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Flavonol glycosides from Tagetes elliptica
Phycochemistry, Vol. 31, No. printed in Great Britain.
12, pp. 43874388,
0031 9422/92 $5.00+0.00
1992
Pergamon PressLtd
FLAVONOL GLYCOSIDES FROM TAGEZ’ES ELLIPZTCA MARIOD'AGOSTINO,FRANCESCO
~~~irnento
DE SIMONE,ZHONG
LIANG ZHOU* and COSIMO PIZZA
di Chiiica delle Sostanze Naturali, Universiti di Napoli “Federico II”, Via Domenico Montesano, 49,80131 Napoli, Italy; *Shanghai Institute of Pharmaceutical Industry, Shanghai, People’s Republic of China (Received in revised form 16 April 1992)
Key Word Index-Tag&s elliptica; Asteraceae; quercetin 3-(3”,6“-diacetylgalactoside) and 3-(2”,3“,4”-triacetylgalactoside); flavonol glycosides.
Abstract-Two
new flavonol glycosides, quercetin 3-(~,~~~tylg~a~toside) and quercetin 3-(2”,3”,4”triacetylgalactoside) have been isolated from a methanolic extract of Tagetes elliptica and identified on the basis of chemical and spectral data. The known compounds quercetin 3-galactoside, isorh~e~ 3~rhamnosyl(l-*6)glucoside, myricetin 3&ucoside, querceti isorhamnetin 3-galactoside, quercetin 3-(6”-galloylgalactoside), quercetin 3rhamnoside, quercetin 3-rhamnosyl(l+Ci)galactoside and rhamnetin were identified from the same extract.
INTRODUCTION Tugetes eEl@ica Sm. is a herbaceous plant widespread in the Peruvian Sierra where it is used in folk medicine [i, 21. In an investigation of a methanoiic whole plant extract of T. elliptica two new flavonol glycosides (1 and 2) were isolated together with nine known compounds (3-11). The present paper describes the identification of 1 and 2. BESULTsANDDLSCUfSION
The ‘H NMR spectrum of l-3 showed the characteristic aglycone pattern of quercetin derivatives, namely a 2H AX and a 3H ABX system (see Experimental). The 13CNMR spectrum of these products exhibited all the upfield and downfield shiRs of the corresponding signals of quercetin consistent with 3-O-glycosidation [3]. The identity of quercetin as aglycone was further confirmed by W spectral data (see Experimental). The sugar moiety of 3 [3] was identified as B-Dgalactopyranose by FAB mass spectrometry, anomeric proton J value (7.5 Hz) and 13CNMR data [4] (see Table 1). However, the FAB mass spectrum of 1 showed singlets (3H) at 61.85 and 2.18 due to acetyl groups and diagnostic downfield shifts concerning H-3” (64.83, dd, J =9 and 4.2 Hz) and H-6”a (64.20, dd, J= 12 and 8.3 Hz). The corresponding signals for 3 [5] and anomeric coupling constant (J = 7.5 Hz) indicated that here the sugar moiety was 3,6-dia~tyl-~-D-g~~tose. This was confirmed by downfield shifts observed for the peaks of C-3“ (+ 2.3 ppm) and C-6” (+ 2.2 ppm) and by upfield shifts noted for the peaks of C-2” (- 2.8 ppm), C-4” (- 2.3 ppm) and C-S” ( - 4.0 ppm) in comparison with those of homologous carbons of 3 (see Table 1). The FAB mass spectrum and I-I-1” J value (7.5 Hz), along with the singlets (3H) at 6 1.86,2.05 and 2.16 showed that sugar residue of 2 was a ~-~t~a~tyl~iactose. The ‘HNMR spectrum indicated C-2”, C-3” and C-4” as acetylation sites, which was deduced by downfield shifts of H-4”, H-3” and H-2” signals in comparison with the co~es~nd~g signals for
3 (see Experimental) [S]. The identity of peaks was estab~shed on the basis of the multipIi~ty and decoupling experiments: irradiation at 85.01 (H-3”, dd, J =9 and 4.2 Hz) and 4.90 (H-2”, submerged by HOD signal) caused the simplification of signals centred, respectively, at 65.32 (H-4”, d, J=4.2 Hz) and 5.29 (H-l”, d, J = 7.5 Hz). The 2,3,4-triacetylation of a-D-g&iCtOSe was also consistent with i3CNMR data (see Table 1). The upfield shifts of C-l” (- 0.7 ppm) and C-5” (- 2.5 ppm) signais in comparison with those of the homologous carbons of 3 were ascribable, respectively, to acetyl groups linked at C-2” and C-4”. The total upfield shift (- 2.9 ppmf observed for C-3” and the chemical shifts of C-2” (670.6) and C-4” (668.9) were imputable to the combination of the acetylation downfield shift and upfield shift (y effect). Finally, 1 and 2 were both deacyiated to give 3, further confirming that gaiactose was the sugar moiety in these compounds. On the basis of these data, 1-3 were identified, respectively, as quercetin 3-O-~-D-(~‘,~-dia~tylgaI~topyranoside), quercetin 3-~-~-D-(2”,~,~-t~a~ty~g~actopyranoside) and the known quercetin 3-U+-galactopyranoside. Finally, the identification of known compounds isorhamnetin 3-rhamnosyl-( l-+6) glucoside (4), myricetin 3-glucoside (!5),quercetin (6), isorhamnetin 3-galactoside (7), quercetin 3~~-~lloylgal~toside (8), quercetin 3rhamnoside (9), quercetin 3-rhamnosyl-( 1-+6)galactoside (10) and rhamnetin (11) was based on comparison with literature data [3, 6-Q
EXPERIMENTAL FAB MS was carried out at 2-6 kV with a glycerolthioglycerol matrix in negative ion mode. The NMR spectra were carried out in CD,OD solutions at 250 MHz (IHNMR) and 62.5 MHz (‘“C NMR) with solvent as int. standard. Plant material. Tagetes elliptica was collected in Peru (province of Huacabamba, department of Piura) in October 1988. A voucher specimen is deposited in the Herbarium of Cattedra di
4387
Short Reports Table 1. “C NMR spectra of compounds CD,UD
Aglycone
c
1
2
3
2
158.5 135.7 179.5 163.0 loo.1 166.1 94.9 158.5 104.9 123.4 117.7 145.8 150.0 116.1 123.5 105.6 70.4 77.4 67,7 73.2 64*1 172.6 20.3 20.9
158.3 135.4 179.2 162.8 100.0 165.9 94.9 159.6 105.5 123.0 317.6 145.9 149.7 115.8 123.5 104.7 70.4 72.2 68.9 74.7 62.7 172.1 20.4 20.5 20.7
158.3 135.8 179.4 f 63.0 99,8 166.0 94.7 158.8 104,2 123.2 117.8 145.3 149.9 116.1 122.9 105.4 73.2 751 70.0 77.2 61.9
4 5 6
8 9 10
Galactow
C=O (Acetyl) Me (Acetyl)
l-3 in
(6)
2 3’ 4 5’ 6 1” 2” 3” 4” 5,I 6”
Botanica Farmaceutica (Dipartimento di Chimica delle Sostanze Naturali, Universitii “Fed&co IT”, Napoli). Extraction and isohtiun. The air-dried plant (400 g), defatted with petrol and CHCI,, was extracted with M&H at room temp. The crude extract (54 g), dispersed in H,U and reextracted with &OH, gave 16 g of a flavonoid mixture which was chromatographed on a Sephadex LH-20 column (80 x 4 cm) in 2-g batches. Fractions of 10 ml were eluted with MeQH, analysed by TLC (silica gel) in BuOH--HOAc-Hz0 (12: 3 : 5)and further fractionated by HPLC on a CIs p-Bondapak column (30 cm x 8 mm i.d., flow rate: 2.5 ml min- ’ ). Using MeOH-H&I I : 1as eluent, frs 9-15 (I.8 g) gave 4 f32 mg, R, 9 minb 10(66 mg R, 12 min), 1 (18 mg, R, 27 min) and 2 (24 mg, R, 41 min), whiL frs 33-40 (0.2 g) gave 6 (10 mg, R, 28 minf. With MeOH-H,O 2:3 as eluent, frs 16-22 (0.8 g) gave 3 (35 mg R, 30 min) and 9 (15 mg, R, 50 mm); frs 23-27 (0.2 g) gave 5 (14 mg, R, 19 min), 7 (10 mg, R, 29 min) and 11 (12 mg, R, 51 mm); frs 28-32 (0.3 g) gave 8 (34 mg, R, 19 mm). Alkaline /ry&oIysis. Compounds 1 and 2 (each 10 mg) in 0.5 M aq. RUH (1 ml) were heated at 110” in ;i stoppered reaction vial for 2 hr. The reaction mixture, adjusted to pH 7, was extracted with B&H. The organic phase was evapd to dryness, dissoived in CD,OD, and analysed by ‘H and t 3C NMR.
Quercetin 3-0-~-6-x)-(3”,5”-$i~~~~~~~~~~~~~~~~e~ (1). UV A,, nm: MeOH 256,302& 359; + AICI, 273,30&h, 436; + NaOAc 271, 317,413; NaOAc+H,BO, 258,293sh, 382; NaUMe 272, 321, 415. ‘HNMR: 67.80 (IH, i$ J=2 Hz, H-2’), 7.62 flH, dd, J=2 and 8.4 Hz, H-6’), 6.88 (lH, d, J =8.4 Hz, H-S), 6.41 (lH, d, J =2 Hz, H-B), 6.21 (lH, d, J=2 Hz, H-6), 5.22 (lH, d, Ja7.5 Hz, H-l”), 4.83 (lH, dd, J =9 and 4.2 Hz, H-3”), 4.20 (lH, dd, J = 12 and 8.3 Hz, 6”a), overlapping sign& in the 6 3.95-4.10 region (Hwb, H-2” and H-4”), 3.83 ffH, M, H-5”), 2.18 (3H, s, AC), 1.85 (3H, s, Ac). FAB MS (negative ion mode): m/z 547 fM -HI -, 504 [M-H-433-, 300 [M-H-247j-. Quercetin 3-o-B-o-(2”,3”,4”-triacetylgalactoside) (2). UV &laXnm: MeOH 257, 302sh, 358; +AlCl, 274, 30&h, 438; + NaOAc 272, 317, 414; +NaOAc+ H,BO, 258, 293sh 383; +NaOMe2?2,323,415. IHNMR: S?.65fIH,d,J=2 H&H-T), 7.50 (iH, dd, J =2 and 8.4 Hz, H-St), 6.87 (IH, d, J=8.4 Hz, HS), 6.35 flH, d, 5=2 Hz, H-81, 6.16 (IH, d, J=2 Hz, H-6), 5.32 (TN, Jz4.2 Hz, H-4”): 5.29 {lH, d, J=7.5 Hz, H-i”), 5.01 flH, dd, J=9 and 4.2 Hz, H-3”), 4.90 (H-2”, obscured by HOD), 3.90-4.10 region (H-5”, H-6”% H-6”b, overlapping signals), 2.16 (3H, s, Ac), 2.05 (3H, s, AC), 1.86 (3H, s, AC).FAR MS (negative ion mode): m/z 589 [M-H]-, 546 EM-H-43]-, 300 [M-H -239]-. Querceiirt ?-@&D-gnfuctoside (3). UV &,_ nm: M&H, 256* 301sh, 357; -I+AlCl, 274, 3OOsh,432; +NaOAc 273, 317, 393; NaOAc+H,BO, 254, 290, 377; NaOMe 273, 325, 412. ‘HNMR: 67.87 (lH, d, J=2 Hz, H-2’), 7.61 (lH, dd, J=2 and 8.4 Hz, H-6’), 6.89 (IH, d, J = 8,4 Hz, H-S), 6.42 (lH, A 3 =2 Hz, H-81, 6.22 (ZH, d, J=2 Hz, H-6)), S-19 (LH, d, J~7.5 Hz, H-l”), 35-4-U region (H-2”, H-3”, H-4”, H-5”, H&a and H&“b, overlapping signals). FAB MS (negative ion mode): m/z 463 [M -_K3-, 300 [M-H-163]-. Acknowledgement-We thank Dr Vincenzo De Feo (Dipartimento di Chimica delle Sostanze Naturali, Universiti “Federico II”, Napoh) for the collection and identification of plant material.
REFERENCES 1. Soukup, J. (1970) Vocabulurio de 10s nombres vulgpres de la jbra peruuna. Editorial Salesiana Lima. 2. Roersch, C. and van der Hoogte, L. (1988) Plantas Medicinales del Strrandim de1 Peru. Ed. Centro de Medicina Andina, Cuzco. 3. Markham, K. R., Tern& l3,, Stanky, R., Geiger, H. and Mabty, T, J. (1978) Tetrahedron 34, I389 4. Gorin, P. A. J. and Mazurek, M. (1975) Can. J, Ch?enr.53,1212. 5. Casy, A. F. (1971) PMR Spectroscopy in Mdicinal and Biological Chemistry, p. 30. Academic Press, New York. 6. Aquino, R., D’Agostino, M,, De Simone, F., Schettinq 0. and Pizza, C. (1987) B&&em. Sysf. &,I. 15,667. 7. De Feq V., DAgostino, M,, De Simone, F. and Pizza, C. (1990) Fitoterupia 5X,474. 8. D’Agostino, M., 3iagi, C,, De Feo, V., Zollo, F. and Pizza, C. (1990) Fitoterapia 61, 477.
12, pp. 43874388,
0031 9422/92 $5.00+0.00
1992
Pergamon PressLtd
FLAVONOL GLYCOSIDES FROM TAGEZ’ES ELLIPZTCA MARIOD'AGOSTINO,FRANCESCO
~~~irnento
DE SIMONE,ZHONG
LIANG ZHOU* and COSIMO PIZZA
di Chiiica delle Sostanze Naturali, Universiti di Napoli “Federico II”, Via Domenico Montesano, 49,80131 Napoli, Italy; *Shanghai Institute of Pharmaceutical Industry, Shanghai, People’s Republic of China (Received in revised form 16 April 1992)
Key Word Index-Tag&s elliptica; Asteraceae; quercetin 3-(3”,6“-diacetylgalactoside) and 3-(2”,3“,4”-triacetylgalactoside); flavonol glycosides.
Abstract-Two
new flavonol glycosides, quercetin 3-(~,~~~tylg~a~toside) and quercetin 3-(2”,3”,4”triacetylgalactoside) have been isolated from a methanolic extract of Tagetes elliptica and identified on the basis of chemical and spectral data. The known compounds quercetin 3-galactoside, isorh~e~ 3~rhamnosyl(l-*6)glucoside, myricetin 3&ucoside, querceti isorhamnetin 3-galactoside, quercetin 3-(6”-galloylgalactoside), quercetin 3rhamnoside, quercetin 3-rhamnosyl(l+Ci)galactoside and rhamnetin were identified from the same extract.
INTRODUCTION Tugetes eEl@ica Sm. is a herbaceous plant widespread in the Peruvian Sierra where it is used in folk medicine [i, 21. In an investigation of a methanoiic whole plant extract of T. elliptica two new flavonol glycosides (1 and 2) were isolated together with nine known compounds (3-11). The present paper describes the identification of 1 and 2. BESULTsANDDLSCUfSION
The ‘H NMR spectrum of l-3 showed the characteristic aglycone pattern of quercetin derivatives, namely a 2H AX and a 3H ABX system (see Experimental). The 13CNMR spectrum of these products exhibited all the upfield and downfield shiRs of the corresponding signals of quercetin consistent with 3-O-glycosidation [3]. The identity of quercetin as aglycone was further confirmed by W spectral data (see Experimental). The sugar moiety of 3 [3] was identified as B-Dgalactopyranose by FAB mass spectrometry, anomeric proton J value (7.5 Hz) and 13CNMR data [4] (see Table 1). However, the FAB mass spectrum of 1 showed singlets (3H) at 61.85 and 2.18 due to acetyl groups and diagnostic downfield shifts concerning H-3” (64.83, dd, J =9 and 4.2 Hz) and H-6”a (64.20, dd, J= 12 and 8.3 Hz). The corresponding signals for 3 [5] and anomeric coupling constant (J = 7.5 Hz) indicated that here the sugar moiety was 3,6-dia~tyl-~-D-g~~tose. This was confirmed by downfield shifts observed for the peaks of C-3“ (+ 2.3 ppm) and C-6” (+ 2.2 ppm) and by upfield shifts noted for the peaks of C-2” (- 2.8 ppm), C-4” (- 2.3 ppm) and C-S” ( - 4.0 ppm) in comparison with those of homologous carbons of 3 (see Table 1). The FAB mass spectrum and I-I-1” J value (7.5 Hz), along with the singlets (3H) at 6 1.86,2.05 and 2.16 showed that sugar residue of 2 was a ~-~t~a~tyl~iactose. The ‘HNMR spectrum indicated C-2”, C-3” and C-4” as acetylation sites, which was deduced by downfield shifts of H-4”, H-3” and H-2” signals in comparison with the co~es~nd~g signals for
3 (see Experimental) [S]. The identity of peaks was estab~shed on the basis of the multipIi~ty and decoupling experiments: irradiation at 85.01 (H-3”, dd, J =9 and 4.2 Hz) and 4.90 (H-2”, submerged by HOD signal) caused the simplification of signals centred, respectively, at 65.32 (H-4”, d, J=4.2 Hz) and 5.29 (H-l”, d, J = 7.5 Hz). The 2,3,4-triacetylation of a-D-g&iCtOSe was also consistent with i3CNMR data (see Table 1). The upfield shifts of C-l” (- 0.7 ppm) and C-5” (- 2.5 ppm) signais in comparison with those of the homologous carbons of 3 were ascribable, respectively, to acetyl groups linked at C-2” and C-4”. The total upfield shift (- 2.9 ppmf observed for C-3” and the chemical shifts of C-2” (670.6) and C-4” (668.9) were imputable to the combination of the acetylation downfield shift and upfield shift (y effect). Finally, 1 and 2 were both deacyiated to give 3, further confirming that gaiactose was the sugar moiety in these compounds. On the basis of these data, 1-3 were identified, respectively, as quercetin 3-O-~-D-(~‘,~-dia~tylgaI~topyranoside), quercetin 3-~-~-D-(2”,~,~-t~a~ty~g~actopyranoside) and the known quercetin 3-U+-galactopyranoside. Finally, the identification of known compounds isorhamnetin 3-rhamnosyl-( l-+6) glucoside (4), myricetin 3-glucoside (!5),quercetin (6), isorhamnetin 3-galactoside (7), quercetin 3~~-~lloylgal~toside (8), quercetin 3rhamnoside (9), quercetin 3-rhamnosyl-( 1-+6)galactoside (10) and rhamnetin (11) was based on comparison with literature data [3, 6-Q
EXPERIMENTAL FAB MS was carried out at 2-6 kV with a glycerolthioglycerol matrix in negative ion mode. The NMR spectra were carried out in CD,OD solutions at 250 MHz (IHNMR) and 62.5 MHz (‘“C NMR) with solvent as int. standard. Plant material. Tagetes elliptica was collected in Peru (province of Huacabamba, department of Piura) in October 1988. A voucher specimen is deposited in the Herbarium of Cattedra di
4387
Short Reports Table 1. “C NMR spectra of compounds CD,UD
Aglycone
c
1
2
3
2
158.5 135.7 179.5 163.0 loo.1 166.1 94.9 158.5 104.9 123.4 117.7 145.8 150.0 116.1 123.5 105.6 70.4 77.4 67,7 73.2 64*1 172.6 20.3 20.9
158.3 135.4 179.2 162.8 100.0 165.9 94.9 159.6 105.5 123.0 317.6 145.9 149.7 115.8 123.5 104.7 70.4 72.2 68.9 74.7 62.7 172.1 20.4 20.5 20.7
158.3 135.8 179.4 f 63.0 99,8 166.0 94.7 158.8 104,2 123.2 117.8 145.3 149.9 116.1 122.9 105.4 73.2 751 70.0 77.2 61.9
4 5 6
8 9 10
Galactow
C=O (Acetyl) Me (Acetyl)
l-3 in
(6)
2 3’ 4 5’ 6 1” 2” 3” 4” 5,I 6”
Botanica Farmaceutica (Dipartimento di Chimica delle Sostanze Naturali, Universitii “Fed&co IT”, Napoli). Extraction and isohtiun. The air-dried plant (400 g), defatted with petrol and CHCI,, was extracted with M&H at room temp. The crude extract (54 g), dispersed in H,U and reextracted with &OH, gave 16 g of a flavonoid mixture which was chromatographed on a Sephadex LH-20 column (80 x 4 cm) in 2-g batches. Fractions of 10 ml were eluted with MeQH, analysed by TLC (silica gel) in BuOH--HOAc-Hz0 (12: 3 : 5)and further fractionated by HPLC on a CIs p-Bondapak column (30 cm x 8 mm i.d., flow rate: 2.5 ml min- ’ ). Using MeOH-H&I I : 1as eluent, frs 9-15 (I.8 g) gave 4 f32 mg, R, 9 minb 10(66 mg R, 12 min), 1 (18 mg, R, 27 min) and 2 (24 mg, R, 41 min), whiL frs 33-40 (0.2 g) gave 6 (10 mg, R, 28 minf. With MeOH-H,O 2:3 as eluent, frs 16-22 (0.8 g) gave 3 (35 mg R, 30 min) and 9 (15 mg, R, 50 mm); frs 23-27 (0.2 g) gave 5 (14 mg, R, 19 min), 7 (10 mg, R, 29 min) and 11 (12 mg, R, 51 mm); frs 28-32 (0.3 g) gave 8 (34 mg, R, 19 mm). Alkaline /ry&oIysis. Compounds 1 and 2 (each 10 mg) in 0.5 M aq. RUH (1 ml) were heated at 110” in ;i stoppered reaction vial for 2 hr. The reaction mixture, adjusted to pH 7, was extracted with B&H. The organic phase was evapd to dryness, dissoived in CD,OD, and analysed by ‘H and t 3C NMR.
Quercetin 3-0-~-6-x)-(3”,5”-$i~~~~~~~~~~~~~~~~e~ (1). UV A,, nm: MeOH 256,302& 359; + AICI, 273,30&h, 436; + NaOAc 271, 317,413; NaOAc+H,BO, 258,293sh, 382; NaUMe 272, 321, 415. ‘HNMR: 67.80 (IH, i$ J=2 Hz, H-2’), 7.62 flH, dd, J=2 and 8.4 Hz, H-6’), 6.88 (lH, d, J =8.4 Hz, H-S), 6.41 (lH, d, J =2 Hz, H-B), 6.21 (lH, d, J=2 Hz, H-6), 5.22 (lH, d, Ja7.5 Hz, H-l”), 4.83 (lH, dd, J =9 and 4.2 Hz, H-3”), 4.20 (lH, dd, J = 12 and 8.3 Hz, 6”a), overlapping sign& in the 6 3.95-4.10 region (Hwb, H-2” and H-4”), 3.83 ffH, M, H-5”), 2.18 (3H, s, AC), 1.85 (3H, s, Ac). FAB MS (negative ion mode): m/z 547 fM -HI -, 504 [M-H-433-, 300 [M-H-247j-. Quercetin 3-o-B-o-(2”,3”,4”-triacetylgalactoside) (2). UV &laXnm: MeOH 257, 302sh, 358; +AlCl, 274, 30&h, 438; + NaOAc 272, 317, 414; +NaOAc+ H,BO, 258, 293sh 383; +NaOMe2?2,323,415. IHNMR: S?.65fIH,d,J=2 H&H-T), 7.50 (iH, dd, J =2 and 8.4 Hz, H-St), 6.87 (IH, d, J=8.4 Hz, HS), 6.35 flH, d, 5=2 Hz, H-81, 6.16 (IH, d, J=2 Hz, H-6), 5.32 (TN, Jz4.2 Hz, H-4”): 5.29 {lH, d, J=7.5 Hz, H-i”), 5.01 flH, dd, J=9 and 4.2 Hz, H-3”), 4.90 (H-2”, obscured by HOD), 3.90-4.10 region (H-5”, H-6”% H-6”b, overlapping signals), 2.16 (3H, s, Ac), 2.05 (3H, s, AC), 1.86 (3H, s, AC).FAR MS (negative ion mode): m/z 589 [M-H]-, 546 EM-H-43]-, 300 [M-H -239]-. Querceiirt ?-@&D-gnfuctoside (3). UV &,_ nm: M&H, 256* 301sh, 357; -I+AlCl, 274, 3OOsh,432; +NaOAc 273, 317, 393; NaOAc+H,BO, 254, 290, 377; NaOMe 273, 325, 412. ‘HNMR: 67.87 (lH, d, J=2 Hz, H-2’), 7.61 (lH, dd, J=2 and 8.4 Hz, H-6’), 6.89 (IH, d, J = 8,4 Hz, H-S), 6.42 (lH, A 3 =2 Hz, H-81, 6.22 (ZH, d, J=2 Hz, H-6)), S-19 (LH, d, J~7.5 Hz, H-l”), 35-4-U region (H-2”, H-3”, H-4”, H-5”, H&a and H&“b, overlapping signals). FAB MS (negative ion mode): m/z 463 [M -_K3-, 300 [M-H-163]-. Acknowledgement-We thank Dr Vincenzo De Feo (Dipartimento di Chimica delle Sostanze Naturali, Universiti “Federico II”, Napoh) for the collection and identification of plant material.
REFERENCES 1. Soukup, J. (1970) Vocabulurio de 10s nombres vulgpres de la jbra peruuna. Editorial Salesiana Lima. 2. Roersch, C. and van der Hoogte, L. (1988) Plantas Medicinales del Strrandim de1 Peru. Ed. Centro de Medicina Andina, Cuzco. 3. Markham, K. R., Tern& l3,, Stanky, R., Geiger, H. and Mabty, T, J. (1978) Tetrahedron 34, I389 4. Gorin, P. A. J. and Mazurek, M. (1975) Can. J, Ch?enr.53,1212. 5. Casy, A. F. (1971) PMR Spectroscopy in Mdicinal and Biological Chemistry, p. 30. Academic Press, New York. 6. Aquino, R., D’Agostino, M,, De Simone, F., Schettinq 0. and Pizza, C. (1987) B&&em. Sysf. &,I. 15,667. 7. De Feq V., DAgostino, M,, De Simone, F. and Pizza, C. (1990) Fitoterupia 5X,474. 8. D’Agostino, M., 3iagi, C,, De Feo, V., Zollo, F. and Pizza, C. (1990) Fitoterapia 61, 477.