[Agr. Biol. Chem., Vol. 34, No. 6, p. 900•`907, 1970]

Isolation and Structure of New Flavonoids, Flavoyadorinin-A.

Flavoyadorinin-B and Homo-flavoyadorinin-B, in the Leaves

of Viscum album Linnaeus var. coloratura Ohwi

Epiphyting to Pyrus communis Linnaeus_??_

By Naokazu OHTA and Kazuyoshi YAGISHITA

Department of Food Chemistry, Kumamoto Women'sUniversity, Ooemachi, Kumamoto City, Japan

Received November 10, 1969

The flavonoid constituents of the leaves of Viscum album Linnaeus var. coloratum Ohwi

(Viscaceae) epiphyting to Pyrus communis Linnaeus were examined. Three kinds of new flavonoids named Flavoyadorinin-A (I), C23H24O12, mp245•`246•Ž, Flavoyadorinin-B (II),

C23H24O11•E3H2O, mp 203•`206•Ž, and Homo-flavoyadorinin-B (III), C28H32O15•E3H2O, mp 208•`210•Ž (sint. at 130•Ž) were isolated from the leaves of this plant. The structures of the Compound (I), (II), and (III) were showed to be 7,3'-di-O-methyl-

-3-O-mono-D-glucoside (-3-0-mono-D-glucoside), 7,3'di-O-methyl-luteolin- 4'-O-mono-D-glucoside, and 7,3'-di-O-methyl-luteolin-4'-O-D-glucoapioside, respectively.

Japanese mistletoe, Viscum album Linnaeus mistletoe (Viscum album Linnaeus var. coloratum var. coloratum Ohwi is a perennial shrub of Ohwi) epiphyting to Pyrus communis Linnaeus. Viscaceae, which grows wild in Saghalien, As described in the experimental part, Kurles, Honshu, Shikoku, Kyushu, Formosa, three flavonoids were isolated in pure state Korea and China. on a cellulose-column chromatography, Com-

In 1957, the isolation of four kinds of pound (I), mp 245•`246•Ž, Compound (II), flavonoids, Compound A, mp208•`210•Ž mp203•`206•Ž, and Compound (III), mp (positive Molish reaction), Compound B, mp 208210°C. All of these compounds, (I), 107•`109•Ž, Compound C, mp 142•`143•Ž, (II) and (III), were new flavonoids, which and Compound D, mp219•`220•Ž (all were would be named Flavoyadorinin-A Flavo positive to Mg-HCl reaction) were reported yadorinin-B, and Homo-flavoyadorinin-B, re by Ts'eng Kuang-Fang et al .1) No further spectively. investigations of their structures, however, The compound (I), named flavoyadorinin-A, have been made. was obtained as pale yellowish needles, mp The present paper deals with the isolation 245246°C, and from its analytical values and the structural elucidation of flavonoid was suggested the formula C23H24O12 contain compounds in the fresh leaves of Japanese ing two methoxyl groups. It exhibited a dark green color with ferric chloride, and _??_A part of this work was presented at the General the reduction tests for flavonoid (Mg-HCl, Meeting of the West Japan Branch of the Agricultural Zn-HCl) were positive. It also was positive Chemical Society of Japan, held on October 20, 1968, at the Shimane University, Japan. in the Gibbs indophenol test. The UV- 1) Ts'eng Kuang-Fang and Li Shil-Chuen , Yao spectrum of this compound (I), maxima Hsueh Hsueh Pao (Acad. Sinica), 5, 169 (1957). 253.4, 268, and 350 mfg, and its pattern, Isolation and Structure of New Flavonoids, Flavoyadorinin-A, Flavoyadorinin-B 901

were very similar to those reported for a (Mg-HCl, Zn-HCl) were positive. It was also number of flavonoids of the quercetin type. positive in the Gibbs test. The UV-spectrum This was also supported by the fact that of this compound (II), maxima 2436, 2712, demethylation of the compound (I) with and 334•`6mƒÊ, and its pattern, were very hydriodic acid in phenol gave quercetin, mp similar to those reported for a number of 314•`317•Ž, which was further characterized flavonoids of the luteolin type. This also as its penta-acetate (mp201•Ž). was supported by the fact that demethylation On the hydrolysis with 30o sulfuric acid, of the compound (II) with hydriodic acid in the compound (I) gave an aglycon (IV) as pale phenol gave luteolin, mp 326•`329•Ž. Its yellowish needles (mp 2l0•`216•Ž, 66.79%), acetate, mp 226•`227•Ž. which was identical with the authentic On the hydrolysis with 3% sulfuric acid specimen of rhamnazin (7,3'-di-O-methyl- the compound (II) gave a new aglycon (V) as quercetin)* obtained from the leaves of Poly yellowish granular crystals (58.51%), C15H8O4•E gonum hydropiper Linnaeus. From the hy (OCH3)2, mp 218•`220•Ž, which would be drolyzate freed of the aglycon (IV), D-glucose named flavoyadorigenin-B, hereafter. Acetate was proved to be present by paper chro of this aglycon (V), mp 182•`186•Ž. In the matography. Alkali fusion of the compound hydrolyzate freed of the aglycon (V), only (I) under the usual manner gave mono-O- D-glucose was detected as the sugar component methyl-phloroglucinol and vanillic acid. The by paper chromatography. Alkali fusion of compound (I) and its aglycon (IV) were the compound (II) under the usual manner confirmed by comparing their UV-spectra gave mono-O-methyl phloroglucinol, and with those in presence of the following vanillic acid. In the UV-absorption deter reagents:" 1) sodium acetate (both the com- mination, the following results were con

pound (I) and its aglycon (IV):-no change); firmed: 1) both the compound (II) and its 2) aluminum chloride (the compound (I):-no aglycon (V) showed no shift of the absorption change, its aglycon (IV):-the bathochromic maximum by adding sodium acetate, suggest- shift of 59 mg); and 3) sodium ethylate (the ing the presence of 7-methoxyl group, and compound (I):-the long wavelength bands also showed a significant bathochromic shift

disappearing, its aglycon (IV):-the batho (about 10mƒÊ) by adding aluminum chloride, chromic shift of 57 mp). indicating the presence of 5-hydroxyl group On the basis of these data the structure of in their nucleus; 2) the aglycon (V) showed the compound (I) might be rhamnazin-3-O- a characteristic bathochromic shift (70mƒÊ) of mono-D-glucoside. absorption maximum by adding sodium

The compound (II), named flavoyadorinin- ethylate, suggesting the presence of 4'- B, was obtained as yellowish needles, mp hydroxyl group in its structure.

203-206°C, and from its analytical values From these results the structure of the

was suggested the formula C21H18O9(OCH3)2•E compound (II) and its aglycon (V) might be

3H2O. It gave brown color with ferric chlo 7,3'-di-O-methyl-luteolin-4'-O-mono-D-glucoside

ride, and the reduction tests for flavonoid and 7,3'-di-O-methyl-luteolin, respectively.

The compound (III), named homo-flavo

* The authentic sample of rhamnazin was fur yadorinin-B, was obtained as yellowish needles, nished through the courtesys of Dr. M. Aritomi mp 208•`210•Ž (sint. at 130•Ž), and analyzed (Kumamoto University) and Dr. A. Kawase (Hiroshima for C26H26O13(OCH3)2•E3H2O. It gave brown Women's University). color with ferric chloride, and the reduction 2) L. Jurd, "The Chemistry of Flavonoid Com- tests for flavonoids were positive. Gibbs: pounds," ed. by T. A. Geissman, Pergamon Press, 1962, p. 107. positive. The UV-spectrum of the compound 902 N. OHTA and K. YAGISHITA

() E. Wada, J. Am. Chem. Soc., 78, 4725 (1956). () A. G. Perkin and J. R. Allison, J. Chem. S oc., 81, 468 (1902). () R. Chatterjee and D. K. Datt, Indian J. Physiol., 4, 61 (1950). () A. G. Perkin and P. J. Wood, J. Chem. Soc., 73, 374 (1898). (rhamnazin) A. G. Perkin and J. Geldard, ibid., 67, 496 (1895). () G. B. Marini-Bettolo, V. Deulofeu and E. Hug, Gazz. chim. ital., 80, 63 (1950). () F. E. King, T.J. King and K. Sellars, J. C mem. Soc., 1952, 92.

(diosmetin) E. Vongerichten, Ber., 33, 2334 (1900). (chrysoeriol) F. B. Power and F. Tutin, Proc. Am. Pharm. Assoc., 54, 352 (1906).

(III), maxima 243---6, 271- 2, and 340 mp, and Rf value with that of D-glucose and the other its pattern, were almost the same as that of was also proved to be apiose, by cochromato the compound (II), suggesting a close simi graphy on paper using the sugar solution larity in the structure between the compound obtained from the hydrolyzate of authentic (II) and the compound (III). This was also apiin.* Moreover, partial hydrolysis of the proved by the fact that the compound (III) compound (III) with 5% acetic acid gave the with hydriodic acid in phenol in the same compound (II) and apiose. manner as before gave luteolin, mp 326 The compound (III) was also confirmed by •` 329•Ž. comparing its UV-spectrum with those in On the hydrolysis with sulfuric acid in the presence of the following reagents: 1) sodium usual manner, the compound (III) produced acetate (no change), 2) aluminum chloride an aglycon as yellowish granular crystals (the bathochromic shift of about 10 mp), 3) (50.1%), C15H8O4(OCH3)2 mp 218•`220•Ž, sodium ethylate (the long wavelength bands which was proved to be identical with the disappearring). compound (V), by mixed mp and IR-absorp Consequently, the structure of the com- tion data. In the hydrolyzate freed of the

aglycon (V), two kinds of sugar components * The authentic specimen of apiin was furnished were proved to be present by paper chro through the courtesy of Dr. A. Kawase (Hiroshima matography. One of these was identical in Women's University). Isolation and Structure of New Flavonoids, Flavoyadorinin-A, Flavoyadorinin-B 903

pound (III) might be 7,3'-di-O-methyl-luteolin- that flavonoid compounds in the leaves of 4'-O-D-glucoapioside. Viscum album Linnaeus var. coloratura Ohwi In this way, it has now become evident epiphyting to Pyrus communis Linnaeus consist of three kinds of 7,3'-di-O-methyl flavonoids, i.e., flavoyadorinin-A (I), flavoyadorinin-B (II), and homo-flavoyadorinin-B (III). The ratio of the above mentioned flavonoid contents, i.e., the compound (I) to (II) and to (III) was about 4: 1 : 60. In addition, it is chemotaxonomically inter esting to note that from the same plant source epiphyting to the different host plant, i.e. Zelkowa serrata Makino, only two kinds of flavonoids, i.e., flavoyadorinin-B (1I) and homo-flavoyadorinin-B (III) have been proved to be present by paper chromatographical examination.

EXPERIMENTAL

All melting points were uncorrected. Ultraviolet spectra were measured after Jurd,2) by using a Hitachi EPS-2U recording spectrophotometer, and infrared spectra in KBr discs by using a Nihon Koken DS-301 spectrophotometer.

Paper chromatography. The flavonoid compounds FIG. 1. Infrared Absorption Spectra of a: Flavoya were run in the following solvent systems by the dorinin-A (I), b: Its Aglycon (IV), c: Flavoya ascending method: (I) BuOH-AcOH-H2O (4:1:5, dorinin-B (II), d: Homo-flavoyadorinin-B (III), e: upper phase) (solv. 1), (II) 60% AcOH (solv. 2), (III) Flavoyadorigenin-B (V), and f: Its Acetate (in 30% AcOH (solv. 3), and (IV) 15% AcOH (solv. 4). KBr). A spray reagent used 3% sodium carbonate solution

TABLE I. ULTRAVIOLET ABSORPTION SPECTRA (ƒÉmax mƒÊ (log ƒÃ))

a) shoulder 904 N. OHTA and K. YAGISHITA

or 1% ferric chloride ethanolic solution. The com containing yellowish needles (Substance C) (1.2g, 0.0280), mp 197•`202•Ž (sint. at 130•Ž). ponent sugar was run in the following solvent systems: solv. 1, (V) BuOH-pyridine-H2O (3:2:1.5) (solv. 5), Another half volume of the resulting aqueous solu

(VI) PhOH-H20 (5:1) (solv. 6), (VII) BuOH-EtOH- tion was treated by the same procedure descrived H20 (4:1:5, upper phase) (soly. 7), by ascending above. Yield of the total Substance B and C, were

method, and detected by spraying with the following 130 mg and 2.3 g, respectively. reagents: benzidine-trichloroacetic acid, diphenyl amine-aniline, aniline-phthalic acid, naphthoresorcinol- Isolation of the compound (I) (flavoyadorinin-A). The

phosphoric acid or silver nitrate reagents. The Substance A (400 mg), mp 232•`240•Ž, was recrys phenolic and acidic compounds were run in the tallized several times from ethanol, giving pale solvent of BuOH saturated with 20 ammonium yellowish needles (370 mg) mp 245•`246•Ž. It gave hydroxide (sole. 8) and detected by spraying with the following color reactions: ferric chloride: dark

diazotized sulfanilic acid. brown; sodium carbonate: yellow; basic lead acetate:

yellow; Mg-HCl: yellow; Zn-HCl: orange red. Extraction and isolation of pigments. The fresh Under UV-light: brown. Gibbs: positive. This leaves (4.2 kg)* were extracted with boiling ethanol compound is soluble in 30•`80% ethanol or methanol, and the extract was condensed to a syrup in vacuo. ethyl acetate, and pyridine, hardly soluble in ethanol The syrupy substance was digested with boiling water and methanol, and insoluble in chloroform, benzene, and the resulting aqueous solution was repeatedly ethyl ether, petroleum ether, acetone, hexane and shaken with chloroform, giving a reddish brown water. The Rf values: 0.28 (solv. 1); 0.81 (solv. 2); aqueous solution, whereby a pale yellowish gray color 0.80 (solv. 3); 0.72 (solv. 4). ƒÉ_??_mƒÊ (log ƒÃ): 253-4 substance (4.1 g, ferric chloride and sodium carbonate: (3.44), 268 (3.40), 350 (3.36). Anal. Found: C, 50.23; both negative) was observed between chloroform and H, 5.44; OCH3, 11.22. Calcd. for C21H18O10(OCH3)2•E aqueous layers. The yield of the chloroform soluble 3H2O: C, 50.54; H, 5.49; OCH3, 11.35%. fraction** was 121.O g (2.850 of the fresh leaves). The aqueous solution freed of the chloroform soluble Hydrolysis of the compound (1). The compound (I) fraction was further extracted with ethyl acetate (40.5 mg) was refluxed with 5% sulfuric acid in 50% exhaustively, giving u reddish brown extract (4 spots ethanol solution for 3 hr. After removing the ethanol in sole. 1; Rf values: 0.88, 0.73, 0.48, and 0.28). in vacuo, the resulting solution was diluted with a After removing the solvent, the ethyl acetate extract small amount of water, and was extracted with was dissolved in a suitable amount of ethanol and ethyl acetate several times. The ethyl acetate soluble was left to stand in a refrigerator for a week or fraction were combined ,together, washed with a more. The pale yellowish granular crystals (Substance small amount of water, and then evaporated off. A), mp 232•`240•Ž, gradually separated were collected The residue was recrystallized several times from by filtration. Yield, 400 mg (0.0095%). ethanol giving pale yellowish fine needles (IV) (27.1 The remaining filtrate freed of the Substance A mg, 66.79%), mp 210-216°C either alone or on was dried up to remove the solvent, and was dissolved admixture with an authentic specimen of rhamnazin in a suitable amount of water. One half of this isolated from the leaves of Polygonum hydropiper resulting aqueous solution was chromatographed on a Linnaeus.* Rf values: 0.85 (solv. 1), 0.53 (solv. 2), column of cellulose (3.0 x30cm). Elution with water- 0.10 (solv. 3), and 0.05 (tailing) (solv. 4), were found saturated ethyl acetate of the column gave a fraction to be the same as those of authentic rhamnazin. (yellow color) containing yellowish needles (Substance ƒÉ_??_mƒÊ (log ƒÃ): 255.6 (4.44), 3723 (4.42). Anal. B) (67 mg, 0.0015%), mp 194•`200•Ž. The column Found: C, 61.52; H, 4.43; OCH3, 18.35. Calcd. for freed of the Substance B was further eluted with C15H8O5(OCH3)2: C, 61.81; H, 4.23; OCH3, 18.78%. ethanol, giving a fraction (yellowish brown color) Hydrolyzate freed of the compound (IV) was neutralized with barium carbonate, and filtered to

* The fresh leaves were collected at Nango -dani, yield a clear solution, which was used for detection Mt. Aso in the spring of 1967. of the sugar component. D-Glucose was proved to ** In this chloroform soluble fraction be the sugar component of the compound (I) by , oleanolic acid, ƒÀ-amyrin, lupeol and myristic acid have been paper chromatography (solv. 5) using authentic D- proved to be present. glucose as check. Isolation and Structure of New Flavonoids , Flavoyadorinin-A, Flavoyadorinin-B 905

Demethylation of the compound (I); Formation of quercetin . solution was left to stand in a refrigerator over night. A mixture of the compound (I) (110mg), hydriodic The aglycon separated was collected, washed with acid (4ml), and phenol (6 ml) was heated at 100•Ž ice-cooled water, and recrystallized from ethanol in nitrogen stream for 1.5 hr. After cooling, the giving yellowish fine needles (V) (23.6 mg, 58.510), reaction mixture was diluted with a suitable amount mp 220°C. Color reactions of this compound (V): of water, and treated with sodium bisulfate. A ferric chloride: brown; sodium carbonate: yellow; crystalline substance was obtained, which was crys basic lead acetate: yellow; Mg-HCl: orange red: Zn- tallized from ethanol as yellowish needles (45 mg), HCl: orange yellow. Its solubilities: soluble in 30•` mp 314•`317•Ž either alone or on admixture with 100% ethanol or methanol, acetone, and pyridine; an authentic specimen of quercetin. Acetylation of insoluble in chloroform, benzene, ethyl ether, petro this demethylated compound with acetic anhydride- leum ether, hexane and water. Rf values: 0.92 (solv. pyridine in the usual manner gave almost colorless 1), 0.76 (solv. 2), 0.3 (tailing) (solv. 3), 0.09 (tailing) needles, mp 200•`201•Ž, which were identical with (solv. 4). ƒÉ_??_mƒÊ (logƒÃ): 251-3 (4.20), 268 (4.16), authentic quercetin pentaacetate (mp201•Ž). 350 (4.31). Anal. Found: C, 70.03; H, 4.75; OCH3,

21.05. Calcd. for C15H8O4(OCH3)2: C, 70.34; H, Alkali fusion of the compound (I). A mixture of 4.82; OCH3, 21.37%.

the compound (I) (24 mg) and 20,06 potassium hy Hydrolyzate freed of the aglycon (V) was neutral droxide solution (10 ml) was heated at 120°C in ized with barium carbonate, and filtered to yield a nitrogen for 30 min. After cooling and diluting with solution, which was used for detection of the sugar

water, the resulting reaction mixture was acidified component. D-Glucose was proved to be the sugar with dil. hydrochloric acid and extracted with ether. component of the compound (II) by cochromatography The ethereal extract was subjected to paper chro on paper.

matography (solv. 8). Two spots were revealed on the chromatogram, the Rf values of which agreed Acetylation of the compound (V). To a solution of

with those of authentic mono-O-methyl phloroglucinol the compound (V) (25 mg), mp 220°C, in pyridine

(Rf: 0.88, blood color) and vanillic acid (Rf: 0.09, (1 ml) was added acetic anhydride (1.5 ml), and the reddish brown color), respectively. mixture was allowed to stand at room temperature for 2---3 days. The reaction mixture was poured Isolation of the compound (II) (flavoyadorinin-B)). The into ice-cooled water, and the crude acetate separated

Substance B (130 mg), mp l94•`200•Ž, was recrys was filtered, washed with water, and then dried. tallized several times from 50% ethanol, giving pale After recrystallizations twice from ethanol, the pure

yellowish needles (112 mg), mp 203•`206•Ž. It gave acetate obtained as almost colorless needles (20 mg), the following colorations: ferric chloride: purplish mp 182.-186°C. Color reactions: ferric chloride:

brown; sodium carbonate: yellow; basic lead acetate: negative; sodium carbonate: yellow; basic lead acetate:

yellow; Mg-HCl; orange; Zn-HCl: orange red. pale yellow. Under UV-light: fluorescence. Its solu Under UV-light: brown. Gibbs: positive. This com- bilities: easily soluble in chloroform, hexane, dioxane

pound is soluble in 30-80% ethanol or methanol, and pyridine; soluble in ethanol, methanol, acetone, water-saturated ethyl acetate, and pyridine, hardly benzene, ethyl acetate, ethyl ether and petroleum soluble in ethanol, methanol, and ethyl acetate, and ether; and insoluble in water. Anal. Found: C, 63.07;

insoluble in chloroform, benzene, ethyl ether, petro H, 4.67; OCH3, 15.15. Calcd. for C15H6O4(OCH3)2

leum ether, acetone, hexane and water. Rf values: (CH3CO)2: C, 63.31; H, 4.52; OCH3, 15.57%. 0.76 (solv. 1); 0.83 (solv. 2); 0.62 (solv. 3); 0.33

(solv. 4). ƒÉ_??_mƒÊ (logƒÃ): 243•`6 (4.21), 2712 Demethylation of the compound (II); Formation of luteolin. A mixture of the compound (II) (51 mg), (4.28), 334•`6 (4.31). Anal. Found: C, 55.07; H, 5.66; OCH3, 11.62. Calcd. for C22H18O9(OCH3)2•E hydriodic acid (4 ml), and phenol (6 ml) was heated

3H2O: C, 55.14; H, 5.51; OCH3, 11.69%. at 100°C in nitrogen for 1.5 hr. After cooling the reaction mixture was worked up as described before. Hydrolysis of the compound (H); Formation of flavo The demethylated compound obtained was crystallized yadorzgenin-B (V). The compound (II) (40.3 mg) was from ethanol as yellowish needles (22 mg), mp 325•` refluxed with 50 sulfuric acid in 50% ethanol for 327•Ž either alone or on admixture with an authentic 3 hr. After removing ethanol in vacuo, the resulting specimen of luteolin. Acetylation of this demethylated 906 N. OHTA and K. YAGISHITA

TABLE II-1. Rf VALUES OF SUGARSAFTER HYDROLYSIS

TABLE 11-2. Color REACTIONS OF SUGARS ethanol, giving pale yellowish needles (2.15g), mp PRESENT IN HYDROLYZATE 208•`210•Ž (sint. at 130•Ž). It gave the following colorations: ferric chloride: purplish brown; sodium

carbonate: yellow; basic lead acetate: yellow; Mg- HCl: orange; Zn-HCl: orange red. UV-light: brown.

Gibbs: positive. Its solubilities: soluble in 30-80%

ethanol or methanol, ethyl acetate, and pyridine; hardly soluble in ethanol and methanol; and insoluble in acetone, chloroform, benzene, ethyl ether, petro

leum ether, hexane and water. Rf values: 0.46 (solv. 1); 0.90 (solv. 2); 0.75 (solv. 3); 0.49 (solv. 4). ƒÉ_??_ mƒÊ (logƒÃ): 2434 (4.39), 270-1 (4.32), 340 (4.36).

Anal. Found: C, 51.47; H, 5.72; OCH3, 9.07. Calcd.

for C26H26O13(OCH3)2•E3H2O: C, 51.77; H, 5.29; OCH3, 8.93%. a) R. W. Bailey and E. J. Boure, J. Chromato., 4, 206 (1960).b) Hydrolysis of the compound (III); Formation of the S. M. Partidge, Nature, 164, 443 (1949).c compound (V) and detection of apiose. The compound

) V. Prey, H . Berbalk and M. Kausz, Mikrochim. (III) (50 mg) was refluxed with 5% sulfuric acid in Acta, 1961, 968. 50% ethanol for 3 hr. After removing ethanol in d) W . E. Trevelyan, D. P. Procter and J. B. Har vacuo, the resulting solution was treated as described rison, Nature, 166, 444 (1950). before. The aglycon separated was recrystallized several times from 80% ethanol, giving yellowish compound with acetic anhydride-pyridine in the usual fine needles (25.0 mg, 50.1%), mp 218-220°C, which manner gave almost colorless needles, mp 224227°C, was proved to be identical with the compound (V) which were identical with an authentic specimen of by mixed mp and IR spectrum. luteolin-tetraacetate, mp 226•`227•Ž . Hydrolyzate freed of the aglycon was neutralized

Alkali fusion of the compound (II). A mixture of with barium carbonate, and filtered to yield a solution, the compound (II) (19mg) and 20% potassium hy which was subjected to paper chromatography for droxide solution was heated at 120•Ž in nitrogen for detection of the sugar component. Two spots were 30 min. After cooling and diluting with water detected on the chromatogram, the Rf values of , the resulting mixture was acidified with dil. hydrochloric which respectively agreed with those of D-glucose and acid and extracted with ether . The ethereal extract apiose obtained from the hydrolyzate of authentic apiin* (Table II). was subjected to paper chromatography (solv . 8). Two spots were revealed on the chromatogram , the Rf values of which agreed with those of authentic mono- Partial hydrolysis of the compound (III); Formation of O-methyl phloroglucinol and vanillic acid the compound (11). A suspension of the compound , respectively. (III) (a few mg) in 5% acetic acid (10 ml) was heated Isolation of the compound (III) (homo-flavoyadorinin-B). on a water-bath for 3 hr. After cooling and diluting

The Substance C (2.3g), mp l97•`202•Ž (sint . at with a small amount of water (ca. 5 ml), the result- 130•Ž), was recrystallized several times from 50% ing reaction solution was subjected to paper chro- Isolation and Structure of New Flavonoids, Flavoyadorinin-A, Flavoyadorinin-B 907 matography using solv. 1•`4 (for flavonoid component) thanks to Dr. S. Kuwatsuka of Rikagaku-Kenkyusho and solv. 1 and 5•`7 (for sugar component). Two (Tokyo), for carrying out elementary analyses, to Mr. components were detected on the chromatograms, Y. Shimada of Kenmotsudai Arboretum of Kumamoto the Rf values of which agreed with those of the Forestry Bureau (Kumamoto), for determining the compound (II) and apiose, respectively (Table II). proper classification of the plant, and to Mr. M. Kitamura for his help in collecting the plant materials

Acknowledgement. We would like to express our used in this work.