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中药药对“都梁丸”化学成分的研究△

2017-01-14韦玮王力梁丽王逾珍海丽娜杨秀伟

中国现代中药 2017年7期
关键词:乙酸乙酯川芎内酯

韦玮,王力,梁丽,王逾珍,海丽娜,杨秀伟*

(1.北京大学 药学院 天然药物学系,天然药物及仿生药物国家重点实验室,北京 100191;2.长治医学院,山西 长治 046000;3.北京振东光明药物研究院有限公司,北京 100120)

中药药对“都梁丸”化学成分的研究△

韦玮1,王力2a,梁丽2a,王逾珍2a,海丽娜3,杨秀伟1*

(1.北京大学 药学院 天然药物学系,天然药物及仿生药物国家重点实验室,北京 100191;2.长治医学院,山西 长治 046000;3.北京振东光明药物研究院有限公司,北京 100120)

目的:研究中药药对“都梁丸”的化学成分。方法:采用硅胶、高效液相等柱色谱方法进行分离纯化,通过质谱、核磁共振等谱学数据鉴定化合物结构。结果:从“都梁丸”环己烷、乙酸乙酯、正丁醇萃取部位中共鉴定了41个化合物,分别为5-羟甲基糠醛(1),香草醛(2),阿魏酸(3),松柏醛(4),花椒毒素(5),佛手酚(6),镰叶芹二醇(7),洋川芎内酯 H(8),洋川芎内酯I(9),洋川芎内酯D(10),4-羟基-3-丁基苯酞(11),洋川芎内酯A(12),Z-藁本内酯(13),3-丁烯基苯酞(14),新蛇床内酯(15),欧当归内酯A(16),β-谷甾醇(17),异茴芹内酯(18),邻苯二甲酸正丁酯(19),邻苯二甲酸异丁酯(20),川芎二内酯R1(21),白术内酰胺(22),香柑内酯(23),补骨脂素(24),异氧化前胡内酯(25),珊瑚菜内酯(26),独活属醇(27),牧草栓翅芹酮(28),异白当归脑(29),欧前胡素(30),异欧前胡素(31),别异欧前胡素(32),水合氧化前胡内酯 (33),白当归素(34),印枳苷(35),(2′S,3′R)-3′-羟基印枳苷(36),仲-O-β-D-吡喃葡萄糖基白当归素(37),叔-O-β-D-吡喃葡萄糖基白当归素(38),(2′R,3′S)-3′-羟基紫花前胡苷(39),花椒毒酚-8-O-β-D-吡喃葡糖苷(40),腺苷(41)。结论:香豆素以及香豆素类成分应来源于复方中的白芷;藁本内酯类化合物应来源于复方中的川芎。

都梁丸;药对;白芷;川芎;化学成分

单味中药是组成方剂的核心,药有个性之特长,方有合群之妙用。单味中药配伍成复方是中医用药的主要形式,亦是中医治法治则在组方用药上的具体应用,显示了传统医药防病治病的特色。因此,中药物质基础的研究,中药材、饮片应突出全成分分析和稳定性物质基础研究,中药复方应突出药效物质基础研究[1]。在系列研究中,我们报道了“黄连解毒汤”[2-4]、“通脉方”[5-9]、“左金方”和“反左金方”[10-13]、“吴茱萸汤(精制吴茱萸胶囊)”[14-16]等物质基础的研究;“都梁丸”是由白芷和川芎以4∶1质量配比组成的蜜丸剂,已被《中华人民共和国药典》2015版收载[17]。方中以白芷为君药,辛温发散、芳香走窜,善入头部祛风散寒、通窍止痛;辅以辛散之川芎活血行气散风,使风除寒散、气血和调、祛风散寒、活血止痛。二味药合用,共奏祛风散寒、活血通络之功。本方治疗风寒之邪上犯头部、清阳之气受阻、气血凝滞引起的头痛。以传统蜜丸为基础的“都梁滴丸”现代中药已开发上市。现代药理学研究证明“都梁丸”具有显著的镇痛作用,可能与影响痛觉调节的神经递质5-羟色胺(5-HT)和5-羟吲哚乙酸(5-HIAA)等的含量有关[18-19];与抑制外周组织c-fos 基因的表达有关[20]。本文报道“都梁丸”物质基础研究。

1 仪器与材料

1.1 仪器

Bruker AV III 400型核磁共振波谱仪(Bruker BioSpin AG Facilities,Fällanden,Switzerland),四甲基硅烷为内标;Finnigan TRACE 2000 GC-MS质谱仪(EI-MS;Thermo Finnigan,San Jose,CA,USA);MDS SCIEX API QSTAR型质谱仪(ESI-TOF-MS;Applied Biosystems/MDS Sciex.,Foster City,CA,USA);LC 3000半制备型高效液相色谱(SP-HPLC)仪系统(北京创新通恒科技有限公司),配置P3050二元泵,CXTH-3000色谱工作站;中压液相色谱柱(46 cm×2.6 cm,北京元宝山色谱科技有限公司);柱色谱硅胶(200~300目,青岛海洋化工厂);Sephadex LH-20为Pharmacia公司产品;MCI-gel CHP 20(75~150 μ)为日本三菱化学产品;GF254薄层色谱(TLC)硅胶板分别为青岛海洋化工厂和Merck公司(Darmstadt,Germany)产品。分析纯乙醇、甲醇、乙酸乙酯、三氯甲烷、丙酮、环己烷等为北京化工厂产品。

1.2 材料

杭白芷药材于2012年8月采自杭白芷道地产地浙江省磐安县深泽乡仰头村,为伞形科植物Angelicadahuricacv.Hangbaizhi的干燥根;川芎药材于2015年5月采自四川省彭州市敖平镇兴全村,为伞形科藁本属植物LigusticumchuanxiongHort.的干燥根茎。杭白芷(HBZ201208)和川芎(20150520CXR)药材凭证标本存放在北京大学天然药物及仿生药物国家重点实验室,均经本文作者之一杨秀伟教授鉴定为《中华人民共和国药典》2015版规定的正品。

2 提取与分离

都梁丸药材粉末(10 kg,其中杭白芷8 kg,川芎2 kg)用70%乙醇水溶液回流提取,料液比为1∶3,共提取8次,合并提取液,过滤后减压回收有机溶剂至无乙醇味,加适量水,然后依次采用石油醚、乙酸乙酯、正丁醇萃取,萃取液分别旋干得到石油醚萃取部位(120 g)、乙酸乙酯萃取部位(181 g)、正丁醇萃取部位(207 g)。

石油醚萃取部位采用硅胶柱进行分离,以石油醚-乙酸乙酯(50∶11∶1)为洗脱体系进行梯度洗脱,经TLC检识,合并相同的流分,得到12个流分(Fr.1~Fr.12)。Fr.10(12 g)采用硅胶柱(环己烷∶乙酸乙酯=9∶1)进行分离得到4个流分(Fr.10-1~Fr.10-4)。Fr.10-4(1 g)经HPLC制备(甲醇∶水=45∶55)分离得到7个流分(Fr.10-4-1~Fr.10-4-7)。其中Fr.10-4-3为水合氧化前胡内酯(33,tR=30.9 min,5 mg),Fr.10-4-4(300 mg)经HPLC制备(乙腈∶水=15∶85)分离得到独活属醇(27,tR=62.1 min,25 mg)。Fr.10-4-6为洋川芎内酯I (9,tR=49.1 min,15 mg),Fr.10-4-7为白当归素(34,tR=55.1 min,200 mg)。Fr.3(2 g)经HPLC制备(乙腈∶水=60∶40)分离得到3-丁烯基苯酞(14,tR=50.5 min,300 mg)。Fr.4 (1.5 g)采用硅胶柱(环己烷∶乙酸乙酯=9∶1)进行分离得到5个流分(Fr.4-1~Fr.4-5)。Fr.4-2(100 mg)经过HPLC(乙腈∶水=60∶40)制备分离得到新蛇床内酯(15,tR=31.4 min,33 mg)。Fr.4-3(195 mg)采用硅胶柱色谱(乙酸乙酯∶三氯甲烷=1∶99)进行分离得到4个流分(Fr.4-3-1~Fr.4-3-4)。其中Fr.4-3-3 (88 mg)采用硅胶柱色谱(环己烷∶乙酸乙酯=9∶1)进行分离得到2个流分(Fr.4-3-3-1~Fr.4-3-3-2)。Fr.4-3-3-2(45 mg)经HPLC(乙腈∶水=55∶45)制备分离得到洋川芎内酯A(12,tR=41.8 min,11 mg)。Fr.7(5 g)经HPLC制备(甲醇∶水=70∶30)分离得到7个流分(Fr.7-1~Fr.7-7)。Fr.7-5(3 g)采用硅胶柱色谱(环己烷∶乙酸乙酯=9∶1)进行分离得到4个流分(Fr.7-5-1~Fr.7-5-4)。Fr.7-5-2(2 g)经HPLC(乙腈∶水=45∶55)制备分离得到异欧前胡素(31,tR=25.1 min,1.0 g)。Fr.7-2(200 mg)经HPLC(甲醇∶水=55∶45)制备分离得到4-羟基-3-丁基苯酞(11,tR=21.6 min,5 mg)。Fr.7-4(22 mg)经HPLC(甲醇∶水=60∶40)制备分离得到补骨脂素(24,tR=34.5 min,4 mg)。Fr.8(1.5 g)经HPLC(乙腈∶水=60∶40)制备分离得到4个流分(Fr.8-1~Fr.8-4)。Fr.8-3(550 mg)经HPLC(乙腈∶水=45∶55)制备分离得到4个流分(Fr.8-3-1~Fr.8-3-4),欧前胡素(30,tR=81.7 min,330 mg),珊瑚菜内酯(26,tR=102.7 min,22 mg)。

乙酸乙酯萃取部位用硅胶柱色谱分离,以石油醚-乙酸乙酯(100∶00∶1)为洗脱体系进行梯度洗脱,经TLC检识,合并相同的流分,得到10个流分(Fr.1~Fr.10)。Fr.1 (1 g) 经硅胶柱色谱(环己烷∶丙酮=10∶1)等度洗脱,分别得到5-羟甲基糠醛(1,5 mg)、香草醛(2,7 mg)和松柏醛 (4,3 mg)。Fr.2(3 g)经硅胶柱色谱(三氯甲烷∶甲醇=100∶1→10∶1)梯度洗脱,TLC检识后合并得到3个流分(Fr.2-1~3),Fr.2-1(1.5 g)流分采用中压制备得到11个流分(Fr.2-1-1~11)。Fr.2-1-6(200 mg)经HPLC制备纯化得到花椒毒素(5,tR=56 min,5 mg)。Fr.2-1-7(300 mg)经硅胶柱色谱,以环己烷-丙酮(30∶1)进行分离得到异茴芹内酯(18,tR=212 min,8 mg)。Fr.2-1-9(700 mg)经硅胶柱色谱(环己烷∶丙酮=49∶1)得到4个流分(Fr.2-1-9-1~4)。Fr.2-1-9-4(100 mg)经HPLC(乙腈∶水=50∶50)制备分离得到牧草栓翅芹酮(28,tR=26 min,7 mg)。Fr.2-1-10(200 mg)经HPLC(甲醇∶水=45∶55)制备分离得到香柑内酯(23,tR=167 min,5 mg)。Fr.2-1-11(450 mg)经硅胶柱色谱(环己烷∶丙酮=30∶1)得到8个流分(Fr.2-1-11-1~8)。Fr.2-1-11-5(55 mg)经制备HPLC(甲醇∶水=60∶40)分离得到异白当归脑(29,tR=51 min,5 mg)。Fr.3 (1.6 g)经硅胶柱色谱(石油醚∶丙酮=30∶1)等度洗脱得到佛手酚(6,3 mg)、镰叶芹二醇(7,1 g)、洋川芎内酯D(10,15 mg)。Fr.4(4 g)经硅胶柱色谱(石油醚∶乙酸乙酯=10∶1)等度洗脱得到Z-藁本内酯(13,400 mg)、3-丁烯基苯酞(14,100 mg)、欧当归内酯A(16,20 mg)、β-谷甾醇(17,1 g)。Fr.5(600 mg)经HPLC(乙腈∶水=70∶30,检测波长:254 nm,流速:8 mL·min-1,进样0.2 mL)制备,得到川芎二内酯 R1(21,tR=3.1 min,5 mg)、白术内酰胺(22,tR=5.0 min,6 mg)、邻苯二甲酸正丁酯(19,tR=43.1 min,23 mg)、邻苯二甲酸异丁酯(20,tR=45.2 min,13 mg)。Fr.6 (1 g)经硅胶柱色谱(环己烷∶乙酸乙酯=10∶1)等度洗脱得到别异欧前胡素(32,11 mg)和水合氧化前胡内酯(33,33 mg)。

正丁醇萃取部位用硅胶柱进行分段,以三氯甲烷-甲醇(20∶1→1∶1)为洗脱体系进行梯度洗脱,经TLC分析后合并相似组份,共得到4个流分(Fr.1-4)。Fr.4(100 g)经制备HPLC以乙腈水为洗脱体系(10∶90→100∶0)分段得到10个流分(Fr.4.5-1~10)。将以上流分Fr.4.5-1~10经过TLC检视,选择流分进行分离。Fr.4.5-1(2 g)经HPLC(乙腈∶水=10∶90,波长:254 nm,流速:8 mL·min-1,进样0.2 mL)制备分段得到3个流分(Fr.4.5-1-1~3),Fr.4.5-1-3(250 mg)经HPLC(乙腈∶水=15∶85,波长:254 nm,流速:5 mL·min-1,进样0.5 mL)制备分离,进一步纯化得到腺苷(41,tR=13.1 min,55 mg)。Fr.4.5-2(13.5 g)经硅胶柱(三氯甲烷∶甲醇=95∶580∶20)进行分段得到4个流分(Fr.4.5-2-1~4)。Fr.4.5-2-3(7 g)经HPLC(乙腈∶水=20∶80,波长:254 nm,流速:15 mL·min-1,进样1 mL)制备分段得到7个流分(Fr.4.5-2-3-1~7)。Fr.4.5-2-4(950 mg)经HPLC(乙腈∶水=20∶80,波长:254 nm,流速:15 mL·min-1,进样1 mL)制备分段得到4个流分(Fr.4.5-2-4-1~4)。其中,Fr.4.5-2-4-3为印枳苷(35,tR=59.3 min,15 mg)。Fr.4.5-3(1.5 g)经HPLC(甲醇∶酸水=25∶75,波长:254 nm,流速:8 mL·min-1,进样0.1 mL)制备分段得到4个流分(Fr.4.5-3-1~4)。其中Fr.4.5-3-3(400 mg)经HPLC(乙腈∶水=12∶88,波长:320 nm,流速:8 mL·min-1,进样0.2 mL)纯化得到洋川芎内酯H (8,tR=79.3 min,8 mg)。Fr.4.5-4(900 mg)经HPLC (甲醇∶水=30∶70) 制备分离得到2个流分(Fr4.5-4-1~2)。Fr.4.5-4-1与Fr4.5-4.2两个流分均经HPLC (乙腈∶水=15∶85,波长:280 nm,流速:8 mL·min-1,进样0.5 mL),制备纯化,得到(2′R,3′S)-3′-羟基紫花前胡苷(39,tR=53.9 min,30 mg)、花椒毒酚-8-O-β-D-吡喃葡萄糖苷(40,tR=55.1 min,11 mg)。Fr.4.5-5与Fr.4.5-6两个流分合并为Fr.4.5-(5.6)(10 g),此流分经硅胶柱色谱(三氯甲烷∶甲醇=95∶580∶20)进行分段得到6个流分[Fr.4.5-(5.6)-1~6],其中,Fr.4.5-(5.6)-5(300 mg)经HPLC(乙腈∶水=30∶70,波长:320 nm,流速:8 mL·min-1,进样0.2 mL)制备分段,得到5个流分[Fr.4.5-(5.6)-5-1~5]。Fr.4.5-(5.6)-6 再经HPLC(乙腈∶水=15∶85,波长:254 nm,流速:15 mL·min-1,进样0.5 mL)制备分段,得到5个流分[Fr.4.5-(5.6)-6-1~5]。Fr.4.5-7(600 mg)经HPLC (甲醇∶水=45∶55,波长:254 nm,流速:10 mL·min-1,进样0.4 mL)制备分段,得到异氧化前胡内酯(25,tR=57.3 min,27 mg)、仲-O-β-D-吡喃葡萄糖基白当归素(37,tR=61.4 min,7 mg)、叔-O-β-D-吡喃葡萄糖基白当归素(38,tR=64.1 min,5 mg)。Fr.4.5-7-1(100 mg)经HPLC(乙腈∶水=17∶83,波长:254 nm,流速:8 mL·min-1,进样0.2 mL)纯化,得到阿魏酸(3,tR=9.3 min,50 mg)和(2′S,3′R)-3′-羟基印枳苷(36,tR=15 min,5 mg)。

3 化合物结构鉴定

化合物1:黄色油状物;EI-MSm/z126 [M]+;1H-NMR (400 MHz,DMSO-d6)δ:9.50 (1H,s,CHO),7.47 (1H,d,J=3.4 Hz,H-3),6.59 (1H,d,J=3.4 Hz,H-4),4.50 (2H,s,CH2OH);13C-NMR (100 MHz,DMSO-d6)δ:178.0 (CHO),162.2 (C-5),151.8 (C-2),124.5 (C-3),109.9 (C-4),56.0 (CH2OH)。NMR数据与文献报道一致[21],故鉴定化合物1为5-羟甲基糠醛(5-hydroxymethylfurfural)。

化合物2:白色固体(甲醇);mp 80 ℃;EI-MSm/z152 [M]+;1H-NMR (CDCl3,400 MHz)δ:9.83 (1H,s,CHO),7.44 (1H,d,J=1.8 Hz,H-2),7.42 (1H,dd,J=8.5,1.8 Hz,H-2),7.05 (1H,d,J=8.5 Hz,H-5),6.16 (1H,s,OH),3.97 (3H,s,OCH3)。NMR数据与文献报道一致[22],故鉴定化合物2为香草醛(vanillin)。

化合物3:无色油状物;EI-MSm/z194 [M]+;1H-NMR (400 MHz,DMSO-d6)δ:7.49 (1H,d,J=15.9 Hz,H-7),7.27 (1H,d,J=1.6 Hz,H-2),7.07 (1H,dd,J=8.1,1.6 Hz,H-6),6.79 (1H,d,J=8.1 Hz,H-5),6.37 (1H,d,J=15.9 Hz,H-8),3.81 (3H,s,3-OCH3);13C-NMR (100 MHz,DMSO-d6)δ:168.1 (C-9),149.1 (C-3),148.0 (C-4),144.4 (C-7),125.9 (C-1),122.8 (C-8),115.9 (C-6),115.6 (C-5),112.2 (C-2),55.7 (3-OCH3)。NMR数据与文献报道一致[23],故鉴定化合物3为阿魏酸(ferulic acid)。

化合物4:黄色固体(乙酸乙酯);mp 372 ℃;EI-MSm/z178 [M]+;1H-NMR (CDCl3,400 MHz)δ:9.65 (1H,d,J=7.7 Hz,CHO),7.40 (1H,d,J=15.8 Hz,H-7),7.13 (1H,dd,J=8.2,1.9 Hz,H-5),7.07 (1H,d,J=1.9 Hz,H-3),6.96 (1H,d,J=8.2 Hz,H-6),6.60 (1H,dd,J=15.8,7.7 Hz,H-8),5.97 (1H,s,OH),3.95 (3H,s,OCH3);13C-NMR (CDCl3,100 MHz)δ:196.4 (C-9),156.1 (C-7),150.9 (C-1),149.8 (C-2),127.2 (C-8),127.0 (C-4),125.1 (C-5),116.8 (C-6),111.5 (C-3),56.1 (OCH3)。NMR数据与文献报道一致[24],故鉴定化合物4为松柏醛(coniferyl aldehyde)。

化合物5:无色针晶(乙酸乙酯);mp 150 ℃;EI-MSm/z216 [M]+;1H-NMR (CDCl3,400 MHz)δ:7.77 (1H,d,J=9.6 Hz,H-4),7.69 (1H,d,J=2.2 Hz,H-2′),7.35 (1H,s,H-5),6.82(1H,d,J=2.2 Hz,H-3′),6.38 (1H,d,J=9.6 Hz,H-3),4.30 (3H,s,8-OCH3);13C-NMR (CDCl3,100 MHz)δ:160.6 (C-2),147.9 (C-7),146.8 (C-2′),144.4 (C-4),143.2 (C-8a),133.0 (C-8),126.3 (C-6),116.7 (C-4a),115.0 (C-3),113.0 (C-5),106.9 (C-3′),61.5 (8-OCH3)。NMR数据与文献报道一致[25],故鉴定化合物5为花椒毒素(xanthotoxin)。

化合物6:无色针晶(乙酸乙酯);mp 276 ℃;EI-MSm/z202 [M]+;1H-NMR (400 MHz,DMSO-d6)δ:11.31 (1H,s,5-OH),8.26 (1H,d,J=9.8 Hz,H-4),7.91 (1H,d,J=2.3 Hz,H-2′),7.20 (1H,d,J=2.3 Hz,H-3′),7.16 (1H,s,H-8),6.27 (1H,d,J=9.8 Hz,H-3)。NMR数据与文献报道一致[26],故鉴定化合物6为佛手酚(bergaptol)。

化合物7:淡黄色油状物;EI-MSm/z260 [M]+;1H-NMR (CDCl3,400 MHz)δ:5.89 (1H,ddd,J=17.0,10.1,5.4 Hz,H-2),5.58~5.54 (1H,m,H-10),5.48 (1H,d,J=8.2 Hz,H-9),5.41 (1H,d,J=17.1 Hz,Hb-1),5.20 (1H,d,J=10.1 Hz,Ha-1),5.15 (1H,d,J=8.2 Hz,H-8),4.89 (1H,d,J=5.4 Hz,H-3),2.05 (2H,q,J=7.4 Hz,H-11),1.30~1.39 (m,2H),1.20~1.30 (m,8H),0.84 (3H,t,J=6.8 Hz,H-17);13C-NMR (CDCl3,100 MHz)δ:135.9 (C-2),134.2 (C-10),127.8 (C-9),117.2 (C-1),79.8 (C-4),78.4 (C-7),70.2 (C-5),68.7 (C-6),63.2 (C-3),58.4 (C-8),31.8 (C-15),29.3 (C-12),29.2 (C-13),29.1 (C-14),27.7 (C-11),22.6 (C-16),14.1 (C-17)。NMR数据与文献报道一致[27],故鉴定化合物7为镰叶芹二醇(falcarindiol)。

化合物8:无色油状物;EI-MSm/z224[M]+;1H-NMR (CDCl3,400 MHz)δ:5.27 (1H,t,J=7.7 Hz,H-8),4.57 (1H,brs,H-7),3.95 (1H,brs,H-6),2.61 (1H,m,Hb-4),2.40 (1H,m,Ha-4),2.33 (2H,m,H-9),2.04 (1H,brs,Hb-5),1.83 (1H,brs,Ha-5),1.46 (2H,sext,H-10),0.92 (3H,t,J=7.2 Hz,H-11);13C-NMR (CDCl3,100 MHz)δ:169.5 (C-1),153.8(C-3),148.3 (C-3a),125.9 (C-7a),114.5 (C-8),68.2 (C-6),62.8 (C-7),28.2 (C-9),25.3 (C-5),22.3 (C-10),19.1 (C-4),13.9 (C-11)。NMR数据与文献报道一致[28],故鉴定化合物8为洋川芎内酯 H (senkyunolide H)。

化合物9:淡黄色油状物;EI-MSm/z224[M]+;1H-NMR (CDCl3,400 MHz)δ:0.94 (3H,t,J=7.4 Hz,H-11),1.51 (2H,m,H-10),1.91 (1H,m,H-5),2.10 (H,m,H-5),2.31 (2H,q,H-9),2.39 (2H,m,H-4),3.98 (1H,m,H-6),4.48 (1H,d,J=5.2 Hz,H-7),5.27 (1H,t,J=8.4 Hz,H-8);13C-NMR (CDCl3,100 MHz)δ:169.3 (C-1),151.9 (C-3),151.1 (C-3a),125.8 (C-7a),115.5 (C-8),71.1 (C-6),67.6 (C-7),28.0 (C-9),26.8 (C-5),22.1 (C-10),19.6 (C-4),13.8 (C-11)。NMR数据与文献报道一致[28],故鉴定化合物9为洋川芎内酯I (senkyunolide I)。

化合物10:淡黄色油状物;EI-MSm/z222[M]+;1H-NMR (CDCl3,400 MHz)δ:6.26 (1H,d,J=9.4 Hz,H-7),6.09 (1H,m,H-6),5.34 (1H,s,OH),2.67 (2H,m,H-9),2.27 (2H,m,H-6),2.48 (1H,m,H-5),2.23 (1H,m,H-5),1.65 (2H,m,H-10),0.92 (3H,t,J=7.3 Hz,H-11);13C-NMR (CDCl3,100 MHz)δ:202.7 (C-9),168.7 (C-1),155.1 (3a),131.8 (C-6),129.07,116.3 (C-7),102.8 (C-3),36.6 (C-9),22.6 (C-6),19.1 (C-4),17.1 (C-10),13.6 (C-11)。NMR数据与文献报道一致[29],故鉴定化合物10为洋川芎内酯D (senkyunolide D)。

化合物11:白色结晶(甲醇);mp 190 ℃;ESI-MSm/z207.1 [M+H]+;1H-NMR (CDCl3,400 MHz)δ:7.48 (1H,d,J=7.6 Hz,H-5),7.37 (1H,t,J=7.6 Hz,H-6),7.01 (1H,d,J=7.6 Hz,H-7),5.68 (1H,s,7-OH),5.56 (1H,dd,J=7.9,2.9 Hz,H-3),2.30 (1H,m,H-8),1.78 (1H,m,H-8),1.48~1.28 (4H,m,H-9,10),0.90 (3H,t,J=6.5 Hz,H-11);13C-NMR(CDCl3,100 MHz)δ:171.4 (C-1),136.4 (C-3a),152.5 (C-4),120.2 (C-5),130.4 (C-6),115.9 (C-7),127.9 (C-7a),80.9 (C-3),32.7 (C-8),27.0 (C-9),22.9 (C-10),14.0 (C-11)。NMR数据与文献报道一致[30],故鉴定化合物11为4-羟基-3-丁基苯酞(4-hydroxy-3-butyl-phthalide)。

化合物12:无色油状物;EI-MSm/z192 [M]+;1H-NMR (CDCl3,400 MHz)δ:6.20 (1H,d,J=9.7 Hz,H-7),5.95~5.86 (1H,m,H-6),4.92 (1H,dd,J=7.3,3.5 Hz,H-3),2.55~2.39 (2H,m,H-4),2.55~2.39 (2H,m,H-5),1.87 (1H,m,H-8),1.59~1.46 (1H,m,H-8),1.47~1.31 (2H,m,H-9),1.47~1.31 (2H,m,H-10),0.90 (3H,t,J=7.0 Hz,H-11);13C-NMR (CDCl3,100 MHz)δ:171.4 (C-1),161.5 (C-3a),128.4 (C-6),124.7 (C-7a),117.0 (C-7),82.6 (C-3),32.1 (C-8),26.9 (C-9),22.6 (C-5),22.4 (C-10),20.9 (C-4),14.0 (C-11)。NMR数据与文献报道一致[31],故鉴定化合物12为洋川芎内酯A(senkyunolide A)。

化合物13:淡黄色油状物;EI-MSm/z190 [M]+;1H-NMR (CDCl3,400 MHz)δ:6.25 (1H,dt,J=9.7 1.5 Hz,H-7),5.95 (1H,m,H-6),5.18 (1H,t,J=8.0 Hz,H-8),2.57 (2H,t,J=13.5 Hz,H-5),2.55~2.39 (2H,m,H-4),2.33 (2H,m,H-9),1.47~1.31 (2H,m,H-10),0.92 (3H,t,J=7.5 Hz,H-11);13C-NMR (CDCl3,100 MHz)δ:167.4 (C-1),148.5 (C-3a),147.4 (C-3),129.7 (C-6),123.7 (C-7a),117.0 (C-7),112.1 (C-8),28.2 (C-9),22.4 (C-10),22.1 (C-5),18.9 (C-4),14.0 (C-11)。NMR数据与文献报道一致[32],故鉴定化合物13为Z-藁本内酯(Z-ligustilide)。

化合物14:淡黄色油状物;ESI-MSm/z189 [M+H]+;1H-NMR (CDCl3,400 MHz)δ:6.13 (1H,d,J=8.1 Hz,H-7),6.00 (1H,m,H-6),5.10 (1H,t,J=7.6 Hz,H-8),2.60 (2H,m,H-9),2.54 (2H,m,H-5),2.39 (2H,m,H-4),1.40 (2H,m,H-10),0.96 (3H,t,J=7.2 Hz,H-11)。NMR数据与文献报道一致[33],故鉴定化合物14为3-丁烯基苯酞(3-butylidenephthalide)。

化合物15:无色油状物;EI-MSm/z194 [M]+;1H-NMR (CDCl3,400 MHz)δ:6.74 (1H,dt,J=3.2,3.2 Hz,H-7),3.94 (1H,ddd,J=8.8,7.2,5.6 Hz,H-3),2.53~2.41 (1H,m,H-3a),2.32 (1H,m,H-6),2.24~2.11 (1H,m,H-6),2.08~2.00 (1H,m,H-4),1.91 (1H,dddd,J=13.9,6.8,3.3,3.1 Hz,H-5),1.82~1.65 (2H,m,H-8),1.60~1.44 (2H,m,H-5,9),1.44~1.26 (2H,m,H-9,10),1.24~1.08 (1H,m,H-4),0.89 (3H,t,J=7.1 Hz,H-11);13C-NMR (CDCl3,100 MHz)δ:170.2 (C-1),135.2 (C-7),131.2 (C-7a),85.4 (C-3),43.1 (C-3a),34.4 (C-8),27.6 (C-9),25.4 (C-4),25.0 (C-6),22.6 (C-10),20.8 (C-5),13.9 (C-11)。NMR数据与文献报道一致[34],故鉴定化合物15为新蛇床内酯(neocnidilide)。

化合物16:白色固体(丙酮);mp 114 ℃;ESI-MSm/z381.2 [M+H]+;1H-NMR (CDCl3,400 MHz)δ:7.33 (1H,d,J=6.6 Hz,H-7′),5.05 (1H,t,J=7.9 Hz,H-8),4.97 (1H,t,J=7.4 Hz,H-8′),3.23 (1H,d,J=8.9 Hz,H-7),2.97 (1H,m,H-6′),2.53 (1H,t,J=7.9 Hz,H-6),2.26 (2H,m,H-9),2.19~2.11 (2H,m,H-9′),0.91 (3H,t,J=7.3,H-11),0.90 (3H,t,J=7.3,H-11′);13C-NMR (CDCl3,100 MHz)δ:168.5 (C-1),165.0 (C-1′),155.1 (C-3a),150.6 (C-3′),148.1 (C-3),142.2 (C-7′),134.2 (C-7′a),126.6 (C-7a),112.1 (C-8),108.6 (C-8′),47.7 (C-3′a),41.6 (C-7),41.6 (C-6′),38.4 (C-6),31.1 (C-4′),29.8 (C-3),28.1 (C-9),27.5 (C-9′),25.8 (C-5′),22.5 (C-10),22.4 (C-10′),19.8 (C-4),14.0 (C-11),13.9 (C-11′)。NMR数据与文献报道一致[33],故鉴定化合物16为欧当归内酯A (levistolide A)。

化合物17:白色粉末(甲醇),与对照品β-谷甾醇TLC分析,3种展开剂展开,色谱行为一致,故鉴定为β-谷甾醇(β-sitosterol)。

化合物18:黄色粉末(乙酸乙酯);mp 152 ℃;EI-MSm/z246 [M]+;1H-NMR (CDCl3,400 MHz)δ:8.12 (1H,d,J=9.8 Hz,H-4),7.62 (1H,d,J=2.2 Hz,H-2′),6.99 (1H,d,J=2.2 Hz,H-3′),6.29 (1H,d,J=9.8 Hz,H-3),4.17 (3H,s,OCH3),4.17 (3H,s,OCH3);13C-NMR (CDCl3,100 MHz)δ:60.9 (5-OCH3),61.7 (8-OCH3),105.1 (C-3′),107.7 (C-10),113.0 (C-3),114.9 (C-6),128.3 (C-8),139.4 (C-4),143.7 (C-9),144.3 (C-5),145.1 (C-2′),150.0 (C-7),160.4 (C-2)。NMR波谱数据与文献报道一致[25],故鉴定化合物18为异茴芹内酯(isopimpinellin)。

化合物19:无色油状物;EI-MSm/z278 [M]+;1H-NMR (400 MHz,CDCl3)δ:7.72 (2H,dd,J=5.3,3.5 Hz,H-3,6),7.53 (2H,dd,5.3,3.5 Hz,H-4,5),4.30 (4H,t,J=6.7 Hz,H-9,9′),1.72 (4H,m,H-11,11′),1.44 (4H,m,H-10,10′),0.96 (6H,t,J=7.4 Hz,12,12′-CH3,)。NMR数据与文献报道一致[35],故鉴定化合物19为邻苯二甲酸正丁酯(dibutyl-O-phthalate)。

化合物20:无色油状物;EI-MSm/z278 [M]+;1H-NMR (400 MHz,CDCl3)δ:7.72 (2H,dd,J=5.3,3.5 Hz,H-3,6),7.53 (2H,dd,J=5.5,3.5 Hz,H-4,5),4.09 (4H,d,J=6.7 Hz,H-1′,1″),2.04 (2H,m,H-2′,2″),0.99 (12H,d,J=6.7 Hz,H-3′,3″,4′,4″)。NMR数据与文献报道一致[35],故鉴定化合物20为邻苯二甲酸异丁酯(diisobutyl phthalate)。

化合物21:淡黄色油状物;ESI-MSm/z379.1 [M+H]+;1H-NMR (400 MHz,CDCl3)δ:7.76 (1H,d,J=7.6 Hz,H-7),7.57 (1H,d,J=7.0 Hz,H-7′),7.53 (1H,t,J=7.7 Hz,H-5),7.41 (1H,t,J=7.7 Hz,H-6),7.15 (1H,d,J=7.7 Hz,H-4),3.70 (3H,s,OCH3),2.90 (1H,m,H-6′),2.62 (1H,m,H-4′),2.44 (2H,m,H-8′),2.13 (1H,m,H-5′),1.83 (1H,m,H-8),1.75 (1H,m,H-4′),1.53 (1H,m,H-5′),1.50 (2H,m,H-9),1.23 (2H,m,H-9′),1.13 (2H,m,H-10′),0.83 (3H,J=7.2 Hz,H-11′),0.76 (3H,t,J=7.2 Hz,H-11);13C-NMR (100 MHz,CDCl3)δ:207.1 (C-3′),170.4 (C-1),165.4 (C-1′),155.0 (C-3a),148.6 (C-7′),136.2 (C-7a′),133.9 (C-5),128.5 (C-6),125.7 (C-7a),124.6 (C-7),122.0 (C-4),89.0 (C-3),58.2 (C-3a′),52.1 (OCH3),49.1 (C-8),40.9 (C-8′),33.8 (C-6′),28.1 (C-9),26.3 (C-9),25.6 (C-9′),22.1 (C-10′),20.7 (C-10),17.9 (C-5′),14.0 (C-11),13.8 (C-11′)。NMR数据与文献报道一致[36],故鉴定化合物21为川芎二内酯R1(chuanxiongdiolide R1)。

化合物22:白色粉末(甲醇);mp 120~123 ℃;ESI-MSm/z230.2 [M+H]+。1H-NMR (400 MHz,CDCl3),δ:7.50 (1H,s,N-H),4.63 (1H,brs,Ha-15),4.89 (1H,brs,Hb-15),5.45 (1H,s,H-9),2.63 (1H,dd,J=3.8,16.4 Hz,Hb-6),2.46 (1H,d,J=13.4 Hz,Ha-6),2.36 (1H,m,H-3),2.34 (1H,m,H-1),2.04 (1H,m,H-1),1.64 (2H,m,H-2),1.61 (1H,m,H-3),1.58 (1H,m,H-5),1.87 (3H,s,H-13),0.91 (3H,s,H-14);13C-NMR (CDCl3,100 MHz)δ:173.3 (C-12),148.8 (C-4),141.8 (C-8),135.4 (C-7),124.9 (C-11),120.9 (C-9),107.2 (C-15),49.2 (C-5),39.5 (C-1),38.2 (C-10),36.4 (C-3),23.3 (C-6),22.5 (C-2),18.7 (C-14),8.3 (C-13)。NMR 数据与文献报道一致[37],故鉴定化合物22为白术内酰胺(atractylenolactam)。

化合物23:淡黄色结晶(甲醇);mp 188 ℃;EI-MSm/z216 [M]+;1H-NMR (CDCl3,400 MHz)δ:8.13 (1H,d,J=9.8 Hz,H-4),7.59 (1H,d,J=2.2 Hz,H-2′),7.10 (1H,s,H-8),7.02 (1H,d,J=2.2 Hz,H-3′),6.25 (1H,d,J=9.8 Hz,H-3),4.27 (3H,s,5-OCH3);13C-NMR (CDCl3,100 MHz)δ:161.2 (C-2),158.3 (C-7),152.7 (C-8a),149.5 (C-5),144.7 (C-2′),139.2 (C-4),112.7 (C-6),112.5 (C-3),106.4 (C-4a),105.0 (C-3′),93.8 (C-8),60.1 (5-OCH3)。NMR数据与文献报道一致[38],故鉴定化合物23为香柑内酯(bergapten)。

化合物24:黄色固体;mp 186 ℃;EI-MSm/z186 [M]+;1H-NMR (CDCl3,400 MHz)δ:7.80 (1H,d,J=9.6 Hz,H-4),7.69 (1H,d,J=2.2 Hz,H-2′),7.68 (1H,s,H-5),7.47 (1H,s,H-8),6.83 (1H,d,J=2.2 Hz,H-3′),6.38 (1H,d,J=9.6 Hz,H-3)。NMR数据与文献报道一致[25],故鉴定化合物24为补骨脂素(psoralen)。

化合物25:白色粉末(丙酮);mp 146 ℃;EI-MSm/z286 [M]+;1H-NMR (CDCl3,400 MHz)δ:8.32 (1H,d,J=9.8 Hz,H-4),7.61 (1H,d,J=2.4 Hz,H-2′),7.17 (1H,s,H-8),6.84 (1H,d,J=2.4 Hz,H-3′),6.32 (1H,d,J=9.8 Hz,H-3),5.10 (2H,s,H-1″),2.86 (1H,q,J=8.0 Hz,H-3″),1.18 (6H,d,J=6.9 Hz,3″-CH3);13C-NMR (CDCl3,100 MHz)δ:208.9 (C-2″),161.1 (C-2),158.1 (C-7),152.7 (C-8a),148.0 (C-5),145.6 (C-2′),139.4 (C-4),113.7 (C-6),113.4 (C-3),107.6 (C-4a),104.2 (C-3′),95.2 (C-8),75.1 (C-1″),37.5 (C-3″),18.1 (3″-CH3)。NMR数据与文献报道一致[25],故鉴定化合物25为异氧化前胡内酯(isooxypeucedanin)。

化合物26:淡黄色固体(乙酸乙酯);mp 102 ℃;EI-MSm/z300 [M]+;1H-NMR (CDCl3,400 MHz)δ:8.11 (1H,d,J=9.8 Hz,H-4),7.60 (1H,d,J=2.3 Hz,H-2′),6.99 (1H,J=2.3 Hz,H-3′),6.26 (1H,d,J=9.8 Hz,H-3),5.59 (1H,t,J=7.2 Hz,H-2″),4.83 (2H,d,J=7.2 Hz,H-1″),4.16 (3H,s,5-OCH3),1.72 (3H,s,3″-CH3),1.68 (3H,s,3″-CH3);13C-NMR (CDCl3,100 MHz)δ:160.7 (C-2),150.9 (C-7),145.2 (C-2′),144.5 (C-8a),144.4 (C-5),139.8 (C-3″),139.6 (C-4),126.9 (C-8),119.9 (C-2″),114.6 (C-6),112.8 (C-3),107.6 (C-4a),105.2 (C-3′),70.5 (C-1″),60.8 (5-OCH3),25.9 (3″-CH3),18.2 (3″-CH3)。NMR数据与文献报道一致[25],故鉴定化合物26为珊瑚菜内酯(phellopterin)。

化合物27:白色粉末(乙酸乙酯);mp 130 ℃;EI-MSm/z304 [M]+;1H-NMR (CDCl3,400 MHz)δ:7.74 (1H,d,J=9.6 Hz,H-4),7.69 (1H,d,J=2.2 Hz,H-2′),7.35 (1H,s,H-5),6.81 (1H,d,J=2.2 Hz,H-3′),6.34 (1H,d,J=9.6 Hz,H-3),4.73 (1H,dd,J=10.2,2.7 Hz,Hb-1″),4.40 (1H,dd,J=10.2,7.9 Hz,Ha-1″),3.87 (1H,dd,J=7.9,2.7 Hz,H-2″),1.32 (3H,s,3″-CH3),1.28 (3H,s,3″-CH3);13C-NMR (CDCl3,100 MHz)δ:160.4 (C-2),148.0 (C-7),146.9 (C-2′),144.5 (C-4),143.3 (C-8a),131.7 (C-8),126.2 (C-6),116.5 (C-4a),114.8 (C-3),113.8 (C-5),106.9 (C-3′),76.1 (C-2″),75.7 (C-1″),71.7 (C-3″),25.2 (3″-CH3),26.7 (3″-CH3)。NMR数据与文献报道一致[38],故鉴定化合物27为独活属醇(heraclenol)。

化合物28:无色针晶 (石油醚-乙酸乙酯);mp 133 ℃;ESI-MSm/z287.1[M+H]+;1H-NMR (CDCl3,400 MHz)δ:7.76 (1H,d,J=9.6 Hz,H-4),7.66 (1H,d,J=2.0 Hz,H-2′),7.36 (1H,s,H-5),6.81 (1H,d,J=2.0 Hz,H-3′),6.37 (1H,d,J=9.6 Hz,H-3),5.19 (2H,s,H-1″),3.05 (1H,m,H-3″),1.21 (3H,s,3″-CH3),1.20 (3H,s,3″-CH3)。NMR数据与文献报道一致[39],故鉴定化合物28为牧草栓翅芹酮(pabularinone)。

化合物29:白色粉末(甲醇);mp 106 ℃;ESI-MSm/z317.1[M+H]+;1H-NMR (CDCl3,400 MHz)δ:8.08 (1H,d,J=9.8 Hz,H-4),7.59 (1H,d,J=2.3 Hz,H-2′),6.98 (1H,d,J=2.3 Hz,H-3′),6.23 (1H,d,J=9.8 Hz,H-3),5.00 (2H,s,H-1″),4.15 (3H,s,5-OCH3),3.07 (1H,q,J=6.9 Hz,H-3″),1.18 (6H,d,J=6.9 Hz,3″-CH3);13C-NMR (CDCl3,100 MHz)δ:210.3 (C-2″),160.2 (C-2),149.5 (C-7),145.3 (C-2′),144.7 (C-5),143.3 (C-8a),139.5 (C-4),126.4 (C-8),114.7 (C-6),112.8 (C-3),107.5 (C-4a),105.3 (C-3′),75.9 (C-1″),60.8 (5-OCH3),37.0 (C-3″),18.0 (3″-CH3)。NMR数据与文献报道一致[25],故鉴定化合物29为异白当归脑(anhydrobyakangelicin)。

化合物30:无色针晶(乙酸乙酯);mp 102 ℃;ESI-MSm/z271.1[M+H]+;1H-NMR (400 MHz,CDCl3)δ:7.76 (1H,d,J=9.6 Hz,H-4),7.68 (1H,d,J=2.2 Hz,H-2′),7.35 (1H,s,H-5),6.80 (1H,d,J=2.2 Hz,H-3′),6.35 (1H,d,J=9.6 Hz,H-3),5.60 (1H,t,J=7.2 Hz,H-2″),4.99 (2H,d,J=7.2 Hz,H-1″),1.73 (3H,s,3″-CH3),1.71 (3H,s,3″-CH3);13C-NMR (100 MHz,CDCl3)δ:160.7 (C-2),148.7 (C-7),146.8 (C-2′),144.5 (C-4),143.9 (C-8a),139.9 (C-3″),131.8 (C-8),126.0 (C-6),119.9 (C-2″),116.6 (C-4a),114.8 (C-3),113.3 (C-5),106.8 (C-3′),70.3 (C-1″),25.9 (3″-CH3),18.2 (3″-CH3)。NMR数据与文献报道一致[38],故鉴定化合物30为欧前胡素(imperatorin)。

化合物31:无色针晶(乙酸乙酯);mp 108 ℃;ESI-MSm/z271.1[M+H]+;1H-NMR (400 MHz,CDCl3)δ:8.16 (1H,d,J=9.8 Hz,H-4),7.59 (1H,d,J=2.4 Hz,H-2′),7.15 (1H,s,H-8),6.95 (1H,d,J=2.4 Hz,H-3′),6.27 (1H,d,J=9.8 Hz,H-3),5.54 (1H,t,J=7.0 Hz,H-2″),4.92 (2H,d,J=7.0 Hz,H-1″),1.80 (3H,s,3″-CH3),1.70 (3H,s,3″-CH3);13C-NMR (100 MHz,CDCl3)δ:161.4 (C-2),158.3 (C-7),152.8 (C-9),149.1 (C-5),145.0 (C-2′),140.0 (C-3″),139.7 (C-4),119.3 (C-2″),114.4 (C-6),112.7 (C-3),107.7 (C-4a),105.2 (C-3′),94.4 (C-8),69.9 (C-1″),25.9 (3″-CH3),18.4 (3″-CH3)。NMR数据与文献报道一致[38],故鉴定化合物31为异欧前胡素(isoimperatorin)。

化合物32:无色针晶 (甲醇);mp 178 ℃;ESI-MSm/z271.5 [M+H]+;1H-NMR (DMSO-d6,400 MHz)δ:8.18 (1H,d,J=9.8 Hz,H-4),8.04 (1H,br s,H-2′),7.08 (1H,br s,H-3′),6.40 (1H,d,J=9.8 Hz,H-3),5.12 (1H,t,J=6.6 Hz,H-2″),3.72 (2H,d,J=6.6 Hz,H-1″),3.19 (1H,br s,5-OH),1.80 (3H,s,3″-CH3),1.63 (3H,s,3″-CH3);13C-NMR (DMSO-d6,100 MHz)δ:160.2 (C-2),146.9 (C-2′),145.4 (C-7),142.0 (C-5,9),140.9 (C-4),131.2 (C-3″),128.1 (C-6),124.9 (C-8),123.2 (C-2″),113.9 (C-10),113.6 (C-3),106.6 (C-3′),27.4 (3″-CH3),25.1 (C-1″),18.0 (3″-CH3)。NMR数据与文献报道一致[40],故鉴定化合物32为别异欧前胡素(alloisoimperatorin)。

化合物33:淡黄色固体(丙酮);mp 138 ℃;EI-MSm/z304 [M]+;1H-NMR (400 MHz,CDCl3)δ:8.16 (1H,d,J=9.8 Hz,H-4),7.59 (1H,d,J=1.8 Hz,H-2′),7.11 (1H,s,H-8),6.98 (1H,d,J=1.8 Hz,H-3′),6.24 (1H,d,J=9.8 Hz,H-3),4.54 (1H,dd,J=9.7,2.5 Hz,Hb-1″),4.43 (1H,t,J=9.7 Hz,Ha-1″),3.91 (1H,d,J=7.2 Hz,H-2″),1.36 (3H,s,3″-CH3),1.31 (3H,s,3″-CH3);13C-NMR (100 MHz,CDCl3)δ:161.3 (C-2),158.2 (C-7),152.6 (C-8a),148.7 (C-5),145.4 (C-2′),139.3 (C-4),114.3 (C-6),113.1 (C-3),107.4 (C-4a),104.9 (C-3′),94.8 (C-8),76.7 (C-3″),74.6 (C-2″),71.8 (C-1″),26.8 (3″-CH3),25.3 (3″-CH3)。NMR数据与文献报道一致[38],故鉴定化合物33为水合氧化前胡内酯 (oxypeucedanin hydrate)。

化合物34:白色粉末(甲醇);mp 118 ℃;ESI-MSm/z357.1 [M+Na]+;1H-NMR (400 MHz,CDCl3)δ:8.12 (1H,d,J=9.8 Hz,H-4),7.63 (1H,d,J=2.3 Hz,H-2′),7.02 (1H,d,J=2.3 Hz,H-3′),6.29 (1H,d,J=9.8 Hz,H-3),4.60 (1H,dd,J=10.2,2.6 Hz,Hb-1″),4.26 (1H,dd,J=10.2,7.9 Hz,Ha-1″),4.19 (3H,s,5-OCH3),3.83 (1H,dd,J=7.9,2.6 Hz,H-2″),1.32 (3H,s,3″-CH3),1.28 (3H,s,3″-CH3);13C-NMR (100 MHz,CDCl3)δ:160.3 (C-2),150.3 (C-7),145.4 (C-2′),145.0 (C-5),144.1 (C-8a),139.6 (C-4),126.9 (C-8),114.7 (C-6),113.0 (C-3),107.7 (C-4a),105.5 (C-3′),76.2 (C-2″),76.1 (C-1″),71.7 (C-3″),60.9 (5-OCH3),26.8 (3″-CH3),25.2 (3″-CH3)。NMR数据与文献报道一致[25],故鉴定化合物34为白当归素(byakangelicin)。

化合物35:白色粉末(乙酸乙酯);mp 260 ℃;ESI-MSm/z431 [M+Na]+,407 [M-H]-;1H-NMR (DMSO-d6,400 MHz)δ:7.95 (1H,d,J=9.5 Hz,H-4),7.48 (1H,s,H-5),6.82 (1H,s,H-8),6.22 (1H,d,J=9.5 Hz,H-3),4.82~4.89 (4H,m,H-2′,4″-OH,3″-OH,2″-OH),4.40 (1H,d,J=7.8 Hz,H-1″),4.30 (1H,t,J=5.7 Hz,6″-OH),3.44~3.33 (2H,m,Hb-6″,Ha-6″),3.33~3.03 (5H,m,H-3′,3″,4″,5″),2.88~2.83 (1H,m,H-2″),1.26 (3H,s,4′-CH3),1.23 (3H,s,4′-CH3);13C-NMR (DMSO-d6,100 MHz)δ:163.2 (C-7),160.6 (C-2),155.1 (C-9),144.8 (C-4),125.7 (C-6),124.0 (C-5),112.3 (C-10),111.4 (C-3),97.4 (C-1″),96.9 (C-8),90.2 (C-2′),77.0 (C-4′),77.0 (C-5″),76.6 (C-3″),73.6 (C-2″),70.1 (C-4″),61.0 (C-6″),28.9 (C-3′),23.2 (4′-CH3),21.9 (4′-CH3)。NMR数据与文献报道一致[40],故鉴定化合物35为印枳苷[(-)-marmesinin]。

化合物36:白色针状结晶(甲醇);mp 258 ℃;ESI-MSm/z447 [M+Na]+;1H-NMR (DMSO-d6,400 MHz)δ:8.00 (1H,d,J=9.6 Hz,H-4),7.66(1H,s,H-5),6.96 (1H,s,H-8),6.28 (1H,d,J=9.6 Hz,H-3),5.20~5.27 (2H,m,H-3′,3′-OH),4.99 (1H,d,J=3.1 Hz,4″-OH),4.89 (1H,br s,3″-OH),4.86 (1H,br s,2″-OH),4.52~4.55 (2H,m,H-1″,H-2′),4.31 (1H,br s,6″-OH),3.36 (1H,overlap,Ha-6″,Hb-6″),3.15 (1H,m,H-5″),3.06 (2H,br s,H-3″,4″),2.88 (1H,d,J=7.2 Hz,H-2″),1.47 (6H,s,2×4′-CH3);13C-NMR (DMSO-d6,100 MHz)δ:162.4 (C-7),160.4 (C-2),156.1 (C-9),144.9 (C-4),128.6 (C-6),125.7 (C-5),112.9 (C-10),111.8 (C-3),97.7 (C-1″),97.3 (C-8),91.9 (C-2′),77.5 (C-3′),76.9 (C-3″),76.7 (C-5″),73.5 (C-2″),70.1 (C-4″),69.8 (C-4′),60.8 (C-6″),24.6 (4′-CH3),22.8 (4′-CH3)。NMR数据与文献报道一致[41],故鉴定化合物36为(2′S,3′R)-3′-羟基印枳苷[(2′S,3′R)-3′-hydroxymarmesinin]。

化合物37:白色固体(乙腈-水);mp 925 ℃;ESI-MSm/z496 [M+H]+;1H-NMR (DMSO-d6,400 MHz)δ:8.18 (1H,d,J=9.8 Hz,H-4),8.10 (1H,d,J=2.3 Hz,H-2′),7.34 (1H,d,J=2.3 Hz,H-3′),6.34 (1H,d,J=9.8 Hz,H-3),5.25-4.93 (3H,br s,2‴-OH,3‴-OH,4‴-OH),4.50-4.43 (4H,m,H-1‴,Ha-1″,Hb-1″,3″-OH),4.18 (3H,s,5-OCH3),4.12 (1H,br s,6‴-OH),3.73 (1H,t,J=4.9 Hz,H-2″),3.40 (2H,overlap,H-6‴),3.19~3.08 (3H,m,H-3‴,4‴,5‴),3.01 (1H,t,J=8.3 Hz,H-2‴),1.26 (3H,s,3″-CH3),1.24 (3H,s,3″-CH3);13C-NMR(DMSO-d6,100 MHz)δ:159.9 (C-2),149.1 (C-7),146.6 (C-2′),144.3 (C-5),143.1 (C-9),139.8 (C-4),126.6 (C-8),114.8 (C-6),112.8 (C-3),107.1 (C-10),105.9 (C-3′),105.2 (C-1‴),86.0 (C-2″),76.8 (C-5‴),76.6 (C-3‴),74.4 (C-1″),74.2 (C-2‴),71.4 (C-3″),70.0 (C-4‴),61.0 (C-6‴),61.0 (5-OCH3),26.6 (3″-CH3),25.3 (3″-CH3)。NMR数据与文献报道一致[41],故鉴定化合物37为仲-O-β-D-吡喃葡萄糖基白当归素(sec-O-β-D-glucopyranosyl-byakangelicin)。

化合物38:白色固体(甲醇);mp 944 ℃;ESI-MSm/z519 [M+Na]+;1H-NMR (DMSO-d6,400 MHz)δ:8.16 (1H,d,J=9.6 Hz,H-4),8.10 (1H,d,J=2.2 Hz,H-2′),7.35 (1H,d,J=2.2 Hz,H-3′),6.35 (1H,d,J=9.6 Hz,H-3),4.86 (2H,br s,3‴-OH,4‴-OH),4.70 (1H,d,J=7.8 Hz,H-1‴),4.66 (1H,dd,J=10.9,2.8 Hz,Ha-1″),4.37 (1H,d,J=10.9,6.2 Hz,Hb-1″),4.56 (1H,d,J=3.4 Hz,2‴-OH),4.34 (1H,s,2″-OH),4.36 (1H,s,6‴-OH),4.17 (3H,s,5-OCH3),3.94 (1H,dd,J=6.2,2.8 Hz,H-2″),3.69 (1H,dd,J=11.5,2.7 Hz,Ha-6‴),3.44 (1H,m,Hb-6‴),3.25~3.07 (3H,m,H-3‴,4‴,5‴),3.01 (1H,m,H-2‴),1.23 (3H,s,3″-CH3),1.11 (3H,3″-CH3);13C-NMR (DMSO-d6,100 MHz)δ:159.8 (C-2),149.7 (C-7),146.4 (C-2′),144.3 (C-5),143.2 (C-9),139.9 (C-4),126.3 (C-8),114.6 (C-6),112.7 (C-3),107.1 (C-10),105.9 (C-3′),102.0 (C-1‴),82.9 (C-3″),76.9 (C-1″),76.7 (C-2″),75.3 (C-3‴),75.6 (C-5‴),73.9 (C-2‴),70.2 (C-4‴),61.6 (C-6‴),60.7 (5-OCH3),27.4 (3″-CH3),25.5 (3″-CH3)。NMR数据与文献报道一致[41],故鉴定化合物38为叔-O-β-D-吡喃葡萄糖基白当归素(tert-O-β-D-glucopyranosyl-byakangelicin)。

化合物39:白色粉末(乙腈-水);mp 614 ℃;ESI-MSm/z425 [M+H]+。1H-NMR (400 MHz,DMSO-d6)δ:8.03 (1H,d,J=9.5 Hz,H-4),7.69 (1H,s,H-5),6.93 (1H,s,H-8),6.27 (1H,d,J=9.5 Hz,H-3),5.25 (1H,d,J=6.6 Hz,H-3′),5.24 (1H,s,3′-OH),5.01 (1H,s,2″-OH),4.93 (1H,s,3″-OH),4.88 (1H,s,4″-OH),4.53 (1H,d,J=7.1 Hz,H-1″),3.36 (2H,overlap,H-6),3.16 (1H,s,H-3″),3.06 (1H,m,H-4),3.05 (1H,m,H-5),2.88 (1H,m,H-2′),1.47 (6H,s,3″-CH3);13C-NMR (100 MHz,DMSO-d6)δ:162.2 (C-7),160.3 (C-2),156.0 (C-8a),144.8 (C-4),128.5 (C-6),125.6 (C-5),112.7 (C-4a),111.7 (C-3),97.6 (C-1″),97.2 (C-8),91.8 (C-2′),77.4 (C-3′),76.8 (C-3″),76.6 (C-5″),73.3 (C-2″),70.0 (C-4″),69.7 (C-3′),60.7 (C-6″),24.5 (3″-CH3),22.7 (3″-CH3)。NMR数据与文献报道一致[42],故鉴定化合物39为(2′R,3′S)-3′-羟基紫花前胡苷[(2′R,3′S)-3′-hydroxynodakenin]。

化合物40:油状物(丙酮);ESI-MSm/z387 [M+Na]+;1H-NMR (DMSO-d6,400 MHz)δ:8.15 (1H,d,J=9.6 Hz,H-4),8.11 (1H,d,J=2.2 Hz,H-2′),7.69 (1 H,s,H-5),7.09 (1H,d,J=2.2 Hz,H-3′),6.44 (1H,d,J=9.6 Hz,H-3),5.62 (1H,d,J=7.5 Hz,H-1″),5.53 (1H,d,J=5.0 Hz,4″-OH),5.15 (1H,d,J=3.6 Hz,3″-OH),5.03 (1H,br s,2″-OH),4.34 (1H,t,J=5.6 Hz,Ha-6″),3.53 (1H,dd,J=11.6,4.9 Hz,Hb-6′),3.39 (2H,overlap,H-3″,5″),3.21 (2H,br s,H-2″,4″);13C-NMR (DMSO-d6,100 MHz)δ:160.1 (C-2),148.0 (C-2′),146.3 (C-7),145.4 (C-4),142.3 (C-9),128.9 (C-8),126.1 (C-6),116.5 (C-10),114.4 (C-3),114.2 (C-5),107.1 (C-3′),102.1 (C-1″),77.7 (C-5″),76.9 (C-3″),74.1 (C-2″),69.8 (C-4″),60.7(C-6″)。NMR数据与文献报道一致[41],故鉴定化合物40为花椒毒酚-8-O-β-D-吡喃葡萄糖苷(xanthotoxol-8-O-β-D-glucopyranoside)。

化合物41:白色粉末(乙腈-水);mp 236 ℃;ESI-MSm/z290 [M+Na]+。1H-NMR (DMSO-d6,400 MHz)δ:8.36 (1H,s,H-2),8.14 (1H,s,H-8),7.36 (2H,br s,-NH2),5.88 (1H,d,J=6.1 Hz,H-1′),5.47 (1H,d,J=5.7 Hz,2′-OH),5.44 (1H,m,3′-OH),5.21 (1H,d,J=3.8 Hz,4′-OH),4.64 (1H,q,J=5.5 Hz,H-2′),4.15 (1H,d,J=2.8 Hz,H-3′),3.97 (1H,d,J=2.6 Hz,H-4′),3.68 (1H,m,Ha-5′),3.56 (1H,m,Hb-5′);13C-NMR (DMSO-d6,100 MHz)δ:156.2 (C-6),152.4 (C-2),149.0 (C-4),139.9 (C-8),119.3 (C-5),87.9 (C-1′),73.4 (C-2′),70.6 (C-3′),85.9 (C-41′),61.7 (C-5′)。NMR数据与文献报道一致[43],故鉴定化合物41为腺苷(adenosine)。

4 结果与讨论

“都梁丸”为中药复方中典型的药对之一,组方精当。前期工作中,我们首先对组方药味中的白芷[25,38,40-41,44-49]和川芎[50-51]的化学成分进行了详细研究,本文首次把“都梁丸”作为单味药处理,对其物质基础进行了化学成分的研究,分离鉴定了41个化合物,明确了其中的香豆素及香豆素苷类化合物应该来源于白芷,藁本内酯类化合物应该来源于川芎。由于“都梁丸”临床疗效确切,且组方简单,易于采用现代化的分析检测手段对该复方进行研究,从中找出规律,阐明物质基础,研制出现代中药。详细的研究工作嗣后进行。

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ChemicalConstituentsfromCoupletMedicineof“Duliang”Pill

WEI Wei1,WANGLi2a,LIANGLi2a,WANGYuzhen2a,HAILina3,YANGXiuwei1*

(1.StateKeyLaboratoryofNaturalandBiomimeticDrugs,DepartmentofNaturalMedicines,SchoolofPharmaceuticalSciences,PekingUniversity,Beijing100191,China;2.ChangzhiMedicalCollege,Changzhi046000,China;3.BeijingZhendongGuangmingPharmaceuticalResearchInstituteCo.Ltd.,Beijing100120,China)

Objective:To study the chemical constituents of the couplet medicine of "Duliang" Pill.Methods:Repeated column chromatography on silica gel and HPLC were applied to the separation of compounds and their structures were determined by spectroscopic data analyses of MS and NMR.Results:Forty-one compounds were obtained and identified as 5-hydroxymethylfurfural (1),vanillin (2),ferulic acid (3),coniferyl aldehyde (4),xanthotoxin (5),bergaptol (6),falcarindiol (7),senkyunolide H (8),senkyunolide I (9),senkyunolide D (10),4-hydroxy-3-butyl-phthalide (11),senkyunolide A (12),Z-ligustilide (13),3-butylidenephthalide (14),neocnidilide (15),levistolide A (16),β-sitosterol (17),isopimpinellin (18),dibutyl-O-phthalate (19),diisobutyl phthalate (20),chuanxiongdiolide R1(21),atractylenolactam (22),bergapten (23),psoralen (24),isooxypeucedanin (25),phellopterin (26),heraclenol (27),pabularinone (28),anhydrobyakangelicin (29),imperatorin (30),isoimperatorin (31),alloisoimperatorin (32),oxypeucedanin hydrate (33),byakangelicin (34),(-)-marmesinin] (35),(2′S,3′R)-3′-hydroxymarmesinin (36),sec-O-β-D-glucopyranosylbyakangelicin (37),tert-O-β-D-glucopyranosylbyakangelicin (38),(2′R,3′S)-3′-hydroxynodakenin(39),xanthotoxol-8-O-β-D-glucopyranoside (40),and adenosine (41),respectively.Conclusion:Originally,coumarins and coumarin glycosides were from Angelicae Dahuricae Radix,and phthalides were from Chuanxiong Rhizoma.

The couplet medicine;Duliang Pill;Angelicae Dahuricae Radix;Chuanxiong Rhizoma;chemical constituents

国家自然科学基金项目(81473321)

] 杨秀伟,教授,博士生导师,研究方向:中药有效物质基础和药物代谢;Tel:(010)82801569,E-mail:xwyang@bjmu.edu.cn

10.13313/j.issn.1673-4890.2017.7.009

2016-11-13)

*[

a共同第二作者

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