Linear, Steroidal, and Triterpene Esters, and Steryl Glycosides from Festuca argentina
Adriana C. Casabuono and Alicia B. Pomilio*
PROPLAME-CONICET, Departamento de Química Orgánica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria, 1428 Buenos Aires, Argentina
ABSTRACT: Ester waxes and steryl glycosides of the grass Festuca argentina were studied. Saponiﬁcation of the waxes from the petroleum ether extract led to n-hexacosanol as the major single linear alcohol, along with pentacyclic triterpenols, such as βamyrin, germanicol, isobaurenol, lupeol, hopenol-a and hopeol, and low amounts of sterols, such as cholesterol, campesterol, stigmasterol, sitosterol and dihydrositosterol, identiﬁed by gas chromatography/mass spectrometry (GC/MS). Fatty acids were identiﬁed as methyl esters as C12:0, C14:0, C16:0, C18:0, C18:2, and C20:0. The occurrence of a wide chainlength range of fatty acids and a single linear alcohol closely matched for other reports on the tribe Festuceae. On the contrary, pentacyclic triterpenols with a variety of skeletons, especially isobauerenol, are not usual as esters of fatty acids in the Gramineae. Low amounts of steryl glycosides were also obtained from the methylene chloride percolate of the methanol extract. Upon acetylation followed by hydrolysis, aglycones were identified by capillary gas–liquid chromatography (GLC) and GC/MS. As ∆7-cholesterol, campesterol, stigmasterol, sitosterol, dihydrositosterol, and the sugars as glucose, xylose, and arabinose by GLC of the respective alditol acetates. This is the ﬁrst report on the linear, steryl, and triterpenyl esters of F. argentina. It is noteworthy that ∆7-steryl glycosides are rare, and steryl monoarabinosides have not been previously reported on the family Gramineae. Lipids 32, 205–210 (1997).
Festuca argentina (Speg.) Par. (Gramineae; common name: “coirón negro,” “coirón duro”) is a perennial xerophyte Patagonian grass popularly known as toxic to sheep (1) that grows in the pre-Andinian region, and eastward around the Gulf of San Jorge (2) in the Patagonian coast of Argentina. We have previously reported on free sterols and triterpenes from this species (3). The ∆5-, ∆7-, and dihydrosterols were identiﬁed as well as pentacyclic triterpenones with a variety of skeletons, such as oleanene/ursene (∆12-oleanen-3-one or βamyrenone, ∆18-oleanen-3-one = germanicone), lupene (lupen3-one), hopene (∆22(29)-hopen-3-one) and multiflorene (∆8*To whom correspondence should be addressed at PROPLAME-CONICET, Departamento de Química Orgánica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires. Pabellón 2, Ciudad Universitaria, 1428 Buenos Aires, Argentina. Abbreviations: GC/MS, gas chromatography/mass spectrometry; GLC, gas–liquid chromatography; MPLC, medium-pressure liquid chromatography; MS, mass spectra; MW, molecular weight; PE, petroleum ether; Rf, retention front; Rt, retention time; TLC, thin-layer chromatography.
multiﬂorenone = isomultiﬂorenone) and a cyclopropane tetracyclic triterpenone (cycloartanone), the two latter being rare in the Gramineae. However, the expected 4-methylsterols, 3methoxytriterpenes and arborene and/or fernene triterpenoids were not detected in this species, although widespread in the Gramineae (4). In continuation of our work on members of the tribe Festuceae, we want to report in this paper the composition of the ester waxes and steryl glycosides of F. argentina that, in contrast to other grasses, showed to be rich not only in a linear alcohol but also in both bound sterols and triterpenols. Wax esters from other members of the Gramineae usually accounted for esters of fatty acids–n-alcohols, e.g., Bromus inermis, F. ovina (5), Secale cereale (6), three wheat species (7–9), and Agropyron desertorum and A. intermedium (10), due to interesteriﬁcation of a wide range of acids with a much smaller chainlength range n-alcohols. Combined alcohols from waxes of festucoid grasses (5,11) generally contain one major component, C26 or C28, with the same chainlength as the major free alcohol of the wax. However, in Agropyron smithii (tribe Triticeae) a wide chainlength range of both acids and alcohols from the ester waxes were obtained without any prevalent long-chain major component (12), the alcohol portion also containing unusual appreciable percentage of α- and β-amyrins (40%). A wide range of chainlength linear alcohols and triterpenol esters was also typical of species of the subfamily Eragrostoideae (13). Detailed information on the identities of naturally-occurring phytosterols as glycosides is far from complete. So far, up to now these compounds have not been described as characteristic of any family, and they are scarcely distributed, except for sitosterol 3-O-glucoside, in agreement with earlier biosynthetic studies in Triticum aestivum, showing that UDP-glucose is the most active glycosyl donor (14). We have detected in F. argentina several steryl glycosides composed of a variety of steryl and sugar moieties. The occurrence of these compounds may be related to a sterol storage function, instead of simple waste products as considered by some plant physiologists especially interested in the role of free sterols (14). MATERIALS AND METHODS Plant material. Plants of F. argentina (Speg.) Par. (family: Gramineae) were collected by Prof. Ing. Angel Soriano in Río
Lipids, Vol. 32, no. 2 (1997)
Copyright © 1997 by AOCS Press
53 due to aldehydes. Medium-pressure liquid chromatography (MPLC) was carried out on silica gel H (Merck) columns with an impulse solvent bomb Prominent-Electronic 1001 SCJ (Heidelberg. it was composed of leaves (blades) and adventitious fibrous roots. Ac2O/pyridine. termed 16. CA) with flame-ionization detection (FID) and nitrogen as gas carrier.206
A.C. yielding sterols. CA). Elisa Nicora (Instituto de Botánica Darwinion. and 2c) was afforded by repeated preparative TLC (nhexane/EtOAc. Germany). 97:3). Extraction of the plant material. Patagonia). Hydrolysis of the steryl glycosides. The acetylated glycosides were hydrolyzed with 6% HCl in MeOH. J&W Scientific. and identified by Dr. Flash dry chromatography was performed under vacuum through a fritted Pyrex glass funnel membrane (Thomas Scientific. All solvent mixtures are expressed in volume (vol/vol). 95:5) contained steryl glycosides (3 mg. rendering 1. The reaction mixture was heated until dissolution of the glycosides and drops of water were added until there was slight turbidity. fresh material was air-dried in the laboratory in an aircirculating oven at 25oC to remove soil adhered to roots and blades. room temperature. roots. The reaction was monitored by TLC. H2O. Flash dry chromatography of PE extract (1. or paleas. The plant material used in this study was harvested in February (summer season in southern hemisphere) in the so-called fruit stage/period. 30 m length × 0. rate 15°C/min for glycosides aglycones. Swedesboro. POMILIO
Mayo (EE-INTA Río Mayo. TLC (n-hexane/EtOAc.80 and 0. then it was ground and immediately extracted in a Soxhlet with PE (60–80°C) followed by MeOH. lemmas. 2. temperature programmed from 240–300°C at 3°C/min for wax alcohols.3% relative to dry plant. 2b. silica gel G and silica gel H were purchased from Merck (Darmstadt.
. 0.38 due to terpenoids. Preparative TLC (20 cm × 20 cm. The aqueous layer containing the sugars was desalted with Amberlite MB-3/thymolphthalein (Sigma. This is a monocotyledonous matted grass with blades. and finally MeOH. NJ). high-performance thin-layer chromatography: EtOAc/MeOH. The CH2Cl2 layer was dried with anhydrous Na2SO4 and evaporated at 40°C. CASABUONO AND A. Freshly prepared plates were kept at room temperature for 24 h and further activated at 110–120°C for 1 h prior to use. Materials and chromatographic conditions. Argentina. 1000 µm layer) were prepared in our laboratory using a suspension of silica gel G (30 g) in distilled water (60 mL). Vol. and 200–300°C. glumes. Organic solvents were obtained from Sintorgan (Buenos Aires. 12 h. GLC was performed on a Hewlett-Packard 5840 A chromatograph (Palo Alto. Water was added to the residue and the aqueous layer was further extracted with CHCl3.6 % relative to dry weight) using gradients of n-hexane/EtOAc as previously reported (3) followed by TLC analysis led to six main fractions. Spikelets of grass species within a genus differ in the rachilla joints. Saponiﬁcation was performed by stirring the waxes (0. and a light-orange spot at Rf 0. as well as the sterols were conﬁrmed by capillary GLC with standards. 32. Capillary GC/MS were performed at 70 eV on a Hewlett-Packard 5890A chromatograph coupled with a selective mass detector 5970B. 0. and further hydrolyzed. voucher specimen were deposited in the Instituto de Botánica Darwinion (Argentina) SI 28004. 250 µm layer). The CH2Cl2 percolate was worked up as deLipids. After neutralization with NaHCO3 and evaporation of the solvent. the residue was partitioned with CH2Cl2/H2O (1:1). The MeOH extract was further percolated on polyamide with CH2Cl2.245 mm id. The remaining aqueous layer was acidified and the acidic compounds were extracted with CHCl3.8. Oven temperature was set at 180°C for 1 min with a 5°C/min temperature ramp up to a final temperature of 300°C for the alcohols from the ester waxes.25 mm. lupeol and hopeol. 98:2) of fraction 2 gave a main pink spot at Rf 0. βamyrin. Argentina). 0. CA). St. and 2. Folsom. and spikelets with minute structures which are important markers of taxa. Argentina) followed by distillation. polyamide for chromatography was purchased from Woelm (Eschwede. rate 10°C/min for the aglycones from the glycosylsterols. Germany). The identities of n-hexacosanol. The respective alcohol mixtures from the hydrolysis of the waxes and of the glycosylsterols were chromatographed on a bonded-phase fused silica capillary column (DB-1. These organic acids were methylated (CH2N2/Et2O) for 14 h at room temperature. Separation of these components (subfractions 2a. Province of Chubut. dried over anhydrous Na2SO4 and the chloroform was evaporated under vacuum to give the alcohols. a brown spot at Rf 0.005% relative to dry plant) which were acetylated (Ac2O/anhydrous pyridine) to overcome their insolubility in organic solvents (Rf 0. Bands were eluted with chloroform and monitored by analytical TLC.8). Analytical thin-layer chromatography (TLC) plates (silica gel G. MO) to prepare the alditol acetates (NaBH4. Germany) with a pulse damper. respectively. A Liebermann-Burchard positive (16) fraction that gave a diffuse pink spot of Rf 0. F-254.13 g) under reflux with 20% KOH in MeOH for 4 h. and devoid of spikelets. Saponiﬁcation of waxes.60 typical of waxes. and the waxes were characterized by infrared spectra (KBr) using a Perkin-Elmer 710-B spectrophotometer (Palo Alto.0.85. the mixtures were examined by GC/MS on a 25 m × 0. Louis. 1:0.B.33 µm ﬁlm thickness Ultra-1 capillary column. In addition it has a distinct festucoid-type embryo. This chloroform layer was dried over anhydrous Na2SO4 and then evaporated to dryness to give the organic acids. Then it was kept at 75°C in a sealed tube for 2 h. 2 (1997)
scribed before (15) yielding a subfraction 2 that was purified by MPLC (EtOAc).25 µm film thickness.65 by TLC (EtOAc/MeOH. The mixture was then evaporated to dryness. As soon as it was collected. except for petroleum ether (PE) and n-hexane which were freed from oleﬁnes in the usual way prior to distillation. 95:5). San Isidro. The organic layers were combined. Mass spectra (MS) data were processed by a Lab Base GC-MS data system (40 to 800 mass scanning). Gas–liquid chromatography (GLC) and gas chromatography/mass spectrometry (GC/MS) analysis of the compounds. Spots were visualized under ultraviolet (UV) light at 254 nm and/or by spraying the plates with H2SO4/AcOH (1:1) and heating at 110–120°C for 2 min. no. In addition. and 200–300°C. germanicol.
and an intense ion m/z 233 in agreement with dihydrositosterol (19).76. The MS of the respective sterols showed M+ 386. RESULTS AND DISCUSSION Ester waxes. 30 m length × 0.
16:0 38.. 1475 (δC-H). due to the fact that both skeletons are undistinguishable by MS. which accounted for 61% of the total alcohol wax composition. Identification was conﬁrmed by GC/MS spectrometry on a 1. The compound of Rt 20. see the Materials and Methods section.5%) typical of a ∆18-oleanene or ∆18-friedelene (20).2%) and low amounts of sterols (7. 1A). m/z 247 as base peak and other ions (Table 2) was characteristic (21) of a D:C-friedooleanane/D:C-friedo-ursane skeleton with one unsaturation either at ∆7-(multiflorenol/bauerenol) or at ∆8-(isomultiflorenol/isobauerenol). 412 and 414. and an intense ion at m/z 177 (98. prevailing the bauerene series (20) by the intense m/z 247 and the low abundant m/z 205. Supelco).g. along with another sterol of Rt and M+ higher than that of sitosterol. Sugars from steryl glycosides were analyzed by capillary GLC of their alditol acetates against synthetic standards (SP 2330. respectively. 1742 (νC=O). Upon hydrolysis. and M+ at m/z 214.99 min. The GC/MS of the pentacyclic triterpenoid alcohols were in the range of retention time (Rt) 20.2 m length × 2 mm i. the low abundance (14. a homologous series of saturated fatty acids with even carbon atom from C12 to C20 was obtained. M+ 426. 298. M+ and M+-Me of isobauerenol (22) and β-bauerenol (23). arising from hydroxyl-containing rings A/B of saturated triterpenols without directing groups. 1180 (νC-O) and 740 cm−1 (δC-H)]. Fig. at 190°C. The compound of Rt 20.ESTERS AND STERYL GLYCOSIDES FROM FESTUCA ARGENTINA
Fatty acids from the wax hydrolysis were separated as methyl esters on a 1. For details. along with the low intensity or absence of M+−
Lipids. along with triterpene alcohols (31.34 min. Repeated chromatographic purification techniques applied to the PE extract of F. base peak. corresponding to cholesterol. Gas chromatography/mass spectrometry of the alcohols from ester wax hydrolysis. PA) programmed from 60–270°C at 8°C/min. no. 242. argentina yielded a subfraction with an infrared spectrum typical of high molecular weight (MW) ester waxes [2895 (νC-H). further conﬁrmed by GLC comparison with a germanicol sample.0
1. Compounds with Rt 21.34 to 22.9 —
4.d. Vol. In addition. thus leading to isobauerenol (bauer-8-en-3-β-ol) only once reported in Gramineae (24). and m/z 218 as
TABLE 1 Composition of Acids and n-Alcohols from Festuca argentina Waxes 12:0 14:0 Esteriﬁed acidsa Esteriﬁed alcoholsb
FIG. n-hexacosanol (C26:0). argentina is shown in Table 1. 0. Identification of nhexacosanol is based on cochromatography with a known standard and GC/MS as indicated in Table 2.7 —
18:2 19. Bellefonte. Identification confirmed by gas chromatography/mass spectrometry (GC/MS) on a SP-2340 column. The compound of Rt 20.18. By contrast. and the fragmentation pattern typical of ∆5-sterols (Table 2) (18).93 min. 60-80 mesh. campesterol.8 —
base peak showed a fragmentation pattern (Table 2) of a ∆12ursene/∆12-oleanene triterpenol (20). confirmed as β-amyrin by capillary GLC with authentic samples.8 m × 2 mm 8% NPGS packed column on Chromosorb W-AW-DMCS. e. 2 (1997)
. the wax alcohols (Table 1. The fact that m/z 189 is more intense than m/z 191.0 —
Fatty acids analyzed as methyl esters by gas–liquid chromatography (GLC) on 8% NPGS and comparison with standards. (A) general chromatogram. (B) chromatogram of the ampliﬁed 15 to 24 min region. M+ 294 was shown to be a fatty acid of C18 containing two unsaturations (C18:2) by its mass fragmentation pattern.99 min showed similar MS (Table 2) concerning the molecular ion (M+ 426) and the highly abundant m/z 207. and 326. and 22. 400. which showed typical fragmentation pattern (17) by GC/MS analysis of their respective methyl esters with m/z 74 as base peak. M+ 426. 270. usually very high in ∆18-friedelene (20) provided evidence for a ∆18-oleanene. with hexacosanoic acid being the major component among the seven identiﬁed fatty acids.7 —
26:0 — 61. using authentic fatty acid methyl ester standards.7%) of ion m/z 203. b Alcohols were analyzed by capillary GLC on DB-1.52 min (Table 2) presented a M+ 426 followed by loss of 15 amu.6%) (Fig.25 mm id. 1. 1B). sitosterol. SP-2340 column (Supelco. 32. 22. The low levels of triterpenols and sterols were detected by GC/MS in the 15 to 24 min region of the ampliﬁed chromatogram (Fig. Comparison of relative abundances was required for the ∆7/∆8 location of the double bond. stigmasterol. 1) showed a single major linear alcohol.20 µm ﬁlm thickness.8 —
20:0 5. The acid composition of the ester waxes from F.
43. 218 (rings D and E + C − 11 + C − 12. 191 (26. 24. 315 (M+ − 85. 426 (M+. See Table 1 for abbreviation.8).76 min showed only two fragments in the high-mass region suggesting a difference in the double-bond location of the isopropenyl group relative to the two latter compounds. 301 (M+ − 85.9).1).1).2). 393 (M+ − Me − H2O.18
22. 190 (33. 427 (M+ + 1.9). 89.99
a Identiﬁcation conﬁrmed on a 25-m capillary column Ultra-1 by GC/MS (EI-70 eV). 71 (56.7).8).5). 14. 218 (16. 5.5). 107 (78. 98. 5. 273 (M+ − side chain. 29. 427 (M+ + 1. Consequently.9).9). 8.9).5). 14. 3.8). 393 (M+ − Me − H2O. 393 (M+ − Me − H2O. 207 (rings A and B. 7. 408 (M+ − H2O. 231 (M+ − side chain − 42. 189 (60.B.7).9). 271 (M+ − side chain − 2H. 382 (M+ − H2O.0). 401 (M+ + 1. Successive MPLC purification steps of the MeOH extract of F.3).4). 100.3). 7.9).9).1).0).7).3). 191 (35. 105 (46. 121 (65. 383 (M+ − isopropyl. respectively.7).5). 85.7).5). 205 (22. 8. 396 (M+ − H2O. 308 (0.5). 18. 0.9). Its chromatographic behavior (TLC.6). which usually lack a characteristic fragmentation pattern.0).9). 38.
Steryl glycosides.4). Vol.3).79
19.8). 10. thus supporting the occurrence of either lupane or hopane skeletons. 100.0).9). 29. 167 (4. 29. 353 (M+ − H2O − Me.p.8). 31. 385 (M+ − Me. 37.3). 111 (37. 100. 207 (79. 397 (M+ − Me.3). 153 (6. 300 (M+ − 112. 57 (100. 399 (M+ − Me. 17. 100.8). 41 (21. 415 (M+ + 1.0). 233 (M+ − side chain − 41 − H. 89. 189 (207 − H2O and/or rings D and E.7).0). 383 (M+ − H2O − Me.5).5). After acid hydrolysis of the acetate derivatives. 181 (3. 252 (0.4).9). rings D and E − H.52
20.8). 26.5). 121 (70.135 (64.9). 367 (M+ − H2O − Me.9). 37.5). 275 (M+ − 111. 195 (2. 15. 16. 427 (M+ + 1. 20.208
A. argentina gave rise to a LiebermannBurchard positive fraction (16) composed of steryl glycosides. 20. 386 (M+.
18. 247 (M+ − 139. 191 (32. 100. 207 (rings A and B.1). 231 (M+ − side chain − 42. 210 (1. 275 (M+ − 139.9).34
20. 5.6).9). 14. 426 (M+.0). 83 (83.0). 36. 39. 190 (43. 238 (1. 273 (M+ − side chain. 411 (M+ − Me of C − 17.3).5). 107 (76. 43 (76. 259 (rings A + B + C + Me − 26.C. 393 (M+ − Me − H2O.6).8). 2.5). 387 (M+ + 1.6).4). 381 (M+ − H2O − Me.2).4). 55 (62. 1. 125 (19. 205 (rings D and E or rings A and B.p. 417 (M+ + 1.2). 39.5). 97 (72. 27. 329 (M+ − 85. 17. 205 (rings D and E + C − 12. 49.5). 21. 217 (rings D and E + C − 11 + C − 12. 219 (18. 24. 400 (M+. 2. 207 (rings A and B.0). 19.2).5). 189 (207 − H2O and/or rings D and E − 29. 368 (M+ − H2O.9%). 73 Cholesterol 17. rings A and B.0). 261 (M+ − 139.6).6).93
21.6). 12. 266 (0. 2 (1997)
. 4. 204 (rings D and E + C − 12. 43.7).4%).8).0). CASABUONO AND A. 413 (M+ + 1. 0. 21.7). 206 (15. 19.3). 6. 105 (33. 178 (21.3).5).9).6).5). 190 (205 − Me. 1. 191 (18. 427 (M+ + 1. 109 (86.52 Characteristic mass peaks (m/z) (% relative abundance) 364 (M+ − H2O. thus leading to hopenol-a. 394 (M+ − H2O.68
19.9). 0. 8.0).6). 3.4). 416 (M+.3).7).5). 280 (0. 414 (M+. 100.99 min was confirmed by capillary GLC comparison with authentic samples of lupeol and hopeol (hopenol-b = hop-22(29)-en-3-β-ol).2). 273 (M+ − side chain.5). 15. 203 (37. 0.2).8).3).7). 48.9).6). 411 (M+ − Me. 32. 32.5).
43. 205 (rings D and E.4). 190 (7.2).2). C − 15 − C − 16.1). 43.7). 13. 18.10
18.7). 68. ). 59.8).9).18 and 22.76 22. The MS of the minor compound of Rt 22.8). 229 (57.005% relative to dry weight. 7. 218 (RDA. see the Materials and Methods section.59 (1. 412 (M+.9). Glucitol (16. 426 (M+. is indicative of an isopropenyl group in the molecule. 411 (M+ − Me.2).2).0). 189 (93. 204 (28. 19.4). no. 16.99
20. 289 (M+ − 111. 203 (22. which accounted for 0. 47.1). 100. 204 (24. and
Lipids.7). 177 (RDA in ring D + C − ring. 12. 28. 135 (60. 39. rings D and E. Rf region) was in agreement with nonacylated steryl monoglycosides. No changes were observed upon O-methylation (ethereal CH2N2). 401 (M+ − Me. 3. 38. both sterols (aglycones) and sugars (as alditol acetates) were identified by capillary GLC and capillary GC/MS analysis.1).9).3).1). the identity of compounds with Rt 21. 109 (70. 24.8).0).2). For details. 203 (218 − Me.7). 398 (M+ − H2O. b.8). 189 (RDA.7). 75. 139 (10. 426 (M+.0). 426 (M+. 303 (M+ − 111. 351 (369 − H2O. 369 (M+ − 43. 203 (218 − Me.2). 411 (M+ − Me. 208 (RDA. 2. 427 (M+ + 1. 426 (M+. 63. 427 (M+ + 1. 247 (RDA ring C and rupt.0). 218 (rings D and E. 379 (M+ − H2O − Me.0).3). 5.2).8). indicating the absence of uronic acids (25). xylitol (28. 215 (57.5). 224 (1. 69 (69. b. 234 (M+ − side chain − 41. 31.8).0). 108 (47. 11. 30. 371 (M+ − Me. POMILIO
TABLE 2 Alcohols from the Saponiﬁcation of Festuca argentina Waxes Alcohola n-Hexacosanol Rt (min) 12.6). 7. 336 (364 − CH2 = CH2. 204 (24. 23.
1610–1611. 23. VII. free (3). to Prof.. Syst. L. (1977) Composition of Epicuticular Waxes of Some Grasses. Allebone. 5. Phytochemistry 25. and only traces of esteriﬁed cholesterol. Phytochemistry 10. Budzikiewicz. Tulloch. J.3%). Phoebe.C. B.N. and Weenink.. Phytochemistry 23. Meksuriyen. 16. Tulloch. Agron. 3-O-monoxylosides. A.. 32. (1986) Gramineae. Wilson. 17. Conversely. (1983) Components of Bauhinia candicans. Ann.. Phytochemistry 24. 23. Parodi.. and Rangaswami. Chem.. (1976) Epicuticular Wax of Agropyron intermedium.. Free and Esterified Acids and Alcohols in Chenopodium album L. while ∆7-sterols are present only free (3) and/or as glycosyl derivatives. M. 13.
Thanks are due to CONICET (Argentina) and Universidad de Buenos Aires (Argentina) for financial support. 10. A.N.5%). Budzikiewicz. 9.O. Soc. such as stigmasterol 3-O-glucopyranoside (29). J. 85. Ayengar. Phytochemistry 13.B. INTA.8%) and esterified (6. L. and sitosterol 3-O-β-D-glucuronopyranoside (32).. 78–79.8%) occur in F.. A. stigmasterol (14. I. P..46%) forms. Soc. (1996) Festuca argentina and Festuca hieronymi: Chemical Relationships of Nonpolar Constituents. Argentina) for identification of the plant material. 7. 777–786. Ecol. C. Sengupta.P. Holden-Day. 26. glycosyl cholesterol (11. C. These ﬁndings closely resemble earlier reports on Sorghum vulgare (36). Tulloch.P. R. (1971) Leaf Wax of Durum Wheat. 360–361. A. Arg. Vol. Am. (1964) Interpretation of Mass Spectra of Organic Compounds. L. Nat... and Hoffman. Jr. 27. A. sitosterol 3-O-α-D-riburonofuranoside (25). A. (1982) Studies on the Constituents of
Lipids. 871–876.L. 1153–1156.) Vol. our results show that ∆5-sterols and dihydrositosterol are found in free (3).P.. campesterol (13. 6.3%). Moreover. Casabuono. Thus. (1988) Constituents of Rhamnus triquerta. 3. Nevertheless.P. argentina. and Williams. Djerassi. Tulloch. Inc. 25. and Pomilio. 479–483. Talapatra. Bot. and 3-O-monoarabinosides. and Shah. Fitoterapia 59. Buenos Aires. T.B. 3119–3126. steryl 3-Omonoarabinosides being so rare in nature that stigmasterol αL-arabinopyranoside has only been isolated once.P. (1970) Triterpenoids of the Gramineae. Pandey. (1964) Characterization of ∆4-3-Oxo-C21-steroids on Thin-Layer Chromatograms by in situ Colour Reactions. ed. N.22(E)-dien-3-α-ol (epichondrillasterol) (33). 273–282. Lolium perenne L. Iribarren. 136–151. sitosterol 3-O-β-D-xylopyranoside (30).
1. S. and Natori. from Wallenia yunquensis (35). 853–857. Plant Physiol.. and Hamilton. and Stellaria media L. Elucidation of the Course of the Characteristic Ring D Fragmentation of Unsaturated Steroids. C. ∆7-sitosterol (15.M. S.P.N. and Hoffman.3%) than the glycosylated (30. Ing. and to LANAIS-EMAR (Argentina) for GC/MS facilities. and Hoffman.. A. Am. S. 16. 99.P.. Can. A.. 55. (1977) Mass Spectrometry in Structural and Stereochemical Problems. (1981) Application of Mass Spectrometry in Structural Problems in Triterpenes. Sci. Knights.. 1685–1689. 20. (1969) Composition of the Leaf Wax of Little Club Wheat. 247–254.7%) were characterized and further confirmed by capillary GLC comparison with synthetic standards... D. Although sitosterol 3-O-glucopyranoside is widespread in nature (26–28). Phytochemistry 15. argentina. Chem. S. J. in Chemotaxonomie der Pflanzen. no. Casabuono.G. (1963) Mass Spectrometry in Structural and Stereochemical Problems. 47..A.L. Phytochemistry 20.ESTERS AND STERYL GLYCOSIDES FROM FESTUCA ARGENTINA
arabinitol (54. ∆7cholesterol (14. some of them as 3-Omonoglucosides.A. Basel. 8. 7686–7695. 1145–1151.J. 752–753. S. H.. Tetrahedron Lett. (1973) Leaf Wax of Triticum aestivum. L. C. 18. Dubcovsky for collection of the plant material. 15. 46. A. In summary. J.. J. Nicora. 3. Yeh. Phytochemistry 13.D. Partridge. Midgley. D. Tulloch. 5963–5968. 24. 11. and Pomilio.L. (1984) Epicuticular Waxes of Four Eragrostoid Grasses.M. J. esterified and glycosylated 4demethylsterols were found in F. whereas pentacyclic triterpenols merely occurred as esters of high-MW fatty acids. L. 22. 24. Hegnauer. (1974) Epicuticular Waxes of Secale cereale and Triticale hexaploide Leaves. 3688–3699. the relative percentage of the sterols in free and bound forms are distinctly variable. sitosterol. 29.. and Hoffman. and Djerassi. K. Ogunkoya.L.. Sunder. and Chiang. pp.A. esteryl and glycosyl forms.B. (1985) Sitosterol 3-O-α-DRiburonofuranoside from Bauhinia candicans. 14. M. Phytochemistry 35.H. Phytochemistry 15.D.6-dihydrositosterol (30. Vol.H. E. D-glucose is the usual sugar moiety (34) of monoglycosides of a variety of ∆5-phytosterols. 2. A. Ohmoto. A. XXXII.9%). Chem. R. (1986) Two Triterpenes from Davidsonia pruriens. Lisboa. 2535–2540. 26. Gas Chromatogr. 620–657. Nanayakkara.B.. J. E. (1994) Lignans and a Stilbene from Festuca argentina. G. R. Tulloch. J. Boston and Stuttgart. 249.P.L. free cholesterol (1. Rev. 21. only few other ∆5-steryl 3-O-monoglycosides have been reported. Nicora (Instituto de Botánica Darwinion. A. 209–236. (1978) Gramíneas. Prod. in Flora Patagónica (Correa. 2 (1997)
.. Rev. pp.H. A. and Cordell. H. Biochem. Phytochemistry 9.. Phytochemistry 12.. Can. To the best of our knowledge. 12.. and Pomilio. 2137–2148. L.B. (1968) A New Pentacyclic Triterpene Alcohol from Evodia fraxinifolia Hook F. Ram...3%) (3).. the isolation and identiﬁcation of monoglycosides of dihydrosterols and ∆7-sterols from natural sources is rare. C.E. Ikuse. and Talapatra. 163–229.. B. V. A. B. 121–126. (1950) Las Gramíneas Tóxicas para el Ganado en la República Argentina.P.
4.R. (1975) Plant Sterols. Accordingly. and Djerassi. A. 10–14. (1967) Identiﬁcation of Plant Sterols Using Combined GLC/MS. pp. 57. the proportion of free sitosterol and dihydrositosterol appears to be higher (56. A. sitosterol 3-O-α-Dxyluronofuranoside (31).N.2%). 1619–1623. 2217–2223. L.C. such as 3-O-β-Dglucopyranoside and 3-O-β-D-galactopyranoside of (24R)stigmast-7. 28. Colección Cientíﬁca del Instituto Nacional de Tecnología Agropecuaria.g.P. Food Agric.M.3%). Iribarren. 17. and Hoffman. Soriano and Dr. Birkhäuser Verlag. 118–119. A. (1976) Epicuticular Wax of Agropyron smithii Leaves. e. 5. 19. and 5. to Dr. and Pomilio.P. Chromatogr. L. Grunwald. A... H. J. (1974) Crystalline Chemical Components of Cheilanthes longissima. Tulloch. Pentacyclic Triterpenes. (1972) Cuticular Leaf Waxes III. San Francisco... R. Dwivedi.K. J. S. 3-O-monoglycosides of cholesterol (11.. Tulloch.
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