海洋紅樹林植物黃槿內(nèi)生真菌Aspergillus sydowii EN-198化學成分研究

杜豐玉, 李曉明, 李春順, 王斌貴

(1.中國科學院 海洋研究所 實驗海洋生物學重點實驗室, 山東 青島 266071; 2.中國科學院研究生院,北京100049)

紅樹植物一般分布于熱帶、亞熱帶潮間帶或河口邊緣, 生長環(huán)境特殊, 具有水陸兩棲特性, 蘊藏著豐富并極具特色的微生物資源[1]。紅樹林內(nèi)生真菌是指那些在其生活史中的全部或某一段時期生活在宿主體內(nèi), 但不引起宿主明顯病害癥狀的真菌。因為紅樹植物獨特的兩棲特性, 這些內(nèi)生真菌具有海洋微生物的特點, 擁有獨特的代謝途徑和遺傳背景, 其代謝產(chǎn)物化學多樣性也因而比較豐富。近年來紅樹林內(nèi)生真菌代謝產(chǎn)物研究已經(jīng)受到越來越多的關(guān)注[2-4]。

對于紅樹植物及其內(nèi)生真菌次級代謝產(chǎn)物的研究本研究組曾有報道[5-9]。本文報道從采自海南東寨港的海洋紅樹林植物黃槿(Hibiscus tiliaceus)葉中分離到一株內(nèi)生真菌Aspergillus sydowiiEN-198, 并從其發(fā)酵培養(yǎng)物中分離鑒定了14個化合物, 其中化合物1和2為首次從Aspergillus sydowii中分離得到。

1 材料與方法

1.1 儀器與試劑

Bruker Avance 500 MHz核磁共振儀, TMS內(nèi)標;薄層色譜硅膠 GF254和柱色譜硅膠(200～300目)為青島海洋化工廠分廠產(chǎn)品; Lobar LiChroprep RP-18硅膠 (40～63 μm, Merck); 顯色劑為茴香醛硫酸溶液和碘; 所用有機溶劑為重蒸的工業(yè)級溶劑。

1.2 菌株發(fā)酵

(1)菌株: 菌株 EN-198是分離自海南東寨港采集的海洋紅樹林植物黃槿(Hibiscus tiliaceus)葉中的內(nèi)生真菌。

(2)菌株發(fā)酵: 菌種以瓊脂–麥芽膏培養(yǎng)基, 4℃保存。發(fā)酵培養(yǎng)基組成為: 酵母膏3 g/L, 麥芽糖20 g/L, 山梨醇 20 g/L, 味精 10 g/L, 色氨酸 0.5 g/L,KH2PO40.5 g/L, MgSO4·7H2O 0.3 g/L, pH 6.5。250 mL三角燒瓶, 每瓶裝液體培養(yǎng)基150 mL, 115 ℃滅菌20 min后接種。共接種液體培養(yǎng)基15 L, 28 ℃, 160 r/min搖床培養(yǎng)7 d, 過濾, 分別收集菌絲體和發(fā)酵液。

圖1 化合物1–14的結(jié)構(gòu)Fig. 1 Structures of compounds 1–14

1.3 提取分離

收集發(fā)酵液約15 L, 用乙酸乙酯萃取。菌絲體涼干、粉碎后用丙酮∶水(4∶1)浸泡。將丙酮蒸出, 水相用乙酸乙酯萃取, 萃取物經(jīng)薄層層析檢測發(fā)現(xiàn)與發(fā)酵液萃取物基本一致, 合并得到粗提物10g。

將上述粗提物進行硅膠VLC柱層析, 以石油醚-乙酸乙酯、氯仿-甲醇體系梯度洗脫, 經(jīng)TLC檢測合并為10個組分(Fr.1-10), 其中組分Fr.4-5經(jīng)正相硅膠柱層析, Sephadex LH-20 (氯仿: 甲醇=1: 1)凝膠柱層析與反相硅膠柱層析分離得到化合物 11 (10.5 mg),12 (29.5 mg)與13 (24.1 mg); 組分Fr.7經(jīng)正相硅膠柱層析, Sephadex LH-20 (甲醇)凝膠柱層析和反相硅膠柱層析得到化合物1 (3.2 mg), 2 (4.1 mg), 3 (2.7 mg),4 (14.9 mg), 5 (41.2 mg), 6 (4.8 mg)與 7 (6.7 mg); 組分Fr.8-10經(jīng)正相硅膠柱層析, Sephadex LH-20 (甲醇)凝膠柱層析和制備薄層層析得到化合物8 (3.3 mg), 9(4.1 mg), 10 (5.2 mg)與 14 (21.2 mg)。

1.4 抑菌活性篩選

(1)病原指示菌: 金黃色葡萄球菌(Staphyloccocus aureus)與大腸桿菌(Escherichiacoli)。

(2)指示菌菌懸液制備: 指示菌接種于LB培養(yǎng)基表面, 于37 ℃培養(yǎng)24 h后, 吸取2 mL無菌0.85%NaCl溶液洗滌培養(yǎng)物, 并用玻璃刮刀將菌刮下。采用平板菌落計數(shù)法測定指示菌液所含活菌數(shù), 并將其稀釋成密度為1×105CFU/mL的菌液, 備用。

(3)抑菌活性篩選: 采用濾紙片擴散法[6], 測定化合物的抑菌活性。以氯霉素為陽性對照, 每張濾紙片加樣量20 μg。待測化合物以DMSO溶解, 配制為20 g/L溶液, 每張濾紙片加樣量100 μg。將加樣濾紙片貼于帶菌平板表面, 并以加 DMSO溶劑的濾紙片為陰性對照, 將平板置于37℃ 培養(yǎng)箱培養(yǎng)24h后觀察結(jié)果, 測量并記錄抑菌圈直徑。

2 化合物結(jié)構(gòu)鑒定與抑菌活性

化合物1: 無色粉末,1H-NMR (CDCl3)δH: 3.48(1H, m, Ha-3); 3.52 (1H, m, Hb-3); 1.80 (1H, m, Ha-4);2.01 (1H, m, Hb-4); 2.01 (1H, m, Ha-5); 2.60 (1H, m,Hb-5); 4.07 (1H, t,J= 7.7 Hz, H-6); 5.70 (1H, br s,NH-8); 4.28 (1H, dd,J= 10.6, 2.6 Hz, H-9); 2.79 (1H,dd,J= 14.5, 10.6 Hz, Ha-10); 3.50 (1H, m, Hb-10);7.23 (2H, d,J= 7.2 Hz, H-2′/6′); 7.35 (2H, m, H-3′/5′);7.29 (1H, m,H-4′);13C-NMR (CDCl3)δC: 165.1 (C-1,C); 45.5 (C-3, CH2); 22.5 (C-4, CH2); 28.4 (C-5, CH2);59.1 (C-6, CH); 169.4 (C-7, C); 56.2 (C-9, CH); 36.8(C-10, CH2); 136.0 (C-1′, C); 129.3 (C-2′/6′, CH);129.5 (C3′/5′, CH); 127.5 (C4′, CH)。其波譜數(shù)據(jù)與環(huán)-(S-脯氨酸-S-苯丙氨酸)的文獻報道一致[10]?；衔?的比旋光度值為 [α]= –57.1 (c0.28, EtOH), 與文獻報道值 [α]2D0= –64 (c0.05, EtOH)接近, 表明其 C-6與C-9位絕對構(gòu)型均為S[10-11]。該化合物為首次從Aspergillus sydowii中分離得到。

化合物2: 無色粉末,1H-NMR (CDCl3)δH: 3.53(1H, m, Ha-3); 3.59 (1H, m, Hb-3); 1.90 (1H, m, Ha-4);2.02 (1H, m, Hb-4); 2.13 (1H, m, Ha-5); 2.34 (1H, m,Hb-5); 4.11(1H, t,J= 8.2 Hz, H-6); 5.98 (1H, br s,NH-8); 4.01 (1H, dd,J= 9.6, 3.4 Hz, H-9); 1.52 (1H,ddd,J= 14.5, 9.6, 5.0 Hz, Ha-10); 2.06 (1H, m, Hb-10);1.75 (1H, m , H-11); 0.95 (3H, d,J= 6.6 Hz, H-12);1.00 (3H, d,J= 6.6 Hz, H-13);13C-NMR (CDCl3)δC:166.2 (C-1, C); 45.5 (C-3, CH2); 22.7 (C-4, CH2); 28.1(C-5, CH2); 59.0 (C-6, CH); 170.1 (C-7, C); 53.4 (C-9,CH); 38.7 (C-10, CH2); 24.7 (C-11, CH); 21.2 (C-12,CH3); 23.3 (C-13, CH3)。其波譜數(shù)據(jù)與環(huán)-(S-脯氨酸-S-亮氨酸)的文獻報道一致[10]?；衔?的比旋光度值為[α]= –98.0 (c0.51, EtOH), 與文獻報道值 [α= –108 (c0.10, EtOH)接近, 表明其 C-6與 C-9位絕對構(gòu)型均為S[10-11]。該化合物為首次從Aspergillus sydowii中分離得到。

化合物 3: 無色粉末,1H-NMR (DMSO-d6)δH:7.91 (1H, br s, NH-2); 3.97 (1H, br s, H-3); 7.71 (1H,br s, NH-5); 3.85 (1H, br s, H-6); 2.80 (1H, m, Ha-7);2.45 (1H, dd,J= 13.5, 4.5 Hz, Hb-7); 2.53 (1H, m,Ha-8); 1.85 (1H, dd,J= 13.5, 4.6 Hz, Hb-8); 10.89 (1H,br s, NH-1′); 6.96 (1H, d,J= 2.3 Hz, H-2′); 7.48 (1H, d,J= 8.0 Hz, H-4′); 6.98 (1H, m, H-5′); 7.07 (1H, m,H-6′); 7.32 (1H, d,J= 8.1 Hz, H-7′); 7.17 (2H, d,J=7.5 Hz, H-2′/6′); 6.70 (2H, m, H-3′/5′); 7.15 (1H, m,H-4′);13C-NMR (DMSO-d6)δC: 166.7 (C-1, C); 55.6(C-3, CH); 166.1 (C-4, C); 55.2 (C-6, CH); 39.5 (C-7,CH2); 29.6 (C-8, CH2); 124.3 (C-2′, CH); 108.8 (C-3′,C); 118.7 (C-4′, CH); 118.3 (C-5′, CH); 120.8 (C-6′,CH); 111.2 (C-7′, CH); 136.0 (C-8′, C); 127.5 (C-9′, C);136.5 (C-1′, C); 128.0 (C-2′/6′, CH); 129.6 (C-3′/5′,CH); 126.3 (C-4′, CH)。其波譜數(shù)據(jù)與環(huán)-(S-苯丙氨酸-S-色氨酸)的文獻報道值[12]一致。化合物3的比旋光度值為[α= –156.3 (c0.16, MeOH), 與文獻報道值[α= –174.4 (c0.30, MeOH)接近, 表明其 C-6 與 C-9位絕對構(gòu)型均為S[12]。

化合物 4: 無色粉末,1H-NMR (DMSO-d6)δH:1.66 (1H, m, Ha-2); 1.90 (1H, m, Hb-2); 1.08 (1H, m,Ha-3); 1.30 (1H, m, Hb-3); 1.24 (2H, dd,J= 10.7, 4.4 Hz, H-4); 1.50 (3H, s, H-6); 0.98 (3H, s, Me-1); 0.98(3H, s, Me-5); 7.29 (1H, s, H-2′); 7.30 (1H, d,J= 8.0 Hz, H-5′); 7.30 (1H, d,J= 8.0 Hz, H-6′);13C-NMR(DMSO-d6)δC: 75.0 (C-1, C); 42.1 (C-2, CH2); 18.5(C-3, CH2); 44.0 (C-4, CH2); 68.7 (C-5, C); 28.2 (C-6,CH3); 29.2 (Me-1); 29.2 (Me-8); 133.1 (C-1′, C); 116.8(C-2′, CH); 154.5 (C-3′, C); 136.2 (C-4′, C); 119.5(C-5′, CH); 126.3 (C-6′, CH); 168.0 (C-7′, C)。其波譜數(shù)據(jù)與(1S)-1-(4′-間羥基苯甲酸)-1,1,5,5-二甲基己二醇的文獻報道值[13]一致。

化合物 5: 無色粉末,1H-NMR (DMSO-d6)δH:8.01 (1H, s, H-1); 6.33 (1H, s, H-4); 4.29 (2H, s, H-6);9.04 (1H, s, OH-2); 5.66 (1H, s, OH-6);13C-NMR(DMSO-d6)δC: 139.3 (C-1, CH); 145.7 (C-2, C); 174.0(C-3, C); 110.0 (C-4, CH); 168.1 (C-5, C); 59.5 (C-6,CH2)。其波譜數(shù)據(jù)與曲酸文獻報道值[14]一致?；衔?5對金黃色葡萄球菌有較好抑制活性, 加樣量為100 μg/碟時, 抑菌圈直徑為 10 mm, 陽性對照抑菌圈直徑為 22 mm。該結(jié)果與文獻報道的曲酸對金黃色葡萄球菌抑制活性一致[15-16]。

化合物6: 無色粉末,1H-NMR (CDCl3)δH: 1.93(3H, s, H-1); 5.55 (1H, br s, NH-2); 3.61 (2H, t,J= 6.3 Hz, H-3); 2.98 (2H, t,J= 6.3 Hz, H-4); 8.10 (1H, br s,NH-1′); 7.05 (1H, s, H-2′); 7.61 (1H, d,J= 7.9 Hz,H-4′); 7.13 (1H, m, H-5′); 7.21 (1H, m, H-6′); 7.39 (1H, d,J= 7.9 Hz, H-7′);13C-NMR (CDCl3)δC: 23.4 (C-1,CH3); 170.1 (C-2, C); 39.9 (C-3, CH2); 25.3 (C-4, CH2);122.3 (C-2′, CH); 113.2 (C-3′, C); 118.7 (C-4′, CH);119.6 (C-5′, CH); 122.3 (C-6′, CH); 111.3 (C-7′, CH);136.5 (C-8′, C); 127.4 (C-9′, C)。 其 波 譜 數(shù) 據(jù) 與N-[2-(4-吲哚)乙基]乙酰胺文獻報道值[17]一致。

化合物 7: 無色粉末,1H-NMR (DMSO-d6)δH:1.77 (3H, s, H-1); 3.17 (2H, dt,J= 7.8, 6.1Hz, H-3);2.57 (2H, m, H-4); 6.98 (1H, d,J= 8.5 Hz, H-2′); 6.68(1H, d,J= 8.5 Hz, H-3′); 6.66 (1H, d,J= 8.5 Hz, H-5′);6.96 (1H, d,J= 8.5 Hz, H-6′);13C-NMR (DMSO-d6)δC: 22.6 (C-1, CH3); 169.0 (C-2, C); 40.6 (C-3, CH2);34.4 (C-4, CH2); 129.5 (C-1′, C); 129.4 (C-2′, CH);115.1 (C-3′, CH); 155.6 (C-4′, C); 115.1 (C-5′, CH);129.4 (C-6′, CH)。其波譜數(shù)據(jù)與N-[2-(4-對羥基苯酚)乙基]乙酰胺的文獻報道值[18]一致?；衔?對金黃色葡萄球菌有一定抑制活性, 抑菌圈直徑為9 mm。其抑菌活性為首次報道。

化合物 8: 無色粉末,1H-NMR (DMSO-d6)δH:7.70 (1H, s, H-6); 1.77 (3H, s, Me-5); 6.16 (1H, m,H-1′); 2.06 (2H, m, H-2′); 4.23 (1H, m, H-3′); 3.75 (1H,m, H-4′); 3.57 (1H, m, Ha-5′); 3.54 (1H, m, Hb-5′);13C-NMR (DMSO-d6)δC: 150.4 (C-2, C); 163.7 (C-4,C); 109.2 (C-5, C); 136.0 (C-6, CH); 12.1 (Me-5, CH3);87.2 (C-1′, CH); 39.1 (C-2′, CH2); 70.4 (C-3′, CH);83.7 (C-4′, CH); 61.3 (C-5′, CH2)。其波譜數(shù)據(jù)與胸腺嘧啶脫氧核苷的文獻報道值[19]一致。

化合物 9: 無色粉末,1H-NMR (DMSO-d6)δH:5.69 (1H, d,J= 8.1 Hz, H-5); 7.98 (1H, d,J= 8.1 Hz,H-6); 6.27 (1H, m, H-1′); 2.25 (2H, m, H-2′); 4.39 (1H,m, H-3′); 3.92 (1H, m, H-4′); 3.78 (1H, m, Ha-5′); 3.72(1H, m, Hb-5′);13C-NMR (DMSO-d6)δC: 150.9 (C-2,C); 152.5 (C-4, C); 102.7 (C-5, CH); 142.4 (C-6, CH);89.0 (C-1′, CH); 41.4 (C-2′, CH2); 72.3 (C-3′, CH);86.7 (C-4′, CH); 62.9 (C-5′, CH2)。其波譜數(shù)據(jù)與尿嘧啶脫氧核苷的文獻報道值一致[20]。

化合物10: 無色粉末,1H-NMR (DMSO-d6)δH:5.58 (1H, d,J= 8.0 Hz, H-5); 7.83 (1H, d,J= 8.0 Hz,H-6); 5.76 (1H, d,J= 5.3 Hz, H-1′); 3.96 (1H, t,J=4.6 Hz, H-2′); 4.01 (1H, t,J= 4.6 Hz, H-3′); 3.82 (1H,q,J= 3.1 Hz, H-4′); 3.61 (1H, dd,J= 12.0, 3.1 Hz,Ha-5′); 3.54 (1H, dd,J= 12.0, 3.1 Hz, Hb-5′);13C-NMR(DMSO-d6)δC: 151.6 (C-2, C); 164.5 (C-4, C); 101.7(C-5, CH); 140.3 (C-6, CH); 88.0 (C-1′, CH); 69.8(C-2′, CH); 73.5 (C-3′, CH); 84.7 (C-4′, CH); 60.8(C-5′, CH2)。其波譜數(shù)據(jù)與尿嘧啶核苷的文獻報道值[20]一致。

化合物11: 無色粉末;1H-NMR (CDCl3)δH: 3.96(1H, m, H-3); 6.50 (1H, d,J= 8.5 Hz, H-6); 6.24 (1H,d,J= 8.5 Hz, H-7); 0.81 (3H, s, H-18); 0.88 (3H, s,H-19); 0.99 (3H, d,J= 6.6 Hz, H-21); 5.14 (1H, dd,J= 15.3, 8.3 Hz, H-22); 5.22 (1H, dd,J= 15.3, 7.6 Hz,H-23); 0.81 (3H, d,J= 6.9 Hz, H-26); 0.83 (3H, d,J=7.1 Hz, H-27); 0.90 (3H, d,J= 6.9 Hz, H-28);13C-NMR (CDCl3)δC: 34.7 (C-1, CH2); 30.1 (C-2,CH2); 66.4 (C-3, CH); 39.4 (C-4, CH2); 82.1 (C-5, C);135.2 (C-6, CH); 130.7 (C-7, CH); 79.4 (C-8, C); 51.1(C-9, CH); 37.0 (C-10, C); 20.6 (C-11, CH2); 36.9(C-12, CH2); 44.6 (C-13, C); 51.7 (C-14, CH); 23.4(C-15, CH2); 28.6 (C-16, CH2); 56.2 (C-17, CH); 12.9(C-18, CH3); 18.1 (C-19, CH3); 39.7 (C-20, CH); 20.9(C-21, CH3); 135.4 (C-22, CH); 132.3 (C-23, CH); 42.8(C-24, CH); 33.1 (C-25, CH); 19.6 (C-26, CH3); 19.9(C-27, CH3); 17.5 (C-28, CH3)。其波譜數(shù)據(jù)與過氧化麥角甾醇的文獻報道值[21]一致。

化合物12: 無色粉末,1H-NMR (CDCl3)δH: 3.64(1H, m, H-3); 2.47 (1H, m, Ha-4); 2.28 (1H, m, Hb-4);5.57 (1H, dd,J= 5.6, 2.4 Hz, H-6); 5.38 (1H, m, H-7);0.95 (3H, s, H-18); 0.63 (3H, s, H-19); 1.03 (3H, d,J=6.7 Hz, H-21); 5.23 (1H, dd,J= 15.2, 7.1 Hz, H-22);5.17 (1H, dd,J= 15.3, 7.7 Hz, H-23); 0.82 (3H, d,J=7.2 Hz, H-26); 0.84 (3H, d,J= 7.2 Hz, H-27); 0.92 (3H,d,J= 6.9 Hz, H-28);13C-NMR (CDCl3)δC: 38.4 (C-1,CH2); 32.0 (C-2, CH2); 70.5 (C-3, CH); 40.8 (C-4,CH2); 139.8 (C-5, C); 119.6 (C-6, CH); 116.3 (C-7,CH); 141.3 (C-8, C); 46.3 (C-9, CH); 37.1 (C-10, C);21.1 (C-11, CH2); 39.1 (C-12, CH2); 42.8 (C-13, C);54.6 (C-14, CH); 23.0 (C-15, CH2); 28.3 (C-16, CH2);55.8 (C-17, CH); 12.1 (C-18, CH3); 16.3 (C-19, CH3);40.4 (C-20, CH); 21.1 (C-21, CH3); 135.6 (C-22, CH);132.0 (C-23, CH); 42.8 (C-24, CH); 33.1 (C-25, CH);19.9 (C-26, CH3); 19.6 (C-27, CH3); 17.6 (C-28, CH3)。其波譜數(shù)據(jù)與麥角甾醇的文獻報道值[20]一致。

化合物13: 無色粉末;1H-NMR (CDCl3)δH: 3.73(1H, m, Ha-1); 3.88 (1H, m, Hb-1); 3.87 (1H, m, H-2);4.26 (1H, br s, H-3); 5.50 (1H, dd,J= 15.7, 6.3 Hz,H-4); 5.78 (1H, m, H-5); 2.07 (2H, m, H-6); 2.06 (2H,m, H-7); 5.09 (1H, t,J= 6.1 Hz, H-8); 1.94 (2H, t,J=7.6 Hz, H-10); 1.36 (2H, m, H-11); 1.31-1.25 (16H, br s, H12-H19); 0.87 (3H, t,J= 6.9 Hz, H-20); 1.58 (3H,s, H-21); 4.54 (1H, d,J= 6.5 Hz, H-2′); 5.53 (1H, dd,J= 15.2, 6.7 Hz, H-3′); 5.88 (1H, dt,J= 15.2, 6.7 Hz,H-4′); 2.04 (2H, m, H-5′); 1.36 (2H, m, H-6′); 1.31-1.25(18H, br s, H7′-H15′); 0.87 (3H, t,J= 6.9 Hz, H-16′);13C-NMR (CDCl3)δC: 61.9 (C-1, CH2); 54.7 (C-2, CH);74.0 (C-3, CH); 128.7 (C-4, CH); 134.0 (C-5, CH);32.5 (C-6, CH2); 27.6 (C-7, CH2); 123.1 (C-8, CH);136.2 (C-9, C); 39.7 (C-10, CH2); 28.0 (C-11, CH2);29.7-29.2 (C12-C17, CH2); 31.9 (C-18, CH2); 22.7(C-19, CH2); 14.1 (C-20, CH3); 16.0 (C-21, CH3);173.6 (C-1′, C); 73.2 (C-2′, CH); 127.0 (C-3′, CH);135.9 (C-4′, CH); 32.3 (C-5′, CH2); 29.0 (C-6′, CH2);29.7-29.2 (C7′-C13′, CH2); 31.9 (C-14′, CH2); 22.7(C-15′, CH2); 14.1 (C-16′, CH3)。 其 波 譜 數(shù) 據(jù) 與(2S,2′R,3R,3′E,4E,8E)-N-(2′-羥基-3′-十六烯酰基)-9-甲基-4,8-二十碳二烯-1,3-二醇[22]一致。

化合物 14: 無色粉末;1H-NMR (CD3OH)δH:4.14 (1H, dd,J= 9.3, 5.4 Hz, Ha-1); 3.72 (1H, dd,J=10.2, 3.2 Hz, Hb-1); 3.89 (1H, m, H-2); 4.13 (1H, dd,J= 7.1, 5.5 Hz, H-3); 5.48 (1H, dd,J= 16.0, 7.4 Hz,H-4); 5.73 (1H, dt,J= 15.3, 6.6 Hz, H-5); 2.08 (2H, m,H-6); 2.05 (2H, m, H-7); 5.16 (1H, t,J= 5.7 Hz, H-8);1.98 (2H, t,J= 7.3 Hz, H-10); 1.41 (2H, m, H-11);1.36-1.31 (16H, br s, H12-H19); 0.91 (3H, t,J= 7.0 Hz,H-20); 1.60 (3H, s, H-21); 4.43 (1H, d,J= 5.8 Hz,H-2′); 5.48 (1H, dd,J= 16.3, 6.0 Hz, H-3′); 5.85 (1H,dd,J= 15.3, 7.7 Hz, H-4′); 2.03 (2H, m, H-5′); 1.42(2H, m, H-6′); 1.36-1.31 (18H, br s, H7′-H15′); 0.93(3H, t,J= 7.0 Hz, H-16′); 4.28 (1H, d,J= 7.7 Hz,H-1″); 3.20 (1H, t,J= 8.1 Hz, H-2″); 3.36 (1H, t,J=8.8 Hz, H-3″); 3.31 (1H, m, H-4″); 3.30 (1H, m, H-5″);3.89 (1H, br d,J= 11.7 Hz, Ha-6″); 3.71 (1H, dd,J=11.7, 4.3 Hz, Hb-6″);13C-NMR (CD3OH)δC: 69.7 (C-1,CH2); 54.7 (C-2, CH); 72.9 (C-3, CH); 131.0 (C-4, CH);134.5 (C-5, CH); 33.0 (C-6, CH2); 28.8(C-7, CH2);124.9 (C-8, CH); 136.8 (C-9, C); 40.8 (C-10, CH2);29.1 (C-11, CH2); 30.2-30.8 (C12-C17, CH2); 33.1(C-18, CH2); 23.7 (C-19, CH2); 14.4 (C-20, CH3); 16.2(C-21, CH3); 175.5 (C-1′, C); 74.1(C-2′, CH); 129.1(C-3′, CH); 134.8 (C-4′, CH); 33.4 (C-5′, CH2);30.2-30.8 (C6′-C13′, CH2); 33.1 (C-14′, CH2); 23.7(C-15′, CH2); 14.5 (C-16′, CH3); 104.7 (C-1″, CH);75.0 (C-2″, CH); 78.0 (C-3″, CH); 71.6 (C-4″, CH);78.0 (C-5″, CH); 62.7 (C-6″, CH2)。其波譜數(shù)據(jù)與1-O-β-D-葡萄糖基-(2S,2′R,3R,3′E,4E,8E)-N-(2′-羥基-3′-十六烯酰基)-9-甲基-4,8-二十碳二烯-1,3-二醇的文獻報道值[22]一致。

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