Journal of the Korean Wood Science and Technology

The Korean Society of Wood Science & Technology

2020; 48(6):888-895

pISSN: 1017-0715, eISSN: 2233-7180

DOI: https://doi.org/10.5658/WOOD.2020.48.6.888

Original Article

Chemotaxonomic Significance of Catechin 7-O-beta-D-apiofuranoside in Ulmus Species

Mi KIM²^,*, Yong Jo LEE³^,*, Jae-Cheon SHIN⁴, Sun Eun CHOI⁵^,^†

²Department of Cosmetology Science, Nambu University, Gwangju 62271, Republic of Korea

³YEONGHWA FEED CO., LTD., #311, Chosun University Incubator Center, 309, Pilmun-daero, Dong-gu, Gwangju 61452, Republic of Korea

⁴Pohang Center for Evaluation of Biomaterials, 394, Jigok-ro, Nam-gu, Pohang, Gyeongbuk 37668, Republic of Korea

⁵Division of Forest Material Science & Engineering, College of Forest & Environmental Sciences, Kangwon National University, Chuncheon 24341, Republic of Korea

These authors contributed equally to this work.

^†Corresponding author: Sun Eun CHOI (e-mail: oregonin@kangwon.ac.kr, ORCID: 0000-0002-6306-0372)

Received: Sep 11, 2020; Accepted: Nov 02, 2020

Published Online: Nov 25, 2020

Abstract

Ulmus genus has excellent various physiological activities, including anti-ulcer, antioxidant, antibacterial, anti-cancer, immunity, and homeostasis maintenance effects, and it is known to have many additional drug effects And one of reasons for these excellentbiological activities is a flavan-3-ol chemical group in Ulmus genus. In this study a new flavan-3-ol compound was identified in Ulmus davidiana var. japonica. A flavan-3-ol,(2R,3S)-7-[(2S,3R,4R)-3,4-dihydroxy-4-(hydroxymethyl)oxolan-2-yl]oxy-2-(3,4-dihydroxyphenyl)-3,4-dihydro-2H-chromene-3,5-diol, named as catechin 7-O-beta-D apiofuranoside, was isolated from the stems and barks of Ulmus davidiana var. japonica for. suberosa, which is a species belonging to the genus Ulmus, growing throughout the Korea peninsula. The structure was elucidated by various spectroscopic methods including high-resolution TOF mass spectrometry, ¹H-NMR and ¹³C-NMR spectrometry and comparison with chemical structures of defined compounds.

Keywords: Ulmus davidiana var. japonica for. suberosa; flavan-3-ol; chemotaxonomy

1. INTRODUCTION

More than 30 species belonging to Ulmus genus grow in temperate regions of the Northern Hemisphere. Six species and two varieties of this genus are found in Korea: U. davidiana Planchon, U. davidiana Planchon var. japonica Nakai, U. laciniata Mayr, U. macrocarpa Hence, U. macrophylla Nakai, U. parvifolia Jacq.(U. coreana Nakai), U. pumila Linne (U. mandshurica Nakai) and Ulmus davidiana var. japonica for. Suberosa (Jung, 1974; Lee, 1996).

Flavan-3-ols are characteristic chemical components of Ulmus species (Son et al., 1989; Moon et al., 1995; Doskotch et al., 1973; Jung et al., 2007). Several interesting biological activities of flavan-3-ols including anti-inflammatory, anti-cancer, analgesic, antimicrobial, antineoplastic, anti-ulcer, antioxidant, antibacterial, anticancer and immunomodulatory properties have been reported previously (Hong et al., 1990; Park et al., 1996; Yang et al., 1996; Cho et al., 1996; Jun et al., 1998; Lee et al., 2000; Lee et al., 2001; Kim et al., 2002).

A flavan-3-ol compound, (+)-catechin 7-O-β.-D-xy-lopyranoside was isolated from the bark of U. Americana (Doskotch et al., 1973). In addition, (+)- catechin and (+)-catechin 5-O-β.-D-apiofuranoside from roots of U. davidiana were isolated (Son et al., 1989). Isoquercetin and rutin from leaves of U. parvifolia were isolated (Kim et al., 1992). Sterols, sterol glucoside and (+)-catechin 7-O-α-L-rhamnoside from barks of U. parvifolia were isolated (Moon et al., 1995). Sesquiterpene O-naphthoquinone of davidianone A, B, C and mansonone E, F, H, I from roots of U. davidiana were isolated (Kim et al., 1996). (+)-Catechin7-O-β.-Dapiofuranoside, lyoniside, 5’-methoxyisolariciresinol-9’- O-β.-D-xylopyranoside, rel-trans dihydrodehydroconiferyl alcohol 4-O-α-L-rhamnoside, icariside and ulmicine A-E from barks and roots of U. davidiana var. japonica were isolated (Bae et al., 2000), respectively.

In this study, we isolated and identified catechin 7-O-beta-D apiofuranoside from the stems and barks of U. davidiana var. japonica for. suberosa. We also report the screening of catechin 7-O-beta-D apiofuranoside from other Ulmus species [U. davidiana Planchon (leaves, stems and barks), U. davidiana var. japonica (leaves, stems), U. parvifolia Jacq (stems and barks) and U. pumila Linne (stems)].

2. MATERIALS and METHODS

2.1. Plant extracts of Ulmus species

The stems and barks of U. davidiana var. japonica for. suberosa (bar code; PB2437.1) as well as MeOH extracts of U. pumila Linne (stems, bar code; PB2430.2), U. parvifolia Jacq (stems and barks, bar code; PB2427.1), U. davidiana var. japonica (stems, leaves, bar code; PB2436.6~7), and U. davidiana Planchon (leaves, stems and barks, bar code; PB2435.1~2) were purchased from the Korea Plant Extract Bank(Cheongju, Korea).

2.2. Chemical separation and identification

Thin layer chromatography (TLC) was carried out using a pre-coated silica gel 60 F₂₅₄ plate (Merck, Darmstadt, Germany) with chloroform, methanol and water (70:30:4, volume ratio). The spots were detected under UV radiation (254 nm) and by spraying with FeCl₃ and 10% H₂SO₄ followed by heating.

The components derived from the Ulmus species were identified by several instrumental analyses. Based on 1D nuclear magnetic resonance (NMR), ¹H-(400MHz) and ¹³C-(100MHz) NMR experiments were recorded with FT-NMR Spectrometer 400 MHz, AVANCE III HD 400 (Bruker Corporation, Billerica, Massachusetts, USA) at Chonnam National University. Also, a high-resolution TOF mass spectrometry was conducted with JMS-T200GC (JEOL, Tokyo, Japan) at the National Center for Inter-University Research Facilities on Chonnam National University.

2.3. Quantitative analysis of catechin 7-O-β-D apiofuranoside of Ulmus species using high pressure liquid chromatography (HPLC)

HPLC was used for the quantitative analysis of catechin 7-O-beta-D apiofuranoside. A Waters 2695 separation module (Milford, MA, USA) equipped with a vacuum degasser, a binary pump, a 2487 dual λ absorbance detector and column compartment, was used to separate 1 on a Phenomenex KJ0-4282 guard column and SkyPakC18 column (4.6 × 250 mm, 5 ìm particle) with a linear gradient [D.W. : MeOH : P₂HO₄ (940 : 50 : 1)] : [MeOH : P₂HO₄ (990 : 1)] = 100 : 0 to 0 : 100 for 30 min. The column temperature was maintained at room temperature. The flow rate was 1.0 mL/min. The injection volume of each sample was 20 μL. The system was monitored at 280 nm (λ_max of catechin 7-O-beta-D apiofuranoside) eluting at14.44 ± 0.14 min, and catechin 7-O-β-D apiofuranoside was detected in the extracts of the stems and barks of U. davidiana var. japonica for. suberosa, as well as the extracts of U. davidiana Planchon (leaves, stems and barks), U. davidiana var. japonica (leaves, stems), U. parvifolia Jacq (stems and barks) and U. pumila Linne (stems).

3. RESULTS and DISCUSSION

Brown amorphous powder, high-resolution TOF MS m/z: 422.17511 [M]⁺; ¹H-NMR (400 MHz, DMSO-d₆ +D₂O): 6.74 (H-2’, 1H, d, J = 2.0 Hz), 6.69 (H-5’, 1H, d, J = 8.4 Hz), 6.59 (H-6’, 1H, dd J = 2.0, 8.4 Hz), 6.09 (H-8, 1H, d, J = 2.4 Hz), 5.90 (H-6, 1H, J = 2.4 Hz), 5.33 (H-1’’, 1H, d, J = 4.0 Hz), 4.55 (H-2, 1H, d, J = 7.2Hz), 4.03 (H-2’’, 1H, d, J = 4.0 Hz), 4.00 (H-4a’’, 1H, d, J = 9.6Hz), 3.89 (H-3, 1H, m), 3.68 (H-4b’’, 1H, d, J = 9.6 Hz), 3.45 (H-5’’, 2H, m), 2.65 (H-4a, 1H, dd, J = 4.8, 16.0Hz), 2.40 (H-4b, 1H, dd, J = 8.0, 16.0z); ¹³C-NMR (100 MHz, DMSO-d₆+D₂O): δ 156.7 (C-7), 156.5 (C-5), 155.7 (C-9), 145.2 (C-4’), 145.1 (C-3’), 130.8 (C-1’), 118.8 (C-6’), 115.6 (C-5’), 114.7 (C-2’), 107.3 (C-1’’), 102.1 (C-10), 96.0 (C-8), 95.3 (C-6), 81.4 (C-2), 78.9 (C-3’’), 76.3 (C-2’’), 74.3 (C-4’’), 66.3 (C-3), 62.4 (C-5’’), 27.8 (C-4).

Compound catechin 7-O-beta-D apiofuranoside was brown amorphous powder. The spot was detected under UV radiation at 254 nm, the black spot was detected by spraying with FeCl₃ and dark brown and orange spots was detected by spraying 10 % H₂SO₄ and anisaldehyde-H₂SO₄ followed by heating, respectively.

An aromatic ABX-spin system, the presence of meta-coupled aromatic signal [δ 6.74 (1H, d, J=2.0 Hz, H-2’)], ortho-coupled aromatic signal [δ 6.69 (1H, d, J=8.4 Hz, H-5’)] and ortho-meta-coupled aromatic signal [δ 6.59 (1H, dd, J=2.0, 8.4 Hz, H-6’)] was showed in ¹H-NMR spectrum. The two carbon signals of C-3’ and 4’ were observed in downfield (δ 145.1, 145.2) compared with the peaks of C-2’, 5’ and 6’ (δ 114.7, 115.6 and 118.8) in ¹³C-NMR spectrum. These meant B-ring was pyrocatechol moiety which was substituted by hydroxyl group in C-3’ and 4’. The signals of two more aromatic protons, which were meta-coupled ones [δ 5.90 (1H, d, J=2.4 Hz, H-6), δ 6.09 (1H, d, J=2.4 Hz, H-8)] were observed in ¹H-NMR spectrum, and the three carbon signals of C-5, C-7 and C-9, (δ 155.7, 156.5 and 156.7) were deshielded rather than C-6, 8 and 10 (δ 95.3, 96.0 and 102.1) in ¹³C-NMR spectrum.

These implied A-ring was related to phloroglucinol moiety which was substituted by hydroxyl group in C-5, 7 and 9. However the five carbon signals of 62.4 (api-5), 74.3 (api-4), 76.3 (api-2), 78.9 (api-3), 107.3 (api-1) indicate sugar existence in ¹³C-NMR spectrum. These signals were indicative of a D-apiofuranoside (Shengjun et al., 2000). The linkage of this sugar at C-7 was established by comparing the spectral data with the values reported in the literature (Na et al., 2002). Moreover, the proton signals of H-2 [δ 4.55 (1H, d, J=7.2 Hz)], H-3 [δ 3.89 (1H, m), H-4_eq [δ 2.65 (1H, dd, J=4.8, 16.0 Hz)] and H-4_ax [δ 2.40 (1H, dd, J=8.0, 16.0Hz)] were observed in ¹H-NMR and the carbon signals in ¹³C-NMR spectrum of C-2, 3 and 4 (δ 81.4, 66.3 and 27.8) indicated flavan-3ol moiety. Especially, the large coupling constant of H-2 (J=7.2 Hz) in ¹H-NMR and carbon signals of C-2 at δ 81.4 in ¹³C-NMR suggested a 2,3-trans-configuration.(Fig. 1, 2)

Fig. 1. Chemical structures of compound 1 isolated from Ulmus species.

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Fig. 2. HPLC chromatogram of compound 1. Sample 1 = Com. 1 1,000ppm(purple color line), Sample 2 = Com. 1 500 ppm (mint color line), Sample 3 = Com. 1 100 ppm (green color line), Sample 4 = Com. 1 50 ppm(blue color line).

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Thus, compound 1 was identified as catechin 7-O-β-D apiofuranoside based on the spectral data compared with the valuesreported in the previous studies (Jung et al., 2007; Bae et al., 2000; Shengjun et al., 2000; Na et al., 2002).

We quantified catechin 7-O-beta-D apiofuranoside from the stem and bark extracts of U. davidiana var. japonica for. Suberosa [172.35 ± 0.158 ppm (μg/mL), 17.24 ± 0.016%] (Fig. 3), the leaf extracts of U. davidiana Planchon [191.38 ± 0.219 ppm (μg/mL), 19.14 ± 0.022%] (Fig. 4), the stem and bark extracts of U. davidiana Planchon [177.84 ± 0.209 ppm (μg/mL), 17.78 ± 0.021%] (Fig. 5), the leaf extract of U. davidiana var. japonica [176.11 ± 0.155 ppm (μg/mL), 17.61 ± 0.015%] (Fig. 6), the stem extract of U. davidiana var. japonica [41.27 ± 0.191 ppm (μg/mL), 4.13 ± 0.019%] (Fig. 7), the stem and bark extracts of U. parvifolia Jacq [150.88 ± 0.205 ppm (μg/mL), 15.09 ± 0.020%] (Fig. 8) and the stem extract of U. pumila Linne [61.69 ± 0.178 ppm (μg/mL), 6.17 ± 0.018%] (Fig. 9), using a calibration equation (y = 6857.3x – 42331; R² = 0.9985). According to the results of the content analysis experiment using HPLC, there was no significant difference in the content of the indicator substance in the other samples except for the two samples (the stem extract of U. davidiana var. japonica and the stem extract of U. pumila Linne), but the two samples with a significantly lower level were important experimental results when evaluating the value of future use of raw materials. It is expected to be used as important basic data in future resource utilization.

Fig. 3. HPLC chromatogram of the stem and bark extracts of U. davidiana var. japonica for. suberosa. Sample 1 = stem and bark extracts of U. davidiana var. japonica for. suberosa. 1,000 ppm.

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Fig. 4. HPLC chromatogram of the leaf extract of U. davidiana Planchon. Sample 1 = leaf extract of U. davidiana Planchon.1,000 ppm.

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Fig. 5. HPLC chromatogram of the stem and bark extracts of U. davidiana Planchon. Sample 1 = stems and bark extract of U. davidiana Planchon.1,000 ppm.

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Fig. 6. HPLC chromatogram of the leaf extract of U. davidiana var. japonica. Sample 1 = leaf extract of U. davidiana var. japonica.1,000 ppm.

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Fig. 7. HPLC chromatogram of the stem extract of U. davidiana var. japonica. Sample 1 = stem extract of U. davidiana var. japonica.1,000 ppm.

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Fig. 8. HPLC chromatogram of the stem and bark extracts of U. parvifolia Jacq. Sample 1 = stems and bark extract of U. parvifolia Jacq.1,000 ppm.

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Fig. 9. HPLC chromatogram of the stem extract of U. pumila Linne. Sample 1 = stem extract of U. pumila Linne.1,000 ppm.

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4. CONCLUSION

This is the first report of the isolation and identification of catechin 7-O-beta-D apiofuranosidefrom U. davidiana var. japonica for. suberosa., U. parvifolia Jacq and U. davidiana Planchon.Since the initial isolation of catechin 7-O-beta-D apiofuranoside from U. Americana (Doskotch et al., 1973), it has been found to be distributed among U. davidiana var. japonica (Jung et al., 2007) and U. macrocarpa Hance (Kwon et al., 2011).

Based on the above results, it is expected that it will be used as an important research result in searching for new natural materials that can be given high added value.

ACKNOWLEDGMENT

This study was supported by the ‘R&D Program for Forest Science Technology (Project No. 201703 1B10-1819-BA01)’ offered by Korea Forest Service (Korea Forestry Promotion Institute), and this study was supported (in part) by 2020 Research Grant from Kangwon National University.

References

Bae Y.S., Kim J.K. Extractives of the bark of ash and elm as medicinal hardwood tree species. Journal of the Korean Wood Science and Technology. 2000; 28(3):62-69.

Cho S.K., Lee S.G., Kim C.J. Anti-inflammatory and analgesic activities of water extract of root bark of Ulmus parvifolia. Korean Journal of Pharmacognosy. 1996; 27(3):274-281.

Doskotch R.W., Milhail A.A., Chatterji S.K. Structure of the water-soluble feeding stimulant for Scolytus multistriatus: A revision. Phytochemistry. 1973; 12(5):1153-1155.

Hong N.D., Rho Y.S., Kim N.J., Kim J.S. A Study on efficacy of Ulmi cortex. Korean Journal of Pharmacognosy. 1990; 21(3):217-222.

Jun C.D., Pae H.O., Kim Y.C., Jeong S.J., Yoo J.C., Lee E.J., Choi B.M., Chae S.W., Park R.K., Chung H.T. Inhibition of nitric oxide synthesis by butanol fraction of the methanol extract of Ulmus davidiana in murine macrophages. Journal of Ethnopharmacology. 1998; 62(2):129-135.

Jung T.H. Korean Flora. 1974 Seoul: I-Mun Pubishing Co., Ltd. .

Jung M.J., Heo S.I., Wang M.H. Reducing power and α-glucosidase inhibitory profiles of (-)-catechin and its glycoside. Korean Journal of Pharmacognosy. 2007; 38(4):358-362.

Kim S.H., Hwang K.T., Park J.C. Isolation of flavonoids and determination of rutin from the leaves of Ulmus parvifolia. Korean Journal of Pharmacognosy. 1992; 23(4):229-234.

Kim J.P., Kim W.G., Koshino H., Jung J., Yoo I.D. Sesquiterpene O-naphthoquinones from the root bark of Ulmus davidiana. Phytochemistry. 1996; 43(2):425-430.

10.

Kim C.S., Lee J.M., Choi C.O., Park S.B., Eom T.J. Chemical analysis and isolation of antibacterial compound from Ulmus species (1): chemical analysis and antibacterial activity of extractives. Journal of the Korean Wood Science and Technology. 2002; 30(4):66-73.

11.

Kwon J.H., Kim S.B., Park K.H., Lee M.W. Antioxidative and anti-inflammatory effects of phenolic compounds from the roots of Ulmus macrocarpa. Archives of Pharmacal Research. 2011; 34(9):1459-1466.

12.

Lee Y.N. Flora of Korea. 1996 Seoul: Kyo-Hak Pubishing Co., Ltd. .

13.

Lee Y.J., Han J.P. Antioxidant activities and nitrite scavenging abilities of extracts from Ulmus devidianan. Journal of the Korean Society of Food Science and Nutrition. 2000; 29(5):893-899.

14.

Lee G.H., Shim C.J., Chang Y.I., Seong H., Oh H.R., Oh M.J. Research Papers: Antimicrobial activities of (-) epicatechin from Ulmus davidiana var. japonica cortex. Journal of Food Science and Nutrition. 2001; 6(4):230-234.

15.

Moon Y.H., Rim G.R. Studies on the constituents of Ulmus parvifolia. Korean Journal of Pharmacognosy. 1995; 26(1):1-7.

16.

Na M.K., An R.B., Lee S.M., Min B.S., Kim Y.H., Bae K.H., Kang S.S. Antioxidant compounds from the stem bark of Sorbus commixta. Natural Product Sciences. 2002; 8(1):26-29.

17.

Park S.H., Goo Y.M., Na D.S. Isolation and structure elucidation of a catechin glycoside with phospholipase A2 inhibiting activity from Ulmi cortex. Bulletin of the Korean Chemical Society. 1996; 17(2):101-103.

18.

Shengjun G., Elisabet F., Lennart K., Bengt R. Triterpenoid saponins containing an acetylated branched D-fucosyl residue from Quillaja saponaria Molina. Phytochemistry. 2000; 53(8):861-868.

19.

Son B.W., Park J.H., Zee O.P. Catechin glycoside from Ulmus davidiana. Archives of Pharmacal Research. 1989; 12(3):219-222.

20.

Yang Y., Hyun J.W., Lim K.H., Kim H.J., Woo E.R., Park J. Antineoplastic effect of extracts from traditional medicinal plants and various plants (III). Korean Journal of Pharmacognosy. 1996; 27(2):105-110.