Faculty of Pharmacy, University of Istanbul, Istanbul, Turkey
By A. ULUBELEN and I. UYGUR
Petroleum ether and chloroform extracts of S a 1 v i a a e t h i o p i s yielded a hydrocarbon, two steroidal and two flavonoidal compounds. By using U V , IR and N M R spectral methods, analytical calculations as well as standard sample comparison, the compounds were found to be n-eicosan, (3-sitosterol, salvigenin, 5-hydroxy-3-'4,-'7-trimethoxyflavone and sitosteryl-30-D-glucoside. Salvia aethiopis (Labiatae) is a 30-50 cm tall perennial plant, wide spread in the European section of Turkey. T h e plant was collected in June 1972 at Babaeski (European Turkey), identified by Prof. Dr. A. BAYTOP(Istanbul). A voucher sample is deposited in the Herbarium of the Faculty of Pharmacy, University of Istanbul. There are quite a number of publications dealing with the flavonoids and triterpenic compounds of Salvia species. GELLAand PROKOSHEVA (1970) isolated diosmetin and luteolin from S.nutans. 7-Methyl luteolin, luteolin glycosides, and hispidulin and 6-methoxyluteolin were found in S.offrcinalis by BRIESKORN BIECHELE (1971). ULUBELEN, OZTURK and ISILDATICI (1968) obtained salvigenin from S.triloba. Ursolic and oleanolic acids were found in S.officinalis by BRIESK'ORN and POLONIUS (1962) by NICHOLAS(1962) by BRIESKORN and MELCHIOR and BRIES(1969). Micromeric acid was isolated from S.horminum by ULUBELEN KORN (1975). Anagadiol was obtained from S.broussuneti by GONZALES. ULUBELEN and AYANOGLU established the presence of a new triterpenic acid (virgatic acid) in S.virgata (1975). A literature survey revealed that there is no study done with S-aethiopis.
Experimental The following instruments were used for spectral analysis: Beckman DB for UV, Perkin-Elmer 577 for IR and Varian 60 D for NMR. Melting points were taken by a Reichert-Kofler model instrument without correction.
Extraction of the plant: About 2 k g of the air dried plant was extracted successively with petroleum ether, chloroform and alcohol, 70 g, 50 g and 80 g extracts were obtained respectively.
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Abstract
Compounds of Salvia aethiopis
319
Since the preliminary experiments indicated the same compounds the first two extracts were combined. Isolation of the compounds: 30 g of petroleum ether and chloroform extract was eluted on a silica gel (4x50 cm) column. The fractions were checked on TLC plates, similar ones were combined. Single compounds were identified as follows:
fl-sitostcrol: Chloroform fractions yielded about 1.5 g of a crude single compound, alcohol crystallization yielded a pure crystalline compound, m. p. 136O. Analysis for C,H,,O C, 84.05; H, 12.07 found C, 83.74; H, 11.97 Acetylation of 50 mg of the compound was carried out in 2 ml of pyridine using 2 ml of acetanhydride, m.p. 122-123O. C, 8157; H, 11.40 Analysis for C,,H,O, found C, 81.84; H, 11.67 TLC, m.rn.p. and IR spectra comparison with a standard sample proved that this was p-sitosterol. Salvigenin (5-hydroxy-6,7,4'-trirnethoxyflavone): Chloroform :alcohol fractions of the column yielded a yellow compound (0.750 g). Cellulose TLC plates showed the presence of two compounds, these were separated from each other by preparative TLC, using benzene : alcohol (9:l) as solvent system, Rt values 0.61 for salvigenin and 0.73 for methoxy luteolin. After separation, the above compound was crystallized from alcohol, needle like yellow crystals were obtained, m.p. 187-188'. Analysis for C,,H,,O, C, 65.85; H, 4.87 C, 65.69; H, 4.95 found UV max. 336 and 280 nm. The addition of AlCl, caused a bathochromic shift on UV spectrum, indicating a free 5-OH group. IR spectrum 3050 (bonded OH), 1650 (carbonyl group) and 825 cm-I (4'-OCHJ). TLC, mixed m.p. and IR spectra comparison with a standard sample of salvigenin, proved that these are identical compounds. Methoxyluteolin (5-hydroxy-3'4',7-trimethoxyflavone): After preparative TLC separation and alcohol crystallization, the second compound had a m.p. 164O. UV max 235 sh, 270, 324 and 340 nm. AIC1, caused a bathochrornic shift in UV spectrum, indicating a free 5-OH group. IR 1690, 1605, 1585, 1515, 1255 and 1055 cm-I. NMR (CDCI,, TMS) 3.92 (9H, s) (3xOCH,), 6.43, 6.62, 7.00, 7.30, 7.95 (aromatic protons), 11.60 p.p.m. (8) (intramolecular hydrogen bond). Standard sample comparison on TLC, m.m.p., IR and NMR spectra, it was found to be methoxyluteolinl. Methoxyluteolin was obtained and identified by A. Ulubelen and E. Ayanoglu from S. virgata.
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n-Eicosan: ~ h first d three fractions yielded 2.4 g of a single compound with some impurities, after a few crystallizations from CHCl, : EtOH (6:4) m.p. 35-36'. Analysis for CwHe C, 85.10; H, 14.89 found C, 84.80; H, 14.83 IR spectrum 2950,2850,14SO,1370,730 and 720 cm-I, showed that this was a saturated n-hydrocarbon. NMR spectrum gave methyl singlets at 0.9 and methylene peak at 1.25 ppm (6).Analysis, IR and NMR proved that this was n-eicosan.
320
Ulubelen and Uygur
Planta medica Vol. 29 1976
Sitosteryl-3f5-D-glucoside: Alcohol extract of the plant yielded about 1g of a white crystalline compound from the silica gel column when eluted with alcohol, m.p. 297O (degradation). C, 72.91; H, 10.41 Analysis for C,,H,,O, C, 72.09; H, 10.42 found IR spectrum 3450 (a broad peak for OH), 2950, 2880, 1480, 1375, 1110, 1080, 1060, 1030 cm-I (typical sugar C-0 peaks). TLC, m.m.p. and IR spectra comparison with the standard sample showed that this was sitosteryl-3fi-D-glucosideP.
BRIESKORN, C. H. and BIECHELE, W.: Arch. Pharm. 304,557 (1971) H. and MELCIOR,C.: ibid, 302,921 (1969) BRIESKORN,~. W.: Pharmazie, 17,705 (1962) BRIESKORN, C. H. and POLONIUS, L. I.: Khim. Prir. Soedin. 6,270 (1970); C.A. 73, 63196 w GELLA,E. C. and POROKOSHEVA, GONZALES, A. G., BRETON,J. L. and FRAGA,B. M.: chemical Communications 576 (1971) H. J.: Biol. Chem. 237,1476 (1962) NICHOLAS, E.: Accepted for publication in Phytochemistry ULUBELEN, A. and AYANOGLU, A. and BRIESKORN, C. H.: Phytochemistry 14,1459 (1975) ULUBELEN, S.: J. Pharm. Sci. 57,1037 (1968) ULUBELEN, A., O Z T ~ R K S., and ISILDATICI,
Address: Prof. Dr. A. Ulubelen, Faculty of Pharmacy, University of Istanbul, Turkey
Sitosteryl-3fi-D-glucoside was kindly provided by Prof. Dr. K. H. Pegel (Durban). 0
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References
By A. ULUBELEN and I. UYGUR
Petroleum ether and chloroform extracts of S a 1 v i a a e t h i o p i s yielded a hydrocarbon, two steroidal and two flavonoidal compounds. By using U V , IR and N M R spectral methods, analytical calculations as well as standard sample comparison, the compounds were found to be n-eicosan, (3-sitosterol, salvigenin, 5-hydroxy-3-'4,-'7-trimethoxyflavone and sitosteryl-30-D-glucoside. Salvia aethiopis (Labiatae) is a 30-50 cm tall perennial plant, wide spread in the European section of Turkey. T h e plant was collected in June 1972 at Babaeski (European Turkey), identified by Prof. Dr. A. BAYTOP(Istanbul). A voucher sample is deposited in the Herbarium of the Faculty of Pharmacy, University of Istanbul. There are quite a number of publications dealing with the flavonoids and triterpenic compounds of Salvia species. GELLAand PROKOSHEVA (1970) isolated diosmetin and luteolin from S.nutans. 7-Methyl luteolin, luteolin glycosides, and hispidulin and 6-methoxyluteolin were found in S.offrcinalis by BRIESKORN BIECHELE (1971). ULUBELEN, OZTURK and ISILDATICI (1968) obtained salvigenin from S.triloba. Ursolic and oleanolic acids were found in S.officinalis by BRIESK'ORN and POLONIUS (1962) by NICHOLAS(1962) by BRIESKORN and MELCHIOR and BRIES(1969). Micromeric acid was isolated from S.horminum by ULUBELEN KORN (1975). Anagadiol was obtained from S.broussuneti by GONZALES. ULUBELEN and AYANOGLU established the presence of a new triterpenic acid (virgatic acid) in S.virgata (1975). A literature survey revealed that there is no study done with S-aethiopis.
Experimental The following instruments were used for spectral analysis: Beckman DB for UV, Perkin-Elmer 577 for IR and Varian 60 D for NMR. Melting points were taken by a Reichert-Kofler model instrument without correction.
Extraction of the plant: About 2 k g of the air dried plant was extracted successively with petroleum ether, chloroform and alcohol, 70 g, 50 g and 80 g extracts were obtained respectively.
Downloaded by: Chinese University of Hong Kong. Copyrighted material.
Abstract
Compounds of Salvia aethiopis
319
Since the preliminary experiments indicated the same compounds the first two extracts were combined. Isolation of the compounds: 30 g of petroleum ether and chloroform extract was eluted on a silica gel (4x50 cm) column. The fractions were checked on TLC plates, similar ones were combined. Single compounds were identified as follows:
fl-sitostcrol: Chloroform fractions yielded about 1.5 g of a crude single compound, alcohol crystallization yielded a pure crystalline compound, m. p. 136O. Analysis for C,H,,O C, 84.05; H, 12.07 found C, 83.74; H, 11.97 Acetylation of 50 mg of the compound was carried out in 2 ml of pyridine using 2 ml of acetanhydride, m.p. 122-123O. C, 8157; H, 11.40 Analysis for C,,H,O, found C, 81.84; H, 11.67 TLC, m.rn.p. and IR spectra comparison with a standard sample proved that this was p-sitosterol. Salvigenin (5-hydroxy-6,7,4'-trirnethoxyflavone): Chloroform :alcohol fractions of the column yielded a yellow compound (0.750 g). Cellulose TLC plates showed the presence of two compounds, these were separated from each other by preparative TLC, using benzene : alcohol (9:l) as solvent system, Rt values 0.61 for salvigenin and 0.73 for methoxy luteolin. After separation, the above compound was crystallized from alcohol, needle like yellow crystals were obtained, m.p. 187-188'. Analysis for C,,H,,O, C, 65.85; H, 4.87 C, 65.69; H, 4.95 found UV max. 336 and 280 nm. The addition of AlCl, caused a bathochromic shift on UV spectrum, indicating a free 5-OH group. IR spectrum 3050 (bonded OH), 1650 (carbonyl group) and 825 cm-I (4'-OCHJ). TLC, mixed m.p. and IR spectra comparison with a standard sample of salvigenin, proved that these are identical compounds. Methoxyluteolin (5-hydroxy-3'4',7-trimethoxyflavone): After preparative TLC separation and alcohol crystallization, the second compound had a m.p. 164O. UV max 235 sh, 270, 324 and 340 nm. AIC1, caused a bathochrornic shift in UV spectrum, indicating a free 5-OH group. IR 1690, 1605, 1585, 1515, 1255 and 1055 cm-I. NMR (CDCI,, TMS) 3.92 (9H, s) (3xOCH,), 6.43, 6.62, 7.00, 7.30, 7.95 (aromatic protons), 11.60 p.p.m. (8) (intramolecular hydrogen bond). Standard sample comparison on TLC, m.m.p., IR and NMR spectra, it was found to be methoxyluteolinl. Methoxyluteolin was obtained and identified by A. Ulubelen and E. Ayanoglu from S. virgata.
Downloaded by: Chinese University of Hong Kong. Copyrighted material.
n-Eicosan: ~ h first d three fractions yielded 2.4 g of a single compound with some impurities, after a few crystallizations from CHCl, : EtOH (6:4) m.p. 35-36'. Analysis for CwHe C, 85.10; H, 14.89 found C, 84.80; H, 14.83 IR spectrum 2950,2850,14SO,1370,730 and 720 cm-I, showed that this was a saturated n-hydrocarbon. NMR spectrum gave methyl singlets at 0.9 and methylene peak at 1.25 ppm (6).Analysis, IR and NMR proved that this was n-eicosan.
320
Ulubelen and Uygur
Planta medica Vol. 29 1976
Sitosteryl-3f5-D-glucoside: Alcohol extract of the plant yielded about 1g of a white crystalline compound from the silica gel column when eluted with alcohol, m.p. 297O (degradation). C, 72.91; H, 10.41 Analysis for C,,H,,O, C, 72.09; H, 10.42 found IR spectrum 3450 (a broad peak for OH), 2950, 2880, 1480, 1375, 1110, 1080, 1060, 1030 cm-I (typical sugar C-0 peaks). TLC, m.m.p. and IR spectra comparison with the standard sample showed that this was sitosteryl-3fi-D-glucosideP.
BRIESKORN, C. H. and BIECHELE, W.: Arch. Pharm. 304,557 (1971) H. and MELCIOR,C.: ibid, 302,921 (1969) BRIESKORN,~. W.: Pharmazie, 17,705 (1962) BRIESKORN, C. H. and POLONIUS, L. I.: Khim. Prir. Soedin. 6,270 (1970); C.A. 73, 63196 w GELLA,E. C. and POROKOSHEVA, GONZALES, A. G., BRETON,J. L. and FRAGA,B. M.: chemical Communications 576 (1971) H. J.: Biol. Chem. 237,1476 (1962) NICHOLAS, E.: Accepted for publication in Phytochemistry ULUBELEN, A. and AYANOGLU, A. and BRIESKORN, C. H.: Phytochemistry 14,1459 (1975) ULUBELEN, S.: J. Pharm. Sci. 57,1037 (1968) ULUBELEN, A., O Z T ~ R K S., and ISILDATICI,
Address: Prof. Dr. A. Ulubelen, Faculty of Pharmacy, University of Istanbul, Turkey
Sitosteryl-3fi-D-glucoside was kindly provided by Prof. Dr. K. H. Pegel (Durban). 0
Downloaded by: Chinese University of Hong Kong. Copyrighted material.
References
