Chemical constituents from the mycelium of isaria japonica yasuda - Nguyen Ngoc Tuan

Journal of Science and Technology 54 (2C) (2016) 334-340 CHEMICAL CONSTITUENTS FROM THE MYCELIUM OF ISARIA JAPONICA YASUDA Nguyen Ngoc Tuan, Nguyen Tan Thanh, Dao Thi Thanh Xuan, Tran Dinh Thang * Department of Chemistry, Vinh University, 182 Le Duan, Vinh City, Nghe An province, Vietnam *Email: thangtd@vinhuni.edu.vn Received: 15 June 2016; Accepted for publication: 23 October 2016 ABSTRACT A chemical investigation of the mycelium of Isaria japnonica Yasuda resulted in the identification of eight compounds, including two steroids (ergosterol, ergosterol peroxide), one flavonoid (tricine), one phenolic (2-hydroxy-3-phenylpropanoic acid), two nitrogenous compounds (adenosine, uracil), one saccharide (D-mannitol), one amino acid (3-amino butanoic acid). The chemical structures of nine compounds were determined on the basis of 1D and 2D NMR, UV, IR and MS analytical results. Keywords: Isaria japnonica, triterpenoid, steroid, flavonoid, phenolic. 1. INTRODUCTION Natural remedies are becoming increasingly popular and important in the public and scientific communities. Historically, natural remedies have been shown to present interesting biological and pharmacological activity and are used as chemotherapeutic agents. For centuries Cordyceps is a genus of more than 400 species in the family Clavicipitaceae. All Cordyceps species are endoparasitoids, parasitic mainly on insects and other arthropods, a few are parasitic on other fungi. Until recently, the best known species of the genus was Cordyceps sinensis [1, 2, 3]. Isaria japonica Yasuda has traditionally been used as health foods for various diseases in Japan, Korea and China [2, 3, 5]. Myriocin, a sphingosine analog isolated from the culture filtrate of Isaria sinclairii (P. cicadae), showed inhibitory effect on T cell-dependent immune responses [5]. Recently, they reported about the success in cultivating I. japonica (P. tenuipes) in a liquid medium and demonstrated that this liquid medium augmented anti-sheep red blood cell IgM plaque-forming cells response upon oral administration in mice [5, 6]. In the present study, we have succeeded in cultivating Isaria japnonica in a liquid medium and chemical investigation of the mycelium of I. japnonica resulted in the identification of eight compounds including two steroids (ergosterol, ergosterol peroxide), one flavonoid (tricine), one phenolic (2-hydroxy-3-phenylpropanoic acid), two nitrogenous compounds (adenosine, uracil), one saccharide (D-mannitol), one amino acid (3-amino butanoic acid). The chemical structures of the eight compounds were determined on the basis of 1D and 2D NMR, UV, IR and MS analytical results. Chemical constituents from the mycelium of Isaria Japonica Yasuda 335 2. EXPERIMENTS 2.1. General Melting points were determined using Yanagimoto MP-S3 apparatus. Optical rotations were measured using a JASCO DIP-370 polarimeter. The UV spectra were obtained on a Hitachi UV-3210 spectrophotometer, and IR spectra were recorded on a Shimadzu FTIR-8501 spectrophotometer. 1H- and 13C-NMR, COSY, NOESY, HMQC, and HMBC spectra were obtained on the Bruker AV-500 NMR spectrometer, with tetramethylsilane (TMS) as the internal standard and chemical shifts were reported in δ values (ppm). The electrospray ionization (ESI) mass spectra were determined using an Agilent 1200 LC-MSD Trap spectrometer. Column chromatography (CC) was performed on silica gel (Kieselgel 60, 70-230 mesh and 230-400 mesh, E. Merck). Thin layer chromatography (TLC) was conducted on precoated Kieselgel 60 F 254 plates (Merck) and the compounds were visualized by spraying with 10 % (v/v) H2SO4 followed by heating at 110oC for 10 min. 2.2. Fungal material The entomogenous fungi I. japonica Yasuda was collected at the Pumat National Park of Nghean Province, Vietnam, in November 2013 and identified by Assoc. Prof. Dr. Tran Ngoc Lan, Institute for Regional Research and Development, Ministry of Science and Technology, Vietnam. A voucher specimen (VU130805) was deposited at the herbarium of the Department of Chemistry, Vinh University. I. japonica was cultured in a Potato Dextrose Agar (PDA) (5 L), at 25 – 26 °C for 14 days. 2.3. Extraction and isolation The cultures were filtered through cheese cloth to separate broth and mycelium. The mycelium of I. japonica was extracted with methanol (1 L × 3) at ambient temperature, and the combined extracts were concentrated under reduced pressure to give a deep brown syrup (25 g). The crude extract was suspended in water and partitioned with ethyl acetate to afford ethyl acetate (15 g) and water soluble (10 g) fractions, respectively. The ethyl acetate soluble extracts were applied to silica gel column chromatography with a mixture of hexane and acetone step gradient system (100:0, 25:1, 15:1, 10:1, 7:1, 5:1) and then eluted with a chloroform:methanol step gradient solvent mixture (10:0, 6:1, 3:1, 2:1, 1:1) to afford minor fractions. These fractions were monitored by TLC to combine into six fractions. Fraction 1 was subjected to the silica gel column chromatography (200 g, 60 × 3 cm) eluting with a hexan:axeton (15:1) to afford compound 1 (123 mg) and compound 3 (15 mg). Fraction 4 was subjected to the silica gel column chromatography (200 g, 60×3 cm) eluting with a hexan:axeton (9:1) to afford compound 2 (31 mg). Fraction 3 (1.2 g) was subjected to the silica gel column chromatography (200 g, 60 × 3 cm) eluting with a chloroform/methanol step gradient system (15:1, 9:1, each 200 mL) to afford compound 4 (13 mg) and compound 5 (25 mg). Fraction 3 was subjected to the silica gel column chromatography (200 g, 60×3 cm) eluting with a chloroform:metanol (30:1) to afford compound 5 (38 mg). Fraction 5 afforded compound 6 (30 mg), compound 7 (16.5 mg), and compound 8 (14.5 mg). Compound 1: white powder, m.p. 165 – 167 oC; UV (MeOH) λmax nm: 211, 285. IR (KBr) νmax (cm-1): 3433, 2959, 1726, 1090; EI-MS m/z 396 [M]+; 1H-NMR (500 MHz, CDCl3) (δ ppm): 5.49 (1H, m), 5.35 (1H, m), 5.28 (1H, dd, J = 15.5, 7.5 Hz), 5.25 (1H, dd, J = 15.5, 7.0 Hz), 3.48 Nguyen Ngoc Tuan, Nguyen Tan Thanh, Dao Thi Thanh Xuan, Tran Dinh Thang 336 (1H, m), 1.05 (3H, d, J = 7.0 Hz), 0.96 (3H, s), 0.93 (3H, d, J = 7.0 Hz), 0.85 (3H, d, J = 6.5 Hz), 0.82 (3H, d, J = 6.5 Hz), 0.61 (3H, s), 13C-NMR (125 MHz, CDCl3) (δ ppm): 140.3 (C-5), 139.8 (C-8), 135.0 (C-23), 131.2 (C-22), 118.2 (C-6), 115.8 (C-7), 68.3 (C-3), 55.0 (C-17), 53.6 (C- 14), 45.5 (C-9), 42.2 (C-13), 41.8 (C-24), 40.4 (C-4), 40.0 (C-20), 39.0 (C-1), 38.2 (C-12), 37.7 (C-10), 32.2 (C-25), 31.5 (C-2), 27.4 (C-16), 22.2 (C-15), 20.6 (C-11), 20.4 (C-27), 19.4 (C-26), 19.1 (C-21), 17.0 (C-28), 15.8 (C-19), 11.5 (C-18). Compound 2: Colorless needles, m.p. 176 - 178 0C; EI-MS m/z 428 [M]+; 1H-NMR (500 MHz, CDCl3) (δ ppm): 6.44 (1H, d, J=8.5 Hz, H-7), 6.23 (1H, d, J = 8.5 Hz, H-6), 5.28 (1H, m, H-22), 5.18 (1H, m, H-23), 3.58 (1H, m, H-3), 1.05 (3H, d, J = 6.5 Hz, H-21), 0.96 (3H, d, J = 7.0 Hz, H-28), 0.95 (3H, s, H-19), 0.89 (3H, d, J = 6.5 Hz, H-27), 0.87 (3H, s, H-18), 0.87 (3H, d, J = 6.5 Hz, H-26); 13C-NMR (125 MHz, CDCl3) (δ ppm): 135.6 (C-22), 135.2 (C-6), 131.5 (C-23), 130.1 (C-7), 81.4 (C-5), 78.4 (C-8), 64.6 (C-3), 55.4 (C-17), 51.2 (C-14), 50.9 (C-9), 44.0 (C-13), 42.0 (C-24), 40.1 (C-20), 38.7 (C-4), 36.9 (C-10), 36.5 (C-12), 34.5 (C-2), 32.4 (C- 25), 29.9 (C-1), 28.2 (C-16), 22.8 (C-15), 21.7 (C-11), 20.2 (C-21), 19.7 (C-27), 19.4 (C-26), 17.9 (C-19), 17.2 (C-28), 12.5 (C-18). Compound 3: Pale yellow powder, m.p. 296 – 298 0C; ESI-MS m/z: 331 [M+H]+; 1H-NMR (500 MHz, DMSO-d6) (δ ppm): 7.32 (2H, s, H-2 and H-6), 6.97 (1H, s, H-3), 6.55 (1H, d, J = 2.0 Hz, H-8), 6.20 (1H, d, J = 2.0 Hz, H-6), 3.88 (6H, s, 3’-OCH3 and 5’-OCH3) ; 13C-NMR (125 MHz, DMSO-d6) (δ ppm): 181.8 (C-4), 164.1 (C-7),163.6 (C-2), 161.4 (C-9), 157.3 (C-5), 148.2 (C-3′ and 5′), 139.9 (C-4′), 120.4 (C-1′), 104.4 (C-2′ and 6′), 103.7 (C-10), 103.6 (C-3), 98.8 (C-6), 94.2 (C-8), 56.4 (2C, 3’-OCH3 and 5’-OCH3). Compound 4: White powder, [α]D24 −20.8(c 2, H2O) ; m.p. 122 – 124 °C ; 1H-NMR (CDCl3&MeOD, 500 MHz) (δ ppm): 7.35 (2H, m, H-3’,5’), 7.28 (1H, t, J = 7.5 Hz, H-4’), 7.11 (2H, brd, J = 7.0 Hz, H-2’,6’), 4.10 (1H, dd, J = 6.5, 4.0 Hz, H-2), 2.94 (1H, dd, J = 7.0, 3.5 Hz, H-3a), 2.17 (1H, dd, J = 8.0, 7.5 Hz, H-3b); 13C-NMR (CDCl3&MeOD, 125 MHz) (δ ppm): 167.3 (C-1), 135.6(C-1’), 130.2(C-2’,6’), 129.1(C-3’,5’), 127.6(C-4’), 56.5(C-2), 40.3(C-3). Compound 5: Colorless powder; [α]D25 -31.7 (c 0.04, MeOH); m.p. 235 - 237 ℃; EI-MS (rel. int.): m/z 267 ([M] +, 3), 178(30), 164(78), 135(100) ; UV (MeOH) λmax: 261 nm; IR (neat) νmax: 3334, 1654 cm-1; 1H-NMR (CDCl3 & CD3OD, 500 MHz) (δ ppm): 3.77 (1H, dd, J = 12.4, 2.4 Hz, H-5′), 3.97 (1H, dd, J = 12.4, 2.4 Hz, H-5′), 4.27 (1H, q, J = 2.4 Hz, H-4′), 4.35 (1H, dd, J = 4.8, 2.4 Hz, H-3′), 4.79 (1H, dd, J = 7.2, 4.8 Hz, H-2′), 5.86 (1H, d, J = 7.2 Hz, H-1′), 8.05 (1H, s, H-8), 8.23 (1H, s, H-2); 13C-NMR (CDCl3 & CD3OD, 125 MHz) (δ ppm): 156.2 (C- 6), 152.5 (C-2), 148.6 (C-4), 141.0 (C-8), 120.6 (C-5), 91.3 (C-1’), 87.6 (C-4’), 74.2 (C- 2’), 71.9 (C-3’), 61.6 (C-5’). Compound 6: Light brown powder; m.p. 310 – 311 0C ; EI-MS (rel. int.): m/z 112 ([M]+, 100), 69(52); UV (MeOH) λmax: 258 nm; IR (neat) νmax: 3116, 2926, 2856, 1711, 1671 cm-1; 1H- NMR (DMSO-d6 , 500 MHz) (δ ppm): 5.44 (1H, d, J = 7.5 Hz, H-5), 7.37 (1H, d, J = 7.5 Hz, H- 6), 10.81 (1H, br s), 10.98 (1H, br s). Compound 7: Colorless powder; m.p. 166-1680C, EI-MS (rel. int.): m/z 182 ([M]+, 3), 103(60), 73(100); IR (neat) νmax: 3207, 2918, 1420, 1380 cm-1; 1H-NMR (DMSO-d6 , 500 MHz) (δ ppm): 4.40 (1H, d, J=5.0Hz, 2-OH), 4.31 (1H, t, J = 5.5 Hz, 1-OH), 4.12 (1H, d, J = 7.0 Hz, 3-OH), 3.35-3.63 (4H, m, H-1,2,3); 13C-NMR (DMSO-d6 , 125 MHz) (δ ppm): 64.1 (C-1), 69.9 (C-3), 71.5 (C-2). Chemical constituents from the mycelium of Isaria Japonica Yasuda 337 Compound 8: Colorless powder; m.p. 189 – 190 ℃ ; 1H-NMR (CDCl3, 500 MHz) (δ ppm): 5.25 (1H, dt, J = 6.5, 13.0 Hz, H-3), 2.60 (1H, dd, J = 7.0, 7.0 Hz, H-2), 2.47 (1H, dd, J = 7.0, 7.0 Hz, H-2), 1.27 (3H, d, J = 6.5 Hz, H-4); 13C-NMR (DMSO-d6 , 125 MHz) (δ ppm): 169.2 (C-1), 67.6 (C-3), 40.8 (C-2), 19.8 (C-4). 3. RESULTS AND DISCUSSION Compound 1 was obtained as an optically active white powder. The EI-MS of 1 showed a pseudomolecular ion peak at m/z 396 [M]+, corresponding to a molecular formula of C28H44O. The UV spectrum of 1 exhibited absorption maxima at 211 nm and 285 nm. The IR absorption bands at 3433, 2959, 1726, 1090 cm-1 suggested the presence of hydroxy group, carbon−carbon double bond functionalities, respectively. The 1H- NMR spectrum of 1 displayed six methyl groups [δH 0,61 (3H, s), 0,82 (3H, d, J = 6,5 Hz), 0,85 (3H, d, J = 6,5 Hz), 0,93 (3H, d, J = 7,0 Hz), 0,96 (3H, s), and 1,05 (3H, d, J = 7,0 Hz)], respectively. In addition, the presence of one proton attached to an oxygen bearing carbon [δH 3,48 (1H, m)] and four protons attached to double bonds at [δH 5,28 (1H, dd, J = 15,5, 7,5 Hz), 5,25 (1H, dd, J = 15,5, 7,0 Hz), 5,35 (1H, m), and 5,49 (1H, m)]. Its 13C-NMR spectrum displayed totally 28 carbon signals, which confirmed the presences of six olefinic carbons [δC 116,3; 119,6; 130,0; 135,6; 140,7 and 142,0] and one oxygenated carbon [δC 70,5 ppm]. These facts indicated that compound 1 is ergosterol, confirmed by direct comparison with the literature data [7]. Compound 2 was obtained as an optically active colorless needles with optical rotation .... The EI-MS of 2 showed a pseudomolecular ion peak at m/z 428 [M]+, corresponding to a molecular formula of C28H44O3. The 1H- NMR spectrum of 2 displayed six methyl at [δH 0,87 (s, H- 18), 0,95 (s, H-19), 0,87 (d, J=6,5 Hz, H-26), 0,89 (d, J=6,5 Hz, H-27), 0,96 (d, J=7,0 Hz, H- 28), 1,05 (d, J=6,5 Hz, H-21)], four protons attached to double bonds at [δH 6,23 (J=8,5 Hz, H- 6), 6,44 d (J=8,5 Hz, H-7), 5,28 (m, H-22)] and 5,18 (m, H-23)], and one proton attached to an oxygen bearing carbon at [δH 3,58 (m, H-3)], which suggested the presence of the sterol. Its 13C- NMR and DEPT spectrum displayed totally 28 carbon signals, which confirmed the presences of six methyl carbons, seven methylen, eleven methin and four quaternary carbons. In addition, the 13C-NMR spectrum of compound 2 exhibited signals consistent with the presence of three oxygenated carbons [81,4 (C-5); 78,4 (C-8); 64,6 (C-3)] and two C=C double bonds in the downfield region [135,6 (C-22) and 131,5 (C-23); 135,2 (C-6) and 130,1 (C-7)], which were similar to those observed for compound 1, except for the absence of one C=C double bond. Notably, these spectroscopic data were consistent with those reported in the literature for a known compound ergosterol peroxide (5α, 8-epidioxyergosta-6,22-dien-3-ol) [7] and compound 2 was therefore characterized as deduced. Compound 3 was obtained as an optically active pale yellow powder. The ESI-MS showed a pseudomolecular ion peak at m/z 331 [M+H]+, corresponding to a molecular formula of C17H14O7. The 1H-NMR spectrum showed signals due to five sp2-methine protons [δH 7.32 (2H, s, H-2 and H-6), 6.97 (1H, s, H-3), 6.55 (1H, d, J = 2.0 Hz, H-8), 6.20 (1H, d, J = 2.0 Hz, H-6)], and two methoxy protons [δH 3.88 (6H, s, 3’-OCH3 and 5’-OCH3)]. The 13C-NMR showed seventeen carbon signals at 181.8 (C-4), 164.1 (C-7),163.6 (C-2), 161.4 (C-9), 157.3 (C-5), 148.2 (2C, C-3′ and 5′), 139.9 (C-4′), 120.4 (C-1′), 104.4 (2C, C-2′ and 6′), 103.7 (C-10), 103.6 (C-3), 98.8 (C-6), 94.2 (C-8), 56.4 (2C, 3’-OCH3 and 5’-OCH3). Compounds 3 was identified as Nguyen Ngoc Tuan, Nguyen Tan Thanh, Dao Thi Thanh Xuan, Tran Dinh Thang 338 tricin by comparison of its physical and spectroscopic properties with those reported in the literature [8, 9]. Compound 4 was obtained as an optically active white powder, m.p. 122 – 124 °C. The 1H- NMR spectrum of 4 displayed the signals of the ABX system of a mono-substituted benzene ring [δH 7.35 (2H, m, H-3’,5’), 7.28 (1H, t, J = 7.5 Hz, H-4’), 7.11 (2H, brd, J = 7.0 Hz, H- 2’,6’)], one oxygenated methine proton [δH 4.10 (1H, dd, J = 6.5, 4.0 Hz, H-2)], two methylen protons [δH 2.94 (1H, dd, J = 7.0, 3.5 Hz, H-3a), 2.17 (1H, dd, J = 8.0, 7.5 Hz, H-3b)]. The 13C- NMR spectrum of 4 showed one carbonyl carbon [δC 67.3 (C-1)], six aromatic carbons [δC 135.6(C-1’), 130.2(C-2’,6’), 129.1(C-3’,5’), 127.6(C-4’)], one oxygenated carbon [δC 56.5(C- 2)], one methylene carbon [δH 40.3(C-3)]. Compounds 3 was identified as 2-hydroxyl-3- phenylpropanoic acid by comparison of its physical and spectroscopic properties with those reported in the literature [10]. Compound 5 was obtained as an optically active colorless powder. The IR absorption bands at 3334, 1654 cm-1. The UV spectrum of 5 exhibited absorption maxima at 261 nm. The EI-MS of 5 showed a pseudomolecular ion peak at m/z 267, corresponding to a molecular formula of C4H4N2O2. The 1H-NMR spectrum of 5 displayed the signals of a ribose sugar moiety at 3.77 (1H, dd, J = 12.4, 2.4 Hz, H-5′), 3.97 (1H, dd, J = 12.4, 2.4 Hz, H-5′), 4.27 (1H, ddd, J = 2.4, 2.4, 2.4 Hz, H-4′), 4.35 (1H, dd, J = 4.8, 2.4 Hz, H-3′), 4.79 (1H, dd, J = 7.2, 4.8 Hz, H-2′), 5.86 (1H, d, J = 7.2 Hz, H-1′), and adenine moiety at 8.05 (1H, s, H-8), 8.23 (1H, s, H-2). The 13C-NMR spectrum of 5 also displayed the signals of a ribose sugar moiety at 91.3 (C- 1’), 87.6 (C-4’), 74.2 (C-2’), 71.9 (C-3’), 61.6 (C-5’), and adenine moiety at 156.2 (C- 6), 152.5 (C-2), 148.6 (C-4), 141.0 (C-8), 120.6 (C-5). The HMBC spectrum of 5 suggested adenine attached to a ribose sugar molecule (ribofuranose) moiety via a β-N9- glycosidic bond. Compounds 5 was identified as adenosine by comparison of its physical and spectroscopic properties with those reported in the literature [11]. Compound 6 was obtained as an optically active light brown powder, m.p. 310 – 311 0C. The IR absorption bands at 3116, 2926, 2856, 1711, 1671 cm-1. The UV spectrum of 6 exhibited absorption maxima at 258 nm. The EI-MS of 6 showed a pseudomolecular ion peak at m/z 112, corresponding to a molecular formula of C4H4N2O2. The 1H-NMR spectrum showed signals due to two sp2-methine protons [δH 5.44 (1H, d, J = 7.5 Hz, H-5), 7.37 (1H, d, J = 7.5 Hz, H-6)], two proton NH at 10.81 (1H, br s), 10.98 (1H, br s). Compounds 6 was identified as uracil by comparison of its physical and spectroscopic properties with those reported in the literature [12]. Compound 7 was obtained as an optically active colorless powder, m.p. 166 - 168 ℃. The IR absorption bands at 3207 (OH), 2918, 1420, 1380 cm-1. The EI-MS of 7 showed a pseudomolecular ion peak at m/z 182 corresponding to a molecular formula of C6H14O6. The 1H- NMR spectrum of 7 displayed the signals of four hydroxyl groups [δH 4.40 (1H, d, J = 5.0Hz, 2- OH), 4.31 (1H, t, J = 5.5 Hz, 1-OH), 4.12 (1H, d, J = 7.0 Hz, 3-OH)], and four methine protons at 3.35-3.63 (4H, m, H-1,2,3). The 13C-NMR spectrum of 7 showed three oxygenated carbons 64.1 (C-1), 69.9 (C-3), 71.5 (C-2). Compounds 7 was identified as D-mannitol by comparison of its physical and spectroscopic properties with those reported in the literature [13]. Compound 8 was obtained as an optically active colorless powder, m.p. 189 – 190 ℃. The 1H-NMR spectrum of 8 displayed totally seven protons signals, which confirmed the presences of one proton attached to an amino bearing carbon at 5.25 (1H, dt, J = 6.5 Hz, H-3), methylen protons at 2.60 (1H, dd, J = 7.0, 7.0 Hz, H-2) and 2.47 (1H, dd, J = 7.0, 7.0 Hz, H-2), methyl proton at 1.27 (3H, d, J = 6.5 Hz, H-4). In addition, the 13C-NMR and DEPT spectrums Chemical constituents from the mycelium of Isaria Japonica Yasuda 339 displayed totally four carbons signals, including one carbonyl carbon at 69.2 (C-1), one nitrogenated carbon 67.6 (C-3), one methylen carbon at 40.8 (C-2) and one methyl carbon 19.8 (C-4). Compounds 8 was identified as 3-amino butanoic acid by comparison of its physical and spectroscopic properties with those reported in the literature [14]. NH NH O O Ergosterol (1) Ergosterol peroxide (2) Tricine (3) Uracil (6) 2-hydroxyl-3-phenylpropanoic acid (4) Adenosine (5) D-mannitol (7) 3-amino butanoic acid (8) 4. CONCLUSION In the present study, we have succeeded in cultivating I. japnonica in a liquid medium and studying chemical composition of the mycelium of I. japnonica Yasuda in Pumat, Nghean province resulted in the identification of eight compounds, including two steroids (ergosterol, ergosterol peroxide), one flavonoid (tricine), one phenolic (2-hydroxy-3-phenylpropanoic acid), two nitrogenous compounds (adenosine, uracil), one saccharide (D-mannitol), one amino acid (3-amino butanoic acid). The chemical structures of the eight compounds were determined on the basis of 1D and 2D NMR, UV, IR and MS analytical results. REFERENCES 1. John H. and Matt C. - Medicinal value of the caterpillar fungi species of the genus Cordyceps (Fr.) Link (Ascomycetes): A Review, Int. J. Med. Mushrooms 10 (3) (2008) 219-234. 2. Zhu J. S., Halpern G. M., and Jones K. - The scientific rediscovery of an ancient Chinese herbal medicine: Cordyceps sinensis: Part I, J. Altern. Complement Med. 4 (1998) 289- 303. 3. Zhu J. S., Halpern G. M., and Jones K. - The scientific rediscovery of a precious ancient Chinese herbal regimen: Cordyceps sinensis: Part II, J. Altern. Complement Med. 4 (1998) 429- 457. 4. Takano F., Kikuchi Y., Fushiya S., Hojo N., Nozoe S., and Yahagi N. - The culture fluid of Isaria japonica Yasuda augments anti-sheep red blood cell antibody response in mice, Biol. Pharm. Bull. 19 (1996) 641-643. 5. Shin K. H., Lim S. S., Lee S., Lee Y. S., Jung S. H. and Cho S. Y. - Antitumour and immuno-stimulating activities of the fruiting bodies of Paecilomyces japonica, a new type of Cordyceps spp., Phytother Res. 17 (2003) 830 – 833. Nguyen Ngoc Tuan, Nguyen Tan Thanh, Dao Thi Thanh Xuan, Tran Dinh Thang 340 6. Yahagi N., Komatsu M., Hiramatsu M., Shi H., Yahagi R., and Kobayashi H. - Radical scavenging activities of condensed culture medium of Isaria japonica Yasuda and hot water extract of Fomes fomentarius (L.: Fr) Kicrx., Nat. Med. J. 53 (1999) 319-323. 7. Yoshihisa T., Minoru U., Takashi O., Kimiko N., Koutarou M., and Toshiaki T. - Glycosides of ergosterol derivatives from Hericum erinacens, Phytochemistry 30 (12) (1991) 4117-4120. 8. Stochmal A., Simonet A. M., Macias F. A., and Oleszek W. J. - Alfalfa (Medicago sativa L.) flavonoids. 2. Tricin and chrysoeriol glycosides from Aerial Parts, Agric. Food Chem. 49 (2001) 5310-5314. 9. Kuwabara H., Mouri K., Otsuka H., Kasai R., and Yamasaki K. - Tricin from a Malagasy Connaraceous Plant with potent antihistaminic activity, J. Nat. Prod. 66 (2003) 1273- 1275. 10. Evidente A., Iacobellis N.S., Scopa A., and Surico G. - Isolation of β-phenyllactic acid related compounds from Pseudomonas syringae, Phytochemistry 29 (5) (1990) 1491-1497 11. Morton I. K. and Judith M. H. - Concise dictionary of pharmacological agents: Properties and Synonyms, Springer Science & Business Media, 2012, pp. 106. 12. Hurd R. E. and Reid B. R. - NMR spectroscopy of the ring nitrogen protons of uracil and substituted uracils; relevance to Aψ base pairing in the solution structure of transfer RNA, Nucleic Acids Res. Aug. 4 (8) (1977) 2747-2755. 13. Lee E. J., Lee J. Y., Kim J. S., and Kang S. S. - Phytochemical studies on Lonicerae Flos (1) – isolation of iridoid glycosides, Nat. Prod. Sci. 16 (1) (2010) 32-38 14. Weiß M., Brinkmann T., Gröger H. - Towards a greener synthesis of (S)-3-aminobutanoic acid: Process development and environmental assessment, Green Chemistry 12 (9) (2010) 1580-1588. TÓM TẮT THÀNH PHẦN HÓA HỌC CỦA LOÀI NẤM KÍ SINH CÔN TRÙNG (Isaria japonica) Ở VIỆT NAM Nguyễn Ngọc Tuấn, Nguyễn Tân Thành, Đào Thị Thanh Xuân, Trần Đình Thắng* Khoa Hóa, Trường Đại học Vinh, 182 Lê Duẩn, thành phố Vinh, tỉnh Nghệ An, Việt Nam *Email: thangtd@vinhuni.edu.vn Nghiên cứu thành phần hóa học của nấm kí sinh côn trùng (Isaria japnonica) bằng các phương pháp sắc kí đã phân lập được 8 hợp chất bao gồm 2 steroit (ergosterol, ergosterol peroxit), 1 flavonoit (tricin), 1 phenolic (axit 2-hydroxyl-3-phenyl-propanoic), 2 hợp chất chứa nitơ (adenosin, uracil), 1 saccarit (D-mannitol), 1 amino axit (axit 3-amino butanoic). Các hợp chất này được xác định cấu trúc bằng các phương pháp phổ tử ngoại (UV), phổ hồng ngoại (IR), phổ khối lượng phân giải cao (HR-ESI-MS), phổ cộng hưởng từ (1H-, 13C-NMR, DEPT, HMBC HSQC và COSY). Từ khóa: Isaria japnonica, triterpenoit, steroit, flavonoit, phenolic.