH – approved facility. All groups were fed with standard rodent diet (Test diet 570B, Purina Mills, St. Louis, MO) and supplied water ad libitum, throughout the experimental period. The light in the facil-ity were turned off between 1900 and 0700 h, with the environmental temperature maintained at 24 ± 1° C. All experimental procedures were conformed to the National Institute of Health, Public Health Service and Animal Welfare Act guidelines for the ethical treatment of laboratory animals.  Topical anti-inflammatory assay A modification of the method of Young et al. (1984) was used. The topical anti-inflammatory activity was evaluated as in-hibition of the 12-O-tetradecanoylphorbol-13-acetate (TPA) induced ear edema in mice by the oils following standard proce-dure (Young et al., 1984; Griffiths et al., 1988; Bralley et al., 2008). Edema was in-duced in ears of each mouse by the topical application of 2 µg TPA dissolved in 20 µl of acetone to both the inner and outer sur-EXCLI Journal 2010;9:181-186 – ISSN 1611-2156 Received: August 15, accepted: December 04, 2010, published: December 07, 2010  183 faces of the right ear (surface: about 1 cm2). Thirty minutes after the application of TPA, the inner and outer surfaces of each ear were treated (10 ul to each side) with: 1. 50 % ethanolic solutions of the test es-sential oil (EO) in doses of 0.625 (CA4), 1.25 (CA3), 2.5 (CA2) and 5.0 (CA1) mg eo/ear (n = 8 at each dosage) 2. 50 % ethanol (vehicle control) 3. Indomethacin (0.25 mg/ear dissolved in 50 % ethanol (as a standard anti-inflammatory drug). The thickness of each ear was measured using a micrometer (Mitutoyo Series IP65, Mitutoyo America, Aurora, IL) before and at 4 h and 24 h after tetradecanoylphorobol-13-acetate administration. The micrometer was applied near the top of the ear distal to the cartilaginous ridges. At 24 h, each ani-mal was sacrificed and a plug biopsies (6 mm diameter hole punch) were removed from both the treated (right) and the un-treated (left) ears immediately, weighed, frozen and stored at 8° C. The edematous response was measured as the weight dif-ference between the two plugs. The anti-inflammatory activity was expressed as percentage of the edema reduction in treated mice compared to the control mice. The pharmacological data were analyzed by the Student’s t-test, and a probability level lower than 0.05 was considered as signifi-cant. RESULTS AND DISCUSSION Pulverized leaves of Chenopodium al-bum afforded oil in the yield of 0.64 % v/w. Table 1 shows the retention indices, mass spectra data, identities and relative percent-age of the constituents of the oil. A total of 37 compounds that represent 95.6 % of the oil were identified from their mass spectra. Hydrocarbon and oxygenated monoterpenes constituted 17.4 and 18.1 % of the oil. The percentage composition of aromatic com-pounds in the oil was 60.9 %. The oil was characterized by the abun-dance of aromatic compounds. Bulk of the aromatic compounds was constituted by p-cymene (40.9 %). Other notable aromatic compounds in the oil were ascaridole (15.5 %) and ethyl cinnamate (3.7 %). α-pinene was the most abundant hydro-carbon monoterpene in the oil. Other hy-drocarbon monoterpenes that were found in significant proportions were β-pinene (6.2 %), limonene (4.2 %). The most abun-dant oxygenated monoterpene in the oil was pinane-2-ol (9.9 %). α-terpineol (6.2 %) and linalyl acetate (2.0 %) also occurred in ap-preciable quantities. Qualitative and quantitative composi-tions of the oil were found to be different from the leaf essential oil of East Mediter-ranean grown C. album (Dembitsky et al., 2008). For instance, the principal constitu-ents in the leaf oil of the Nigerian grown species, α-terpineol, pinane-2-ol, linalyl acetate, p-cymene and ethyl cinnamate, were not identified in the oil of East Medi-terranean C. album. The most abundant constituent of the oil was p-cymene; hence, the oil was of p-cymene chemotype. On the other hand, terpinenyl acetate and terpinen-1-ol that were found in appreciable amounts in the oil of East Mediterranean grown C. album were not identified in the oil of the Nigerian grown C. album. Meanwhile, α-pinene, β-pinene, limonene and ascaridole were identified in both oils. These constitu-ents were of greater abundance in the oil of the Nigerian grown C. album except limo-nene that predominate the leaf oil of East Mediterranean grown C. album. Variations in composition patterns of the oils may be attributed to agroclimatic and geographical conditions. It has been established that anti-inflammatory activities of essential oils are attributable to the presence of substituent such as; limonene, linalool, linalyl acetate and α-pinene (Peana et al., 2002; Özbek and Sever, 2005; Karaca et al., 2007).   EXCLI Journal 2010;9:181-186 – ISSN 1611-2156 Received: August 15, accepted: December 04, 2010, published: December 07, 2010  184 Table 1: Chemical composition (%) of leaf oil of Chenopodium album Compounda RIb  Percentage  composition Mass spectra data Tricyclene  922 Tr 136, 121, 105, 93 α-thujene 926 Tr 136, 121, 115, 105 α-pinene 933 7.0 136, 121, 105, 93 Camphene 946 Tr 136, 121, 107, 93 Sabinene 971 Tr 136, 121, 107, 93 β-pinene  976 6.2 136, 121, 107, 93 Myrecene 990 Tr 136, 121, 115, 107 p-cymene 1022 40.9 134, 119, 103, 91 Limonene 1027 4.2 136, 121, 107, 93 Benzyl alcohol 1028 Tr 108, 91, 89, 79 1,8-cineole 1029 Tr 154, 139, 125, 108 Cis-ocimene 1035 Tr 136, 121, 105, 93 γ-terpinene 1057 Tr 136, 121, 105, 93 Linalool 1098 Tr 139, 121, 109, 97 Pinane-2-ol 1136 9.9 139, 111, 93, 69 Allo ocimene 1142 Tr 136, 121, 105, 93 Citronellal 1150 Tr 154, 136, 121, 111 Borneol 1162 Tr 139, 121, 110, 95 Terpinen-4-ol 1175 Tr 154, 136, 71, 43 α-terpineol 1188 6.2 136, 121, 105, 93 Citronellol 1226 Tr 138, 123, 109, 99 Ascaridole 1237 15.5 139, 125, 69, 41 Neral 1238 Tr 135, 119, 109, 99 Linalyl acetate 1255 2.0 136, 121, 105, 93 Geranial  1268 Tr 152, 137, 123, 109 Borneol acetate 1284 Tr 196, 154, 136, 108 Thymol 1290 Tr 150, 135, 121, 107 Carvacrol 1299 Tr 150, 135, 121, 107 Ethyl cinnamate 1460 3.7 176, 158, 147, 131 Acetyl eugenol 1523 Tr 207, 164, 149, 121 Elemicin 1553 Tr 208, 193, 177, 150 Benzyl benzoate 1759 Tr 152, 105, 91, 51 Total  95.6  a Compounds are listed in order of elution from Silica Capillary Column coated on CP-Sil 5 b Retention indices on fused Silica Capillary Column coated with CP-Sil 5=trace (<0.1 %)  EXCLI Journal 2010;9:181-186 – ISSN 1611-2156 Received: August 15, accepted: December 04, 2010, published: December 07, 2010  185 The result of anti-inflammatory activity of leaf essential oil of Chenopodium album is presented in Table 2.The result revealed that the anti-inflammatory action of the oil is concentration dependent. Hence, the per-centage reduction in the ear edema in-creases with increase in concentration of the oil. Furthermore, the oil caused signifi-cant reduction (p < 0.05) in the ear edema except at 0.625 mg concentration. The re-duction may be due to the presence of α-pinene, linalool and linaly acetate in the oil. Hence, the oil can be use as anti–inflammatory agent.  Table 2: Anti-inflammatory activity of leaf  essential oil of Chenopodium album   P < 0.05, Student’s t-test CA = Chenopodium album  REFERENCES Adams RP. Identification of essential oil components by gas chromatography and mass spectrometry. Carol Stream, IL.: Al-lured Publ. Corp., 1995. Bailey LH. Manual of cultivated plants. Most commonly grown in the continental United States and Canada. Rev. ed., compl. restudied. NewYork: Macmillan, 1977. Bralley EE, Greenspan P, Hargrove JL, Wicker L, Hartle DK. Topical anti-inflammatory activity of Polygonum cuspi-datum extract in the TPA model mouse ear inflammation. J Inflamm 2008;5:1. British Pharmacopoeia II. 109. London: HMSO, 1980. Cutillo F, D’Abrosca B, DellaGreca M, Zarrelli A. Chenoalbicin, a novel cinnamic acid amide alkaloid from Chenopodium al-bum. Chemistry and Biodiversity 2004;1: 1529-83. Dai Y, Ye WC, Wang ZT, Matsuda H, Kubo M, But PPH. Antipruritic and antino-ciceptive effects of Chenopodium album L. in mice. J Ethnopharmacol 2002;81:245-50. DellaGreca M, Di Mariono C, Zarrelli A, D’Abrosca B. Isolation and phytotoxicity of apocarotenoids from Chenopodium al-bum. J Nat Prod 2004;67:1492-5. DellaGreca M, D’Abrosca B, Fiorentino A, Previtera L, Zarrelli A. Structure elucida-tion and phytotoxicity of ecdysteriods from Chenopodium album. Chemistry and Bio-diversity 2005a;2:457-62. DellaGreca M, Previtera L, Zarrelli A. A new xyloside from Chenopodium album. Nat Prod Res 2005b;19:87-90. Dembitsky V, Shkrob I, Hanus LO. Ascari-dole and related peroxides from the genus Chenopodium. Biomed Pap Med Fac Univ Palacky Olomuc Czech Repub 2008;152: 209-15. Dinan L. The analysis of phytoecdysteroids in single (preflowering stage) specimens of fat hen, Chenopodium album. Phytochem Anal 1992;3:132-8.  Dinan L, Whiting P, Scott AJ. Taxonomic distribution of phytoecdysteroids in seeds of members of the chenopodiaceae. Bio-chem Syst Ecol 1998;26:553-76. Group Dose mg No of animals in the group Change in ear weight (Mean ±SEM) mg Percent-age of edema reduc-tion Control - 8 7.20 ± 0.02 - Indo-methacin 0.25 8 3.10 ± 0.01 57.00 CA1 5.00 8 0.50 ± 0.02 91.80 CA2 2.50 8 0.70 ± 0.05 89.40 CA3 1.25 8 1.90 ± 0.04 74.40 CA4 0.625 8 4.60 ± 0.01 41.00 EXCLI Journal 2010;9:181-186 – ISSN 1611-2156 Received: August 15, accepted: December 04, 2010, published: December 07, 2010  186 Gohar AA, Elmazar MMA. Isolation of hy-potensive flavonoids from Chenopodium species growing in Egypt. Phytother Res 1997;11:564-7. Griffiths RJ, Wood BE, Li S, Blackham A. Pharmacological modification of 12-O-tetradecanoylphorbol-13-acetate induced inflammation and epidermal cell prolifera-tion in mouse skin. Agent Actions 1988;25: 344–51. GRIN Database USDA, ARS. National Ge-netic Resources Program, Germplasm Re-sources Information Network (GRIN), Na-tional Germplasm Research Laboratory, Beltsville, Maryland. Available online at http://www.ars-grin.gov2/cgi-bin/npgs/html/genform.pl 2005. Horio T, Yoshida K, Kikuchi H, Kawabata J, Mizutani J. A phenolicamide from roots of Chenopodium album. Phytochemistry 1993;33:807-8. Jennings W, Shibamito I. Qualitative analy-sis of flavour volatiles by gas capillary chromatography. New York: Academic Press, 1980. Joulain D, Koenig WA. The atlas of spectra data of sequiterpene hydrocarbons. Ham-burg: E.B. Verlag, 1998. Karaca M, Özbek H, Him A, Tütüncü M, Akkan HA, Kaplanoğlu V. Investigation of anti-inflammatory activity of Bergamot oil. Eur J Gen Med 2007;4:176–9. Lahlou M. Methods to study the phyto-chemistry and bioactivity of essential oils. Phytother Res 2004;18:435–48. Maruta Y, Fukushi Y, Onkawa K, Naka-nishi Y, Tahara S, Mizutani J. Antimicro-bial stress compounds from Hypochoeris radicata. Phytochemistry 1995;38:1169–73. Özbek H, Sever B. Investigation of lethal doses and anti-inflammatory effect of al-pha-pinene on mice and rats. Turkish Phar-macological Society, 18th National Phar-macology Congress, September, 2005, Iz-mir-Turkey, P-118. Peana AT, D’Aquila PS, Panin F, Serra G, Pippia P, Moretti, MD. Anti-inflammatory activity of linalool and linalyl acetate con-stituents of essential oils. Phytomedicine 2002;9:721–6. Sarma H, Sarma A, Sarma CM. Traditional knowledge of weeds: a study of herbal medicines and vegetables used by the As-samese people (India). Kerba Polnica 2008; 54:80–8. Tahara S, Kassai S, Innoue M, Kawabata J, Mizutani J. Identification of mucondialde-hyde as a novel stress metabolite. Experien-tia 1994;50:137-41. Young JM, Spires DA, Bedford CJ, Wagner B, Ballaron SJ, De Young LM. The mouse ear inflammatory response to topical ara-chidonic acid. J Invest Dermatol 1984;82: 367–71. 

Chemical constituents and anti-inflammatory activity of leaf essential oil of Nigerian grown Chenopodium album L.

https://eldorado.tu-dortmund.de/bitstream/2003/27739/1/Usman_071210_proof.pdf

Chemical constituents and anti-inflammatory activity of leaf essential oil of Nigerian grown Chenopodium album L.

Abstract

Similar works

Full text

Available Versions

Eldorado - Ressourcen aus und für Lehre, Studium und Forschung