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Sterol composition of the marine sponge Crambe crambe
Bkx~emicalSyslematicsand Ecology,Vol. 12, No. 3, pp. 323-324, 1984. Printed in Great Britain.
0305-1978/84 $3.00+0.00 Pergamon Press Ltd.
Sterol Composition of the Marine Sponge Crambe crambe F. CAFIERI, P. CIMINIELLO*, M. V. D'AURIA and C. SANTACROCE Istituto di Chimica Organica e Biologic,a, Universita di Napoli, 1-80134 Napoli, Italy; *lstituto di Chimica Biorganica, Universit~ di Napoli, 1-80138 Napoli, Italy
Key Word Index - Crambe crambe; Podfera; Poeciloscleridae; Mixillidae; marine sterols; chemotaxonomy. Abetract ~ The sterol content of the marine sponge Crambe crambe has been determined. The major components are cholest-7-en-3/3-ol, 24-methylcholesta-7,22-dien-3/J-ol and cholesta-7,22-dien-3/]-ol. Significative quantitites of the rare 4ornethyl-5~-cholest-8-en-3/~-ot are also present.
Introduction The sterol composition of the sponges has been widely studied and a number of unique sterols have been isolated from these organisms [1, 2] but little useful chemotaxonomic information can be drawn from these observations. For some taxa this can be explained by the small number of species examined; particularly so far only five sponges belonging to the order of Poeciloscleridae have been studied: Lissodendoryx noxiosa [4] and Tedania ignis [5] (Mixillidae), Neofibularia irata [3]
(Esperopsidae), Agelas oroides [5, 6] and A. mauritiana [7] (Agelasidae). Because of our interest in the chemistry of marine organisms we have now studied the sterol composition of the sponge Crambe crambe (Poeciloscleridae, Mixillidae). Results and Discussion The chemical structures and the percent composition of the sterols of C. crambe are reported in Table 1. The configuration of the asymmetric centres of the side chain cannot be determined
TABLE 1. STEROL COMPOSITION OF C. CRAMBE Skeletaltype R
R
H
H
.
~
1 (2.41%)
4 (10,08%)
2 (8.32%)
§ (22.26%)
3 (2.95%)
K
R
R
i"
9 (4,34%) 13 (6.65%)
6 (14.87%)
10 (7.60%)
7 (3.70%)
11 (1.58%)
8 (6.84%)
12 (1.97%)
(Received 8 August 1983) 323
14 (6.43%)
324
because epimeric sterols show virtually identical GLC, NMR and MS data. 4o-Methyl-5o-cholest-8-en-3/3-ol was identified by comparison of the physical and spectral properties of its acetyl derivative [o]o + 58~; m.p. 105-107°; MS: m/z 442 [M]*, 382 [M - AcOH]*, 329 [M -side chain]*, 269 [M -AcOH -side chain]*, 227 [M - A c O H - side chain-42]*; NMR: d 0.61 (s, H3-Cls), 0.86 (d, J = 6 Hz, H3-Ca), 0.99 (s, H3-Cls), 4.38 (m, H-C3) with those reported in the literature [8, 9]. The other sterols were identified by G C / M S and by comparison of retention data on GLC with those of authentic compounds. Remarkably C. crambe contains a predominance of &7 sterols, previously found in a few species of Porifera. Significative amounts of &5.7sterols, absent in the five sponges of order of Poeciloscleridae previously examined, are also present. Finally it is worthy of note that very recently a re-examination of the sterol fraction of the sponge A. oroides [6] showed that this organism contains, in addition to A7 sterols, substantial amounts of 14 and this could have some chemotaxonomic significance. Materials and Methods Fresh material (250 g) collected in autumn 1982 in the Bay of Naples was freeze-dried, ground finely and extracted with CHCI 3. The oily residue (2.41 g) was saponified with 10% KOH in aqueous 80% EtOH by refluxing for 2 h and the
F. CARERI, P. CIMINIELLO, M. V. D'AURIA AND C. SANTACROCE unsaponifiable matter (703 mg) was chromatographed on a silica gel column using as eluent C6Hs with increasing amounts of Et20. Fractions eluted with CsHs-Et20 (9:1 ) afforded 40 mg of crude 14, which was purified after acetylation [AczOpyridine (1:1) for 12 h at room temperature], by SiO2 PLC [aluant CsHs-Et20 (9:1 )]. Fractions eluted with CsHs-Et20 (8:2) gave 582 mg of a complex mixture of sterols which was further chromatographed on a SiO2/AgNO 3 column, using as eluant CsHs with increasing amounts of Et2O. Individual fractions were analysed by GLC (C. Erba Fractovap 4160 gas chromatograph on SE-52 capillary column at 250°), GC/MS (LKB 9000 S instrument using an SE-30 capillary column at 260°) and 1H NMR spectroscopy (Bruker WH 270 instrument, in CDCI 3, using TMS as internal reference). Quantitation was performed by GLC of sterols (cholestane as internal standard) using integrated areas of peaks.
References 1, Goad, L. J. (1978) Marine Natural Products (Scheuer, P. J., ed.) Vol. II, pp. 75-172. Academic Press, New York. 2. Djerasai, C. (1981) Pure App/. Chem. 53, 873. 3. Bergquist, P. R., Hofheinz, W. and Oesterelt, G. (1980) Biochem. Syst. Ecol. 8, 423. 4, Sheikh, Y. M. and Djerassi, C. (1974) Tetrahedron30, 4095. 5. De Rosa, M., Minale, L. and Sodano, G. (1973) Comp. 8iochem. Physiol. Part B 46, 823. 6. Di Giacomo, G., Dini, A., Falco, B., Marino, A. and Sica, D. (1984) Comp. 8iochem. Physiol. Part B (in press). 7. Bohlin, L., Henning, P., Gehrken, H. P., Scheurer, P. J. and Djerassi, C. (1980) Steroids35, 295. 8. Beastall, G. H., Tyndall, A. M., Reas, H. H. and Goodwin, T. W. (1974)Eur. J. Biochem. 41,301. 9. Kandutsch, A. A. and Russell, A. E. (1960) J. Biol. Chem. 235, 2253.
0305-1978/84 $3.00+0.00 Pergamon Press Ltd.
Sterol Composition of the Marine Sponge Crambe crambe F. CAFIERI, P. CIMINIELLO*, M. V. D'AURIA and C. SANTACROCE Istituto di Chimica Organica e Biologic,a, Universita di Napoli, 1-80134 Napoli, Italy; *lstituto di Chimica Biorganica, Universit~ di Napoli, 1-80138 Napoli, Italy
Key Word Index - Crambe crambe; Podfera; Poeciloscleridae; Mixillidae; marine sterols; chemotaxonomy. Abetract ~ The sterol content of the marine sponge Crambe crambe has been determined. The major components are cholest-7-en-3/3-ol, 24-methylcholesta-7,22-dien-3/J-ol and cholesta-7,22-dien-3/]-ol. Significative quantitites of the rare 4ornethyl-5~-cholest-8-en-3/~-ot are also present.
Introduction The sterol composition of the sponges has been widely studied and a number of unique sterols have been isolated from these organisms [1, 2] but little useful chemotaxonomic information can be drawn from these observations. For some taxa this can be explained by the small number of species examined; particularly so far only five sponges belonging to the order of Poeciloscleridae have been studied: Lissodendoryx noxiosa [4] and Tedania ignis [5] (Mixillidae), Neofibularia irata [3]
(Esperopsidae), Agelas oroides [5, 6] and A. mauritiana [7] (Agelasidae). Because of our interest in the chemistry of marine organisms we have now studied the sterol composition of the sponge Crambe crambe (Poeciloscleridae, Mixillidae). Results and Discussion The chemical structures and the percent composition of the sterols of C. crambe are reported in Table 1. The configuration of the asymmetric centres of the side chain cannot be determined
TABLE 1. STEROL COMPOSITION OF C. CRAMBE Skeletaltype R
R
H
H
.
~
1 (2.41%)
4 (10,08%)
2 (8.32%)
§ (22.26%)
3 (2.95%)
K
R
R
i"
9 (4,34%) 13 (6.65%)
6 (14.87%)
10 (7.60%)
7 (3.70%)
11 (1.58%)
8 (6.84%)
12 (1.97%)
(Received 8 August 1983) 323
14 (6.43%)
324
because epimeric sterols show virtually identical GLC, NMR and MS data. 4o-Methyl-5o-cholest-8-en-3/3-ol was identified by comparison of the physical and spectral properties of its acetyl derivative [o]o + 58~; m.p. 105-107°; MS: m/z 442 [M]*, 382 [M - AcOH]*, 329 [M -side chain]*, 269 [M -AcOH -side chain]*, 227 [M - A c O H - side chain-42]*; NMR: d 0.61 (s, H3-Cls), 0.86 (d, J = 6 Hz, H3-Ca), 0.99 (s, H3-Cls), 4.38 (m, H-C3) with those reported in the literature [8, 9]. The other sterols were identified by G C / M S and by comparison of retention data on GLC with those of authentic compounds. Remarkably C. crambe contains a predominance of &7 sterols, previously found in a few species of Porifera. Significative amounts of &5.7sterols, absent in the five sponges of order of Poeciloscleridae previously examined, are also present. Finally it is worthy of note that very recently a re-examination of the sterol fraction of the sponge A. oroides [6] showed that this organism contains, in addition to A7 sterols, substantial amounts of 14 and this could have some chemotaxonomic significance. Materials and Methods Fresh material (250 g) collected in autumn 1982 in the Bay of Naples was freeze-dried, ground finely and extracted with CHCI 3. The oily residue (2.41 g) was saponified with 10% KOH in aqueous 80% EtOH by refluxing for 2 h and the
F. CARERI, P. CIMINIELLO, M. V. D'AURIA AND C. SANTACROCE unsaponifiable matter (703 mg) was chromatographed on a silica gel column using as eluent C6Hs with increasing amounts of Et20. Fractions eluted with CsHs-Et20 (9:1 ) afforded 40 mg of crude 14, which was purified after acetylation [AczOpyridine (1:1) for 12 h at room temperature], by SiO2 PLC [aluant CsHs-Et20 (9:1 )]. Fractions eluted with CsHs-Et20 (8:2) gave 582 mg of a complex mixture of sterols which was further chromatographed on a SiO2/AgNO 3 column, using as eluant CsHs with increasing amounts of Et2O. Individual fractions were analysed by GLC (C. Erba Fractovap 4160 gas chromatograph on SE-52 capillary column at 250°), GC/MS (LKB 9000 S instrument using an SE-30 capillary column at 260°) and 1H NMR spectroscopy (Bruker WH 270 instrument, in CDCI 3, using TMS as internal reference). Quantitation was performed by GLC of sterols (cholestane as internal standard) using integrated areas of peaks.
References 1, Goad, L. J. (1978) Marine Natural Products (Scheuer, P. J., ed.) Vol. II, pp. 75-172. Academic Press, New York. 2. Djerasai, C. (1981) Pure App/. Chem. 53, 873. 3. Bergquist, P. R., Hofheinz, W. and Oesterelt, G. (1980) Biochem. Syst. Ecol. 8, 423. 4, Sheikh, Y. M. and Djerassi, C. (1974) Tetrahedron30, 4095. 5. De Rosa, M., Minale, L. and Sodano, G. (1973) Comp. 8iochem. Physiol. Part B 46, 823. 6. Di Giacomo, G., Dini, A., Falco, B., Marino, A. and Sica, D. (1984) Comp. 8iochem. Physiol. Part B (in press). 7. Bohlin, L., Henning, P., Gehrken, H. P., Scheurer, P. J. and Djerassi, C. (1980) Steroids35, 295. 8. Beastall, G. H., Tyndall, A. M., Reas, H. H. and Goodwin, T. W. (1974)Eur. J. Biochem. 41,301. 9. Kandutsch, A. A. and Russell, A. E. (1960) J. Biol. Chem. 235, 2253.