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Cyclopentanoid cyanohydrin glucosides from Passiflora mixta
Biochemical Systematics and Ecology 32 (2004) 695–698 www.elsevier.com/locate/biochemsyseco
Cyclopentanoid cyanohydrin glucosides from Passiflora mixta5 Trine Bylov, Henrik Franzyk, Jerzy W. Jaroszewski Department of Medicinal Chemistry, The Danish University of Pharmaceutical Sciences, Universitetsparken 2, DK-2100 Copenhagen, Denmark Received 28 June 2003; accepted 20 December 2003
Keywords: Passiflora mixta; Passifloraceae; Cyanogenic glucosides; Solid phase extraction; Charcoal
1. Subject and source Passiflora mixta L. f. (northern banana passion-fruit; syn. P. longiflora Lam., P. tomentosa Lam., P. tacso Cav., P. brachychlamys Harms), one of the Passiflora species (Passifloraceae) yielding edible fruits, is native to northwestern regions of South America. The plant was collected in the Botanical Garden, Aarhus. Voucher specimen (accession number DFHJJ34) was deposited in Herbarium C (Botanical Museum, University of Copenhagen, Copenhagen). 2. Previous work There are no literature reports of phytochemical investigations of P. mixta. 3. Present study Dried and milled plant material (aerial parts, 130 g) was extracted twice with boiling 80% aqueous methanol (800 ml). The combined extracts were evaporated, the residue dissolved in water (100 ml), and the solution washed with diethyl ether (3 200 ml). The aqueous phase was freeze-dried, the residue (22.3 g) was
5 Part 26 in the series ‘‘Natural Cyclopentanoid Cyanohydrin Glycosides’’. Corresponding author. Tel.: +45-3530-6372; fax: +45-3530-6040. E-mail address: [email protected] (J.W. Jaroszewski).
0305-1978/$ - see front matter # 2004 Elsevier Ltd. All rights reserved. doi:10.1016/j.bse.2003.12.007
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T. Bylov et al. / Biochemical Systematics and Ecology 32 (2004) 695–698
dissolved in water (350 ml), and the solution stirred for 30 min with charcoal (60 g). The mixture was filtered through a thin layer of charcoal on Celite, and the sorbent washed with water (2 500 ml). The glucosides were desorbed from the charcoal cake with ethanol (2 500 ml). The ethanolic solution was evaporated and the residue (1.9 g) was fractionated on a vacuum liquid chromatographic (VLC) column (7 7 cm, silica gel 60 H, Merck) (Taber, 1982; Zsoter et al., 1994) using a step gradient of methanol in dichloromethane. The fractions were monitored for cyanogenesis using the sandwich picrate assay (Brimer et al., 1983), TLC (silica gel, dichloromethane-methanol 10:1 and 4:1) and 1H-NMR (300 MHz, deuterium oxide), and pooled to give three major cyanogenic fractions. The most polar fraction (1.3 g) was further purified by VLC (6 6 cm silica gel column, step gradient of methanol in dichloromethane) to give 263 mg of a 1:1 mixture of 1 and sucrose (300 MHz 1H-NMR, deuterium oxide), from which pure 1 could be obtained by preparative HPLC (Phenomenex Luna 5 C18-2 column, 25 2 cm, 5 lm, 1% aqueous methanol, refractive index detection). Tetraphyllin B sulfate (1, total yield v v 0.1% dry weight), [a]D25 –40.7 (c 0.4, water), lit. –36 (Jaroszewski and Fog, 1989), 1 13 was identified by comparison of its H- and C-NMR spectra (600 MHz, methanol-d4) with those described in the literature (Jaroszewski and Fog, 1989). The second-most polar fraction (125 mg) was further purified by VLC (3 3 cm silica gel column, step gradient of methanol in dichloromethane) and preparative HPLC (same column as above, 10% aqueous methanol) to give 29 mg (0.02% dry weight) of a 1:1 mixture of tetraphyllin B (2) and volkenin (3), inseparable in this system (Jaroszewski et al., 1987a), and 9 mg (0.007% dry weight) of epivolkenin (4), apparently unaccompanied by its stereoisomer taraktophyllin (Jaroszewski et al., 1987b). The least polar fraction (88 mg) was purified by VLC (3 3 cm silica gel column, step gradient of methanol in dichloromethane) followed by preparative HPLC (same column as above, 25% aqueous methanol), to give 7 mg (0.005%) of a 3:1 mixture of tetraphyllin A (5) and deidaclin (6), inseparable in this system (Jaroszewski and Jensen, 1985). The glucosides 2–6 were identified using 1H- and 13 C-NMR spectra (600 and 100 MHz, respectively) of the free glucosides (in methanol-d4) and of their acetates (in chloroform-d) (Jaroszewski and Jensen, 1985; Jaroszewski et al., 1987a, 1987b). Although 2 and 3 as well as 5 and 6 were isolated as mixtures, unequivocal identification could be achieved by NMR spectroscopy before and after acetylation (Jaroszewski and Fog., 1989, 2002; Olafsdottir et al., 1989, 1990). The acetates were obtained by overnight treatment with acetic anhydride and pyridine (1:1).
4. Chemotaxonomic significance Cyclopentanoid cyanohydrin glucosides (cyanogenic glucosides) possess a distinct taxonomic value (Spencer et al., 1985; Olafsdottir et al., 1989, 1990; Jaroszewski et al., 2002; Clausen et al., 2002). All genera of Passifloraceae investigated thus far contain cyclopentanoid cyanohydrin glucosides (Jaroszewski et al., 2002; Clausen et al., 2002). Within Passiflora, the largest genus of Passifloraceae, the dis-
T. Bylov et al. / Biochemical Systematics and Ecology 32 (2004) 695–698
697
tribution of cyanohydrin glucosides shows distinct variations at the subgenus level (Jaroszewski et al., 2002).
P. mixta belongs to the subgenus Tacsonia, section Tacsonia, of Passiflora (Ulmer and Ulmer, 1997). While other subgenera of Passiflora, especially Decaloba and Passiflora, have been extensively investigated (Jaroszewski et al., 2002), P. mixta is the only representative of Tacsonia studied so far. The present study demonstrates that P. mixta produces cyanohydrins of type 1 and 2 (Jaroszewski et al., 2002). Thus, the present work contributes to the knowledge of the distribution of cyclopentanoids within the genus Passiflora. The sulfate 1 has previously been encountered only in a few species belonging to the subgenus Passiflora (Jaroszewski et al., 2002). In the present work, the initial separation of crude glucosides 1–6 from the extract was achieved by absorption on activated carbon (charcoal) and subsequent elution with ethanol. It was thus demonstrated that this method, previously used in isolation procedures for other types of plant glucosides (Weinges et al., 1986; Inouye, 1991), is applicable to a broad polarity range of cyclopentanoid cyanohydrin glucosides, including the very polar sulfate 1 as well as the relatively non-polar glucosides 5 and 6.
References Brimer, L., Christensen, S.B., Mølgaard, P., Nartey, F., 1983. J. Agric. Food Chem. 31, 789. Clausen, V., Frydenvang, K., Koopmann, R., Jørgensen, L.B., Abbiw, D.K., Ekpe, P., Jaroszewski, J.W., 2002. J. Nat. Prod. 65, 542.
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T. Bylov et al. / Biochemical Systematics and Ecology 32 (2004) 695–698
Inouye, H., 1991. Methods Plant Biochem. 7, 99. Jaroszewski, J.W., Jensen, B., 1985. Acta Chem. Scand. B 39, 867. Jaroszewski, J.W., Olafsdottir, E., Cornett, C., Schaumburg, K., 1987a. Acta Chem. Scand. B 41, 410. Jaroszewski, J.W., Andersen, J.V., Billeskov, I., 1987b. Tetrahedron 43, 2349. Jaroszewski, J.W., Fog, E., 1989. Phytochemistry 28, 1527. Jaroszewski, J.W., Olafsdottir, E.S., Wellendorph, P., Christensen, J., Franzyk, H., Somanadhan, B., Budnik, B., Jørgensen, L.B., Clausen, V., 2002. Phytochemistry 59, 501. Olafsdottir, E., Andersen, J.V., Jaroszewski, J.W., 1989. Phytochemistry 28, 127. Olafsdottir, E.S., Jaroszewski, J.W., Arbo, M.M., 1990. Biochem. Syst. Ecol. 18, 435. Spencer, K.C., Seigler, D.S., 1985. Biochem. Syst. Ecol. 13, 421. Spencer, K.C., Seigler, D.S., Fraley, S.W., 1985. Biochem. Syst. Ecol. 13, 433. Taber, D.F., 1982. J. Org. Chem. 47, 1351. Ulmer, B., Ulmer, T., 1997. Passionsblumen—Eine faszinierende Gattung. Laupenmu¨hlen Druck, Bochum. Weinges, K., Haremsa, S., Huber-Patz, U., Jahn, R., Rodewald, H., Irngartinger, H., Jaggy, H., Meltzer, E., 1986. Liebigs Ann. Chem. 46. Zsoter, Z., Eszenyi, T., Timar, T., 1994. J. Org. Chem. 59, 672.
Cyclopentanoid cyanohydrin glucosides from Passiflora mixta5 Trine Bylov, Henrik Franzyk, Jerzy W. Jaroszewski Department of Medicinal Chemistry, The Danish University of Pharmaceutical Sciences, Universitetsparken 2, DK-2100 Copenhagen, Denmark Received 28 June 2003; accepted 20 December 2003
Keywords: Passiflora mixta; Passifloraceae; Cyanogenic glucosides; Solid phase extraction; Charcoal
1. Subject and source Passiflora mixta L. f. (northern banana passion-fruit; syn. P. longiflora Lam., P. tomentosa Lam., P. tacso Cav., P. brachychlamys Harms), one of the Passiflora species (Passifloraceae) yielding edible fruits, is native to northwestern regions of South America. The plant was collected in the Botanical Garden, Aarhus. Voucher specimen (accession number DFHJJ34) was deposited in Herbarium C (Botanical Museum, University of Copenhagen, Copenhagen). 2. Previous work There are no literature reports of phytochemical investigations of P. mixta. 3. Present study Dried and milled plant material (aerial parts, 130 g) was extracted twice with boiling 80% aqueous methanol (800 ml). The combined extracts were evaporated, the residue dissolved in water (100 ml), and the solution washed with diethyl ether (3 200 ml). The aqueous phase was freeze-dried, the residue (22.3 g) was
5 Part 26 in the series ‘‘Natural Cyclopentanoid Cyanohydrin Glycosides’’. Corresponding author. Tel.: +45-3530-6372; fax: +45-3530-6040. E-mail address: [email protected] (J.W. Jaroszewski).
0305-1978/$ - see front matter # 2004 Elsevier Ltd. All rights reserved. doi:10.1016/j.bse.2003.12.007
696
T. Bylov et al. / Biochemical Systematics and Ecology 32 (2004) 695–698
dissolved in water (350 ml), and the solution stirred for 30 min with charcoal (60 g). The mixture was filtered through a thin layer of charcoal on Celite, and the sorbent washed with water (2 500 ml). The glucosides were desorbed from the charcoal cake with ethanol (2 500 ml). The ethanolic solution was evaporated and the residue (1.9 g) was fractionated on a vacuum liquid chromatographic (VLC) column (7 7 cm, silica gel 60 H, Merck) (Taber, 1982; Zsoter et al., 1994) using a step gradient of methanol in dichloromethane. The fractions were monitored for cyanogenesis using the sandwich picrate assay (Brimer et al., 1983), TLC (silica gel, dichloromethane-methanol 10:1 and 4:1) and 1H-NMR (300 MHz, deuterium oxide), and pooled to give three major cyanogenic fractions. The most polar fraction (1.3 g) was further purified by VLC (6 6 cm silica gel column, step gradient of methanol in dichloromethane) to give 263 mg of a 1:1 mixture of 1 and sucrose (300 MHz 1H-NMR, deuterium oxide), from which pure 1 could be obtained by preparative HPLC (Phenomenex Luna 5 C18-2 column, 25 2 cm, 5 lm, 1% aqueous methanol, refractive index detection). Tetraphyllin B sulfate (1, total yield v v 0.1% dry weight), [a]D25 –40.7 (c 0.4, water), lit. –36 (Jaroszewski and Fog, 1989), 1 13 was identified by comparison of its H- and C-NMR spectra (600 MHz, methanol-d4) with those described in the literature (Jaroszewski and Fog, 1989). The second-most polar fraction (125 mg) was further purified by VLC (3 3 cm silica gel column, step gradient of methanol in dichloromethane) and preparative HPLC (same column as above, 10% aqueous methanol) to give 29 mg (0.02% dry weight) of a 1:1 mixture of tetraphyllin B (2) and volkenin (3), inseparable in this system (Jaroszewski et al., 1987a), and 9 mg (0.007% dry weight) of epivolkenin (4), apparently unaccompanied by its stereoisomer taraktophyllin (Jaroszewski et al., 1987b). The least polar fraction (88 mg) was purified by VLC (3 3 cm silica gel column, step gradient of methanol in dichloromethane) followed by preparative HPLC (same column as above, 25% aqueous methanol), to give 7 mg (0.005%) of a 3:1 mixture of tetraphyllin A (5) and deidaclin (6), inseparable in this system (Jaroszewski and Jensen, 1985). The glucosides 2–6 were identified using 1H- and 13 C-NMR spectra (600 and 100 MHz, respectively) of the free glucosides (in methanol-d4) and of their acetates (in chloroform-d) (Jaroszewski and Jensen, 1985; Jaroszewski et al., 1987a, 1987b). Although 2 and 3 as well as 5 and 6 were isolated as mixtures, unequivocal identification could be achieved by NMR spectroscopy before and after acetylation (Jaroszewski and Fog., 1989, 2002; Olafsdottir et al., 1989, 1990). The acetates were obtained by overnight treatment with acetic anhydride and pyridine (1:1).
4. Chemotaxonomic significance Cyclopentanoid cyanohydrin glucosides (cyanogenic glucosides) possess a distinct taxonomic value (Spencer et al., 1985; Olafsdottir et al., 1989, 1990; Jaroszewski et al., 2002; Clausen et al., 2002). All genera of Passifloraceae investigated thus far contain cyclopentanoid cyanohydrin glucosides (Jaroszewski et al., 2002; Clausen et al., 2002). Within Passiflora, the largest genus of Passifloraceae, the dis-
T. Bylov et al. / Biochemical Systematics and Ecology 32 (2004) 695–698
697
tribution of cyanohydrin glucosides shows distinct variations at the subgenus level (Jaroszewski et al., 2002).
P. mixta belongs to the subgenus Tacsonia, section Tacsonia, of Passiflora (Ulmer and Ulmer, 1997). While other subgenera of Passiflora, especially Decaloba and Passiflora, have been extensively investigated (Jaroszewski et al., 2002), P. mixta is the only representative of Tacsonia studied so far. The present study demonstrates that P. mixta produces cyanohydrins of type 1 and 2 (Jaroszewski et al., 2002). Thus, the present work contributes to the knowledge of the distribution of cyclopentanoids within the genus Passiflora. The sulfate 1 has previously been encountered only in a few species belonging to the subgenus Passiflora (Jaroszewski et al., 2002). In the present work, the initial separation of crude glucosides 1–6 from the extract was achieved by absorption on activated carbon (charcoal) and subsequent elution with ethanol. It was thus demonstrated that this method, previously used in isolation procedures for other types of plant glucosides (Weinges et al., 1986; Inouye, 1991), is applicable to a broad polarity range of cyclopentanoid cyanohydrin glucosides, including the very polar sulfate 1 as well as the relatively non-polar glucosides 5 and 6.
References Brimer, L., Christensen, S.B., Mølgaard, P., Nartey, F., 1983. J. Agric. Food Chem. 31, 789. Clausen, V., Frydenvang, K., Koopmann, R., Jørgensen, L.B., Abbiw, D.K., Ekpe, P., Jaroszewski, J.W., 2002. J. Nat. Prod. 65, 542.
698
T. Bylov et al. / Biochemical Systematics and Ecology 32 (2004) 695–698
Inouye, H., 1991. Methods Plant Biochem. 7, 99. Jaroszewski, J.W., Jensen, B., 1985. Acta Chem. Scand. B 39, 867. Jaroszewski, J.W., Olafsdottir, E., Cornett, C., Schaumburg, K., 1987a. Acta Chem. Scand. B 41, 410. Jaroszewski, J.W., Andersen, J.V., Billeskov, I., 1987b. Tetrahedron 43, 2349. Jaroszewski, J.W., Fog, E., 1989. Phytochemistry 28, 1527. Jaroszewski, J.W., Olafsdottir, E.S., Wellendorph, P., Christensen, J., Franzyk, H., Somanadhan, B., Budnik, B., Jørgensen, L.B., Clausen, V., 2002. Phytochemistry 59, 501. Olafsdottir, E., Andersen, J.V., Jaroszewski, J.W., 1989. Phytochemistry 28, 127. Olafsdottir, E.S., Jaroszewski, J.W., Arbo, M.M., 1990. Biochem. Syst. Ecol. 18, 435. Spencer, K.C., Seigler, D.S., 1985. Biochem. Syst. Ecol. 13, 421. Spencer, K.C., Seigler, D.S., Fraley, S.W., 1985. Biochem. Syst. Ecol. 13, 433. Taber, D.F., 1982. J. Org. Chem. 47, 1351. Ulmer, B., Ulmer, T., 1997. Passionsblumen—Eine faszinierende Gattung. Laupenmu¨hlen Druck, Bochum. Weinges, K., Haremsa, S., Huber-Patz, U., Jahn, R., Rodewald, H., Irngartinger, H., Jaggy, H., Meltzer, E., 1986. Liebigs Ann. Chem. 46. Zsoter, Z., Eszenyi, T., Timar, T., 1994. J. Org. Chem. 59, 672.