Printer Friendly

Analysis and Phytotoxicity of Oily Fraction of Aerial Parts of Cichorium intybus.

Byline: SHAZIA SHAH, AZHAR MAHMOOD, SUMAYYA SAIED AND ABDUL MALIK

Summary: The oil obtained from n-hexane soluble sub-fraction of the aerial parts of Cichorium intybus was subjected to GC and GC-MS analysis, revealing the presence of thirty five components which were identified by using NIST mass spectral search program and further confirmed by their Kovat's retention indices. The oil showed significant phytotoxic activity (61.12% inhibition against Lemna minor at high dose of 1000 (mu)g/ml).

Keywords: Cichorium intybus, Asteraceae, Oil Composition, GC, GC-MS.

Introduction

The family Asteraceae comprises about 1535 genera and 23000 species. In Pakistan this family is represented by over 650 species [1]. Cichorium intybus (commonly known as Wild Endive) is small to medium size perennial herb up to 30 cm in height, with a flashy taproot up to 75 cm in length [2, 3]. Its roots are carminative, diuretic, hypoglycemic and effective in jaundice, liver enlargement, gout, and rheumatism. The roots and leaves cure inflammatory swellings [4]. The whole plant is bitter, acrid, thermogenic, anti-inflammatory, appetizer, digestive, stomachic, alexeferic and tonic [5]. Previous phytochemical investigations of Cichorium intybus have resulted in the isolation and identification of a large variety of compounds including anthocyanins [6], sesquiterpene lactones [7], fructans [8], flavonoids [9] and coumarins [10]. However, the composition of oil has not so far been determined.

We now report the analysis of the oil of C. intybus by a combination of gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS).

Results and discussion

The ethanolic extract of C. intybus was fractionated into n-hexane, ethyl acetate, n-butanol and water soluble sub-fractions. Out of these the hexane soluble sub fraction exhibited phytotoxicity. On further workup of the n-hexane soluble fraction, the major phytotoxicity was located in its oily fraction. This prompted us to carry out further studies on this fraction.

Both qualitative and quantitative assessments of the oily fraction were carried out by GC and GC-MS techniques. The GC chromatogram exhibited forty one components, (Fig. 1), out of which thirty five have been identified by their respective mass fragmentations. It was further confirmed by calculation and comparison of their retention indices with those cited in literature [11-23]. All of these are being reported for the first time from this plant. The percentage contents and calculated retention indices of the constituents are summarized in Table-1. Resultant compounds could mainly be categorized into three groups namely, benzene derivatives (21 compounds), long chain esters/acids (12 compounds) and long chain alkanes (02 compounds). These compounds together accounted for 97.89% of oil.

Phytotoxicity is used to describe the degree of toxic effect by a compound or compound fraction on plant growth. Such damage may be caused by a wide variety of compounds, including trace metals, pesticides, salinity, phytotoxins or allelopathy. The phytotoxicity of a variety of oils and essential oils recently reported in literature prompted us to determine the phytotoxicity of the oily fraction. It was carried out by the method described in the experimental. The oil showed significant phytotoxic activity (61.12% inhibition against Lemna minor at high dose of 1000 (mu)g/ml) (Table-2).

Table-1: GC and GC-MS analysis of oil of aerial parts of Cichorium intybus.

No.###Compounds identified###Rt(Min)###RI###%Qty###Some important peaks###m/z (% abundance)

1.###1-Methoxy,4-(1-propenyl) benzene###12.5###1266###0.07###148(100), 133(24), 117(23), 105(22), 91(16), 77(18).

2.###2-Methyl, 5-(1-methylethyl) phenol###13.7###1296###0.10###150(22), 135(100), 117(11), 107(6), 91(12), 77(5).

3.###3-Allyl, 6-methoxyphenol###14.9###1358###0.15###164(100), 149(36), 137(18), 131(23), 103(23), 91(18), 77(21).

4.###1-Butylhexyl benzene###19.9###1530###2.52###218(9), 161(12), 147(20), 105(10), 91(100).

5.###1-Propylheptyl benzene###20.2###1536###2.74###218(9), 175(13), 133(21), 119(5), 105(8), 91(100).

6.###1-Ethyloctyl benzene###20.7###1554###2.56###218(18), 189(19), 147(3), 133(5), 119(52), 105(15), 91(100), 77(3).

7.###1-Methylnony lbenzene###21.8###1592###3.13###218(8), 119(2), 105(100), 91(12), 77(4).

8.###1-Butylheptyl benzene###22.3###1624###3.38###232(12), 175(15), 161(3), 147(21), 119(5), 105(14), 91(100), 77(2).

9.###1-Propyloctyl benzene###22.9###1636###4.05###232(11), 189(14), 147(2), 133(32), 119(5), 105(12), 91(100), 77(2).

10.###1-Ethylnonyl benzene###23.2###1658###4.79###232(13), 203(19), 161(2), 147(4), 119(60), 105(13), 91(100), 77(2).

11.###1-Methyldecyl benzene###24.8###1694###4.09###232(13), 119(3), 105(100), 91(12), 79(5), 71(4).

12.###1-Pentylheptyl benzene###25.5###1720###1.97###246(12), 175(18), 161(21), 133(3), 119(9), 105(19), 91(100), 77(3).

13.###1-Butyloctyl benzene###25.6###1724###3.32###246(15), 189(17), 161(3), 147(26), 133(5), 119(7), 105(17), 91(100), 77(3).

14.###1-Propylnonyl benzene###25.9###1732###1.77###246(9), 203(12), 161(3), 147(4), 133(32), 119(5), 105(10), 91(100), 77(3).

15.###1-Ethyldecyl benzene###26.5###1792###1.88###246(10), 217(20), 161(4), 147(5), 133(6), 119(63), 105(13), 91(100), 77(3).

16.###1-Methylundecyl benzene###27.6###1795###2.85###246(10), 231(2), 147(2), 117(3), 105(100), 91(9), 79(4).

17.###1-Pentyloctyl benzene###28.1###1813###0.32###260(12), 189(10), 175(10), 161(14), 147(3), 133(5), 119(10), 105(20), 91(100), 77(4).

18.###1-Butylnonyl benzene###28.2###1822###0.21###260(10), 203(12), 161(3), 147(30), 119(7), 105(22), 91(100), 83(3).

19.###1-Propyldecyl benzene###28.5###1836###0.65###260(8), 217(10), 161(4), 133(35), 119(8), 105(12), 91(100), 85(8).

20.###1-Ethylundecyl benzene###29.2###1860###0.27###260(16), 231(20), 189(2), 147(8), 133(8), 119(61), 105(17), 91(100), 85(4).

21.###Nonadecane###29.8###1900###0.62###268(5), 197(4), 183(4), 169(7), 99(18), 85(58), 71(83), 57(100).

22.###1-Methyldodecyl benzene###30.1###1904###0.53###260(9), 245(2), 105(100), 91(10), 79(4).

23.###Methyl hexadecanoate###30.6###1916###0.29###270(22), 239(14), 227(20), 213(4), 199(8), 185(9), 143(22), 87(76), 74(100).

24.###Methyl 2-methylhexadecanoate###32.5###1968###12.32###284(28), 227(5), 213(10), 199(9), 171(5), 157(32), 115(12), 101(98), 88(100).

25.###Hexadecanoic acid###33.6###1974###5.97###256(56), 227(8), 213(40), 185(18), 171(19), 157(20), 129(59), 83(28), 73(100).

26.###Ethyl linoleate###36.5###2156###6.18###308(10), 263(20), 237(4), 191(6), 178(6), 109(42), 95(98), 81(89), 67(100).

27.###Ethyl linolenate###36.6###2174###11.74###306(20), 277(5), 261(14), 250(12), 191(6), 135(24), 121(37), 108(57), 95(88), 79(100)

28.###Octadecanoic acid###38.0###2182###9.04###284(72), 255(8), 241(42), 227(12), 185(40), 129(60), 97(52), 83(50), 73(100).

29.###Docosanoic acid###41.3###2376###1.62###340(24), 311(6), 297(20), 241(12), 157(24), 129(48), 101(81).

30.###Methyl 2, 4-dimethylheneicosanoate###45.1###2613###2.80###368(42), 325(22), 297(4), 199(8), 129(6), 111(8), 101(80), 88(100).

31.###Methyl tetracosanoate###46.9###2714###1.97###382(20), 393(10), 297(4), 283(4), 199(5), 157(14), 143(8), 129(8), 111(12).

32.###Methyl pentacosanoate###48.7###2819###1.88###396(52), 353(28), 311(6), 297(10), 213(8), 199(7), 185(5), 157(37), 143(11).

###Methyl 10,14,18,22-

33.###tetramethyltricosanoate###52.1###3022###0.24###424(32), 409(8), 381(16), 339(7), 199(9), 143(14), 97(44), 57(100).

34.###Hentriacontane###53.9###3100###1.68###436(2), 393(4), 295(5), 197(8), 155(12), 127(18), 99(32), 85(66), 71(82), 57(100).

35.###Methyl 2, 4, 6-trimethyl

###hexacosanoate###56.2###3218###0.19###452(78), 381(10), 365(8), 281(12), 129(30), 111(44), 101(84), 88(86).

###Retention time in Minutes,###Kovat's Retention Indices

Table-2: In vitro phytotoxic activity of oil of C. intybus against Lemna minor.

###Conc. ((mu)g/ml)###No. of Fronds###% Inhibition

Sample Oil###-ve Control###Sample###-ve Control

1000###07###61.12

100###0.015###11###18###38.89

10###15###16.67

Experimental

The aerial parts of Cichorium intybus L. were collected from Gilgit, Pakistan in July, 2007 and identified by the Plant Taxonomist at the Department of Botany, University of Karachi, where a voucher specimen (No. 1036, general herbarium # 71320) has been deposited.

Extraction

The shade dried aerial parts of Cichorium intybus (9.5 kg) were ground and extracted with 95% ethanol (3 x30 L) for a period of 7 days each at room temperature. The combined ethanolic extract (418 g) was divided into n-hexane (190 g), ethyl acetate (45 g), n-butanol (132 g) and water soluble (40 g) sub- fractions. The n-hexane soluble sub-fraction was dissolved in n-hexane and adsorbed over silica gel 60 (70-230 mesh size, E-Merck). Elution with n-hexane provided the crude turbid oil. It was charcoaled and filtered with celite to furnish light yellow transparent oil. (30.56 g) which was analyzed by GC and GC- MS, resulting in the separation and subsequent identification of thirty five compounds.

GC and GC-MS analysis

The GC was performed on a Shimadzu-GC-9A gas chromatograph, FID at 220, N2 at 1.0 ml/min , SPB-5 capillary column (30 m x 0.53 mm ID; 0.3 (mu)df); split ratio 1:30, injector temperature was 240 degC, the column temperature was maintained at 50 degC for the first 5 min and then raised to 235 degC (5degC/min). GC-MS were carried out Hewlett-Packard 5890 gas chromatograph, combined with a Jeol, JMS-HX 110 mass spectrometer with source at 270 oC with splitting ratio 1:30. The analyses were performed on the aforementioned program on an equivalent column HP-5 (25 m x 0.22 mm and 0.25 (mu)mdf).

Identification of compounds

The resolved components were characterized by a mass spectral survey using the NIST mass spectral search program and GC-MS Library. Their identification was further authenticated through comparison of their respective Kovat's retention indices (RI) cited in literature [11-23].

Phytotoxicity

For phytotoxic assay, a sample solution was prepared by dissolving 30 mg of oil fraction into 1.5 ml of Ethanol. Three flasks were inoculated with 10, 100 and 1000 (mu)l of sample and were allowed to evaporate solvent overnight. An E-medium was prepared as per standard [24] and 20 ml was added in each flask. Another flask was supplemented with E- medium and plant growth promoters which served as negative control. Two fronds of Lemna minor plant were placed in these flasks which then kept in incubator for a week. Results were calculated in terms of % growth inhibition by counting numbers of fronds germinated in a week [25].

Conclusion

The present paper describes GC and GC-MS profile of the oily fraction recovered from the n- hexane soluble sub-fraction of C. intybus and subsequent determination of its phytotoxicity against Lemna minor.

Acknowledgement

Authors are thankful to Higher Education Commission, Pakistan for financial assistance and H.E.J Research Institute of Chemistry for spectral analysis and other technical support.

References

1. S. I. Ali, M. Qaiser, Flora of Pakistan, Missouri Botanical Press, St. Louis, Missouri, p.1 (2002).

2. R. Sharma, Medicinal plant of India-An Encyclopedia, Daya Publishing House Delhi, p. 57 (2003).

3. B. N. Sastari, The Wealth of India, Council of Scientific and Industrial Research, New Delhi, p. 555 (1992).

4. S. R. Baquar, Medicinal and Poisonous Plants of Pakistan, Printas Ltd. Karachi, p. 107 (1989).

5. N. D. Parajapati, S. S. Purohit, A. K. Sharma, T. Kumar, A Handbook Of Medicinal Plants, Agrobios (India) Jodhpur, p.139 (2003).

6. R. Norbaek, K. Nielsen and T. Kondo, Phytochemistry, 60, 357 (2002).

7. S. Shah, Z. Ali, A. Malik, I. A. Khan, S. Saied, Zeitschrift fur Naturforschung, 66 b, 729 (2011).

8. J. W. Timmermans, T. M. Slaghek, M. Iizuka, W. V. Ende, J. D. Roover, A. Laere, Journal of a. Carbohydrate Chemistry, 20, 375 (2001).

9. N. I. Khar'k, F. K. Khim, Chemistry of Natural Compounds, 9, 115 (1973).

10. V. G. Dem'yanenko, L. I. Dranik, Chemistry of Natural Compounds, 7, 104 (1971).

11. D. Thangadurai, S. Anitha, T. Pullaiah, P. N. Reddy and O. S. Ramachandraiah, Journal of Agricultural and Food Chemistry, 50, 3147 (2002).

12. S. Dwivedi, M. Khan, S. K. Srivastava, K. V. Syamasunnder and A. Srivastava, Flavour and Fragrance Journal, 19, 437 (2004).

13. J. Lesko, S. Holotik, J. Krupcik and V. Vesely, Journal of Chromatography, 119, 293 (1976).

14. C. T. Peng, R. L. Hua and D. Maltby, Journal of Chromatography, 589, 231 (1992).

15. V. K. Raina, S. C. Verma, S. Dhawan, M. Khan, S. Ramesh, S. C. Singh, A. Yadav and S. K. Srivastava, Flavour and Fragrance Journal, 21, 140 (2006).

16. K. M. Semnani, M. Azadbakht and A. Goodarzi, Flavour and Fragrance Journal, 19, 29 (2004).

17. J. A. Pino, R. Marbot and C. Vazquez, Journal of Essential Oil Research, 16, 318 (2004).

18. J. A. Pino, J. Mesa, Y. Munoz, M. P. Marti and R. Marbot, Journal of Agricultural and Food Chemistry, 53, 2213 (2005).

19. V. K. Raina, S. C. Verma, S. Dhawan, M. Khan, S. Ramesh, S. C. Singh and A. Yadav, Flavour and Fragrance Journal, 21, 140 (2006).

20. C. A. G. daCamara, S. L. K.Shepherd and D. R. G. Joaquim, Revista Brasileira de Farmacognosia, 12, 26 (2002).

21. I. Steinmetz, E. Schmolz and J. Ruther, Proceedings of the Royal Society London, 270, 385 (2003).

22. Atta-ur-Rehman, Studies in Natural Product Chemistry, Elsevier Science Publishers Netherlands, 9, 383 (1991).

23. R. Ahmed, A. Mahmood, F. Rashid, Z. Ahmed, M. Nadir, Z. Naseer and S. Kosar, Journal of Saudi Chemical Society, 11, 121 (2007).

24. A. Mahmood, R. Ahmed and S. Kosar, Journal of Saudi Chemical Society, 13, 273 (2009).

25. Y. Hideji, Oshida, A. Ikuta, H. Inatomi and T. Adachi, Phytochemistry, 21, 1935 (1982).

1Department of Chemistry, University of Karachi, Karachi-75720, Pakistan., 2P. N. E. C. National University of Sciences and Technology, Karachi-75270. Pakistan., 3HEJ Research institute of chemistry, International Center for Chemical and Biological Sciences,, University of Karachi, Karachi-75720, Pakistan., sumayyas@uok.edu.pk
COPYRIGHT 2012 Asianet-Pakistan
No portion of this article can be reproduced without the express written permission from the copyright holder.
Copyright 2012 Gale, Cengage Learning. All rights reserved.

Article Details
Printer friendly Cite/link Email Feedback
Author:Shah, Shazia; Mahmood, Azhar; Saied, Sumayya; Malik, Abdul
Publication:Journal of the Chemical Society of Pakistan
Article Type:Report
Geographic Code:9PAKI
Date:Dec 31, 2012
Words:2433
Previous Article:Effect of Ce-Co Substitution on the Structural, Electrical and Dielectric Properties of Mn Spinel Nanoferrites.
Next Article:Voltammetric Technique, A Panacea for Analytical Examination Of Environmental Samples.
Topics:

Terms of use | Privacy policy | Copyright © 2019 Farlex, Inc. | Feedback | For webmasters