ISSN 10681620, Russian Journal of Bioorganic Chemistry, 2012, Vol. 38, No. 3, pp. 324–327. © Pleiades Publishing, Ltd., 2012.
Chemical Investigation of Iphigenia stellata Blatter by GCMS1 A. B. Sabalea and A. A. Maneb, 2 a
Department of Botany, Shivaji University Kolhapur 416004 b R.B. Madkholkar Mahavidyalaya, Chandgad 416509 Received May 24, 2011; in final form, June 14, 2011
Abstract—GCMS analysis of Iphigenia stellata Blatter in methanol extract revealed the presence of twenty chemical compounds in corm, nine in capsule wall, seven in leaves and six in seeds. Amongst the different phytochemicals identified 2furancarboxaldehyde5(hydroxymethyl) (38% in corms), glycerine (35.4% in seeds) and nhexadecanoic acid (31.5% in leaves, 28%) in capsule wall) were significant. Iphigenia stellata is known for the presence of colchicine. However, in the methanol extract it was not detected. Keywords: Iphigenia stellata, GCMSmethanol extract, glycerine, nhexadecanoic acid, 2furancarboxalde hyde5(hydroxymethyl) DOI: 10.1134/S1068162012030120 21
INTRODUCTION
Iphigenia stellata Blatter which belongs to the fam ily colchicaceae is a threatened medicinal herb found at discrete places in the Western Ghats of Maharashtra such as Panchgani, Pune, Sindhudurg, Gangan bavada, Panhala, Radhanagari, Kolhapur [1–3].The seed and corms of Iphigenia contain colchicine which has application in medicine and plant breeding [4–7]. The seeds contain glycerine which is useful in drugs, food, beverages and used as health supplement because it is useful in body temperature regulation [8, 9]. It is useful as feedstock for new types, biopoly mer, bioplastic film, biogas, in medicine it is used in eye drops in keratopathy, in pharmaceutical industries in tooth paste, skin creams, eye and ear drops, soaps it is an important content to maintain emollient effect [10–12]. All the parts of plant contain nhexadecanoic acid which possesses a variety of pharmaceutical prop erties such as larvicide, insecticides, pesticide as well as antibacterial potential [13, 14]. Tropolone alkaloids were reported in 15 genera of wurmbaeoideae and Iphigenia is one of them [15]. In the present work methanol extract of different plant parts of I. stellata are analysed on GCMS to reveal different bio chemicals.
Gas chromatographs of chemical constituents in methanol extract of seed, capsule wall, corm and leaves are shown in Fig. 1. Chemical structures of some bioactive components present in Iphigenia stellata are represented in Fig.2. Six compounds were identified in the methanol extract of seeds major constituent being glycerin (35.43%). Nine compounds were observed in capsule wall and highest composition was present in nhexade canoic acid (28%). Twenty compounds were seen in the corm and prevailing compound was 2furan car boxaldehyde, 5(hydroxymethyl), (38%) which is used for clinical treatment of sickle cell anemia [16]. Leaf extract revealed the presence of seven compounds in the methanol extract, with nhexadecanoic acid giving maximum % (31.52%). Some of the phytophar maceutical industries use Iphigenia stellata as a source of chemical constituent [17]. Lauric unhydride and nhexadecanoic acid were found common in all the plant parts analysed. Lauric unhydride is used in chemical industries as acylating agent to produce sucrose6laurate, in antitumor treatments, in cos metics and in pharmacy [18–20]. A few alkaloids from seeds of I. stellata have been recorded [21]. EXPERIMENTAL
RESULTS AND DISCUSSION The GCMS analysis of the methanol extract of seeds, capsule wall, corm and leaves of I. stellata is pre sented in Tables 1–4. 1 The article is published in the original. 2 Corresponding author: email:
[email protected].
Material. Plant material⎯Iphigenia stellata was collected in mature stage from table land of Panch gani in Satara district of Maharashtra and brought to the laboratory. Different plant parts such as seeds, capsule wall, corm and leaves were separated, oven dried at 60°C for 4 days, powdered and stored in air tight bottles.
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CHEMICAL INVESTIGATION OF Iphigenia stellata BLATTER BY GCMS
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Table 1. Components in the methanol extract of seeds of I. stellata No.
RT
Name of the compound
Molecular formula
Molecular weight
Composition, %
1 2 3 4 5 6
10.04 14.31 24.78 33.35 35.03 37.94
Glycerine 4HPyran4one, 2,3dihydro3,5dihydroxy6methyl Lauric anhydride 1,2Benzene dicarboxylic acid bis(2methylpropyl) ester nHexadecanoic acid 9Octadecenoic acid (E)
C3H8O3 C6H8O4 C24H46O3 C16H22O4 C16H32O2 C18H34O2
92 144 382 278 256 282
35.43 15.54 12.84 6.56 12.53 17.10
Note: RT, retention time.
Table 2. Components in the methanol extract of capsule wall I. stellata Molecular Molecular Composi formula weight tion, %
No.
RT
Name of the compound
1 2 3 4 5 6 7 8 9
12.21 16.64 17.18 19.75 20.74 21.98 23.79 23.98 28.92
2Furancarboxaldehyde, 5(hydroxymethyl) DAllose Lauric anhydride Tetradecanoic acid 1Naphtnalenol, 5,6,7, 8tetrahydro2,5dimethyl8(1methylethyl) nHexadecanoic acid 9,12Octadecadienoic acid (Z, Z) Octadecanoic acid 1,2Benzenedicarboxylic acid, bis(2ethylhexyl) ester, DOP
C6H6O3 C6H12O6 C24H46O3 C14H28O2 C15H22NO C16H32O2 C18H32O2 C18H36O2 C24H38O4
126 180 382 228 218 256 280 284 390
18.12 5.52 6.59 5.25 4.97 27.89 20.63 4.44 6.59
Table 3. Components in the methanol extract of corm I. stellata No.
RT
Name of the compound
Molecular formula
Molecular weight
Composi tion, %
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
6.14 7.75 9.55 10.41 10.82 13.28 13.80 16.06 16.90 17.20 19.76 20.90 21.24 21.55 22.00 2218 23.34 23.81 28.92 29.16
Furfural 2Furancarboxaldehyde, 5methyl 1,3,5Triazine2,4,6triamine Benzene, 1bromo2ethynyl 4HPyran4one, 2,3dihydro3,5dihydroxy6methyl 2Furan carboxaldehyde, 5hydroxylmethyl Benzene, 1bromo4iodo 1Methane sulfinyl4methylbenzene 1,6AnhydroβDglycopyranose (levoglucosan) Lauric anhydride Tetradecanoic acid Pentadecanoic acid Azoxybenzene Hexdecanoic acid nHexadecanoic acid Dibutylphthalate 10,13Octadecadienoic acid methyl ester 9,12Octadecadienoic acid (Z, Z) 1,2Benzenedicarboxylic acid diisocotylester 1,2Benzenedicarboxylic acid dicyclohexylester
C5H4O2 C6H6O2 C3H6N6 C8H2Br C6H8O4 C6H6O3 C6H4Br I C8H10OS C6H10O5 C24H46O3 C14H28O2 C15H30O2 C12H10N2O C17H34O2 C16H32O2 C16H22O4 C19H34O2 C18H32O2 C24H38O4 C20H26O4
96 110 126 180 144 126 282 154 162 382 228 242 198 270 256 278 294 280 390 330
1.83 0.89 1.16 3.61 5.50 37.98 1.55 2.84 7.20 7.20 1.75 0.49 0.86 0.96 10.14 2.07 2.68 2.15 2.15 2.38
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SABALE, MANE
Table 4. Components in the methanol extract of leaves of Iphigenia stellata Molecular Molecular Composi formula weight tion, %
No.
RT
Name of the compound
1 2 3 4 5 6 7
12.08 13.05 14.23 24.77 32.25 35.05 37.95
Acetophenone 1Butanol, 3methylacetate 4HPyran4one2,3dihydro3,5dihydroxy6methyl Lauric anhydride 1Naphthalenol, 5,6,7,8tetrahydro2,5 dimethyl8(1methylethyl) nHexadecanoic acid Octadec9enoic acid
C8H8O C7H14O2 C6H8O4 C24H46O3 C15H22O4 C16H32O2 C18H34O2
120 130 144 382 218 256 282
15.67 12.49 4.16 8.12 8.36 31.52 19.67
Note: Gas chromatographs of chemical constituents in methanol extract of seed, capsule wall, corm and leaves are shown in Fig. 1.
Plant sample extraction. Oven dried material (2 g) was defatted with petroleum ether and then extracted using soxhlet apparatus with methanol for six hours.
The extract was then filtered through Whatman filter paper no. 41 along with two gm of sodium sulphate to remove the sediment and trace of water. The filtrate
37.941
35.035
33.358
24.783
10.045
14.310
100.000 Iphigenia stellataSeed
TIC 1.00
28.927
23.798 23.984
21.981
TIC 1.00
28.925 29168
23.343 23.812
20
TIC 1.00
30
37.958
32.255
24.776
12.081 13.051 14.231
35.057
6 10 100.000 Iphigenia stellataLeaves
43 min
30 min
20.900 21.247 21.558 22.002 22.187
19.763
13.800
16.066 16.901 17.200
12.283 9.559 10.411 10.822
7.753
40
20
6 10 800.000 Iphigenia stellataCorm
6.143
30
20.746
19.754
12.210
16.648 17.186
20 10 200.000 Iphigenia stellataCapsule wall
TIC 1.00 10
20
30
40
43 min
Fig. 1. Gas chromatographs of chemical constituents in methanol extract of seed, capsule wall, corm and leaves of Iphigenia stel lata Blatter. RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY
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CHEMICAL INVESTIGATION OF Iphigenia stellata BLATTER BY GCMS (a)
(b)
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(c)
O
HO
OH OH
O OH
OH Fig. 2. Chemical structures of glycerine (a), nhexadecanoic acid (b), 2furancarboxaldehyde5(hydroxymethyl) (c).
was concentrated on a heating mantle to obtain four mL sample which was stored at 4°C. All the plant parts were used to prepare samples as described above. GCMS analysis. All the samples were passed through C18 column for clean up. Two µL sample was employed for GCMS analysis carried out on a GCMSMS, ITQ1100 (Thermoscientific) instru ment. The operating conditions were as follows: col umn–capillary columnDB130 mtrx 0.25 mm × 0.2 µm ID, injector. Programmable temperature vaporizer—PTV, carrier gas—99.99%, flow rate 1 mL/min, injector temperature was 250°C, oven temperature programme was 110°C/2 min—10°C/ min—200°C/0 min—5°C/min—280°C. Mass range was 40 to 550 m/z and multiplier voltage was 1300 mV. Identification of components. Interpretation of mass spectrum (GCMS) was conducted using data base of National Institute Standard and Technology (NIST). The spectrum of the unknown component was compared with the spectrum of known compo nents stored in the NIST library. The retention time, molecular mass, molecular formula, and composition percentage of the sample material was recorded. CONCLUSIONS The study revealed presence of several phytochem icals useful for pharmaceutical and other applications. These bioactive components present in I. stellata are economically important and with consistent demand in the field of medicine. Considering its significance this study will help in cultivation of I. stellata for com mercial extraction of chemical components. REFERENCES 1. Mukhopadhyay, M.J. and Mukhopadhay, S., Cytologia, 2008, vol. 73, no. 2, pp. 97–101. 2. Yadav, S.R. and Sardesai, M.M., Flora Kolhapur Distr. Shivaji Univ. Kolhapur, 2002, vol. 14, pp. 499–500. 3. Sharma, B.D., Karthikeyan, S., Singh, N.P., and Lax shminarasimhan P., Flora India Ser. 2, Flora Maha rashtra State—Monocotyledones, 1996, vol. 3, pp. 137– 140. RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY
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