Isolation and Identification of Chemical Constituents of Ethyl Acetate Extraction Layer from Buddleja Officinalis Maxim
-
摘要:
目的 对密蒙花(Buddleja officinalis maxim)乙酸乙酯层的化学成分进行提取分离及结构鉴定。 方法 采用硅胶柱色谱、Spehadex LH-20、HPLC等方法对化学成分进行分离,MS、1H-NMR和13C-NMR鉴定其结构。 结果 分离得到10个化合物,分别鉴定为α-香树脂醇(Ⅰ)、豆甾醇(Ⅱ)、羽扇豆醇(Ⅲ)、环桉烯酮(Ⅳ)、对羟基苯甲酸乙酯(Ⅴ)、羽扇豆醇乙酸酯(Ⅵ)、木犀草素(Ⅶ)、熊果酸(Ⅷ)、α-菠甾醇(Ⅸ)、蒙花苷(Ⅹ)。 结论 Ⅰ,Ⅱ,Ⅲ,Ⅴ,Ⅷ为首次从密蒙花中分离得到。 Abstract:Objective To extract and identify the chemical constituents from ethyl acetate layer of the Buddleja officinalis Maxim. Methods The chemical constituents were isolated by silica gel column chromatography, Spehadex LH-20、HPLC and other methods, and MS、1H-NMR and 13C-NMR were used to identify its structure. Results 10 chemical compounds isolated and identified were α-amyrin (Ⅰ), stigmasterol (Ⅱ), lupeol (Ⅲ), cycloeucalenone (Ⅳ), ethylparaben (Ⅴ), lupeol acetate (Ⅵ), luteolin (Ⅶ), ursolic acid (Ⅷ), α-spinasterol (Ⅸ), linarin (Ⅹ). Conclusion Ⅰ, Ⅱ, Ⅲ, Ⅴ, Ⅷ are the first isolates from the Buddleja officinalis. -
密蒙花又名小锦花、黄饭花、鸡骨头花、染饭花等,为马钱科醉鱼草属植物密蒙花(Buddleja officinalis Maxim)的干燥花蕾和花序[1],在我国主要分布在陕西、甘肃以及西南、中南等地区,生长在海拔200~2800 m的向阳山坡、河边、灌木从中或林缘。该植物最早记载于《开宝本草》[2],具有清热泻火、养肝明目、退翳功效。本文对密蒙花乙酸乙酯层中的化学成分进行系统性研究,为该植物的开发利用奠定了理论基础。
1. 材料与方法
1.1 材料与仪器
1.1.1 实验原料
原料密蒙花采摘于云南昆明,摘掉叶和茎,留下花蕾和花序部分。
1.1.2 实验设备
设备仪器ZF-2型三用紫外分析仪,上海安亭电子仪器厂:VGAUTO Spec-3000型质谱仪,美国Thermo公司;Bruker AM-400,AVANCEⅢ 500 MHz,AVANCE Ⅲ 600 MHz型核磁共振仪,德国,Brucker公司。
1.2 实验方法
方法干燥密蒙花花蕾和花序7.8 kg,用75%乙醇浸提3次,每次72 h,过滤,合并提取液,回收乙醇得到总浸膏。总浸膏用蒸馏水溶解,用等体积乙酸乙酯和正丁醇依次萃取3次,分别得到乙酸乙酯层、正丁醇层和水层,分别浓缩得其浸膏。取乙酸乙酯层浸膏(150 g),经硅胶柱色谱和Spehadex LH-20凝胶柱色谱反复梯度洗脱得到单体化合物,通过MS、1H-NMR和13C-NMR鉴定其结构。
2. 结果
本实验从密蒙花乙酸乙酯层浸膏分离得到10个化合物,鉴定结果如下,结构如图1所示。
化合物Ⅰ:白色针状(氯仿),分子式C30H50O,ESI-MS m/z,426 [M + H]+。1H NMR(500 MHz,CDCl3)δ:5.13(1H,t,J = 3.6 Hz,H-12),3.22(1H,dd,J = 5.2,11.0 Hz,H-3),1.07(3H,s),1.01(3H,s),1.00(3H,s),1.00(3H,d,J = 7.0 Hz),0.95(3H,d, J = 7.0 Hz),0.91(3H,s),0.80(3H,s),0.79(3H,s)。13C NMR(126 MHz,CDCl3)δ:38.8(C-1),27.2(C-2),79.0(C-3),39.2(C-4),55.3(C-5),18.4(C-6),32.9(C-7),40.0(C-8),47.7(C-9),36.9(C-10),23.2(C-11),124.43(C-12),139.59(C-13),42.1(C-14),28.5(C-15),26.6(C-16),33.8(C-17),59.1(C-18),39.7(C-19),39.6(C-20),31.3(C-21),41.5(C-22),28.2(C-23),15.6(C-24),15.7(C-25),16.9(C-26),23.3(C-27),28.8(C-28),17.5(C-29),21.4(C-30)。
化合物Ⅱ:白色粉末状(氯仿),分子式C29H48O,ESI-MS m/z:412 [M + H]+。1H NMR(500 MHz,CDCl3)δ:5.35(1H,m,H-6),5.15(1H,dd,J = 15.0,8.5 Hz,Ha-22),5.02(1H,dd, J = 15.0,8.5 Hz,Ha-23),3.52(1H,m,H-3),1.03(3H,s,H-19),1.01(3H,d, J = 7.0 Hz,H-21),0.85(3H,t,J = 6.5 Hz,H-29),0.82(3H,d,J = 7.0 Hz,H-26),0.80(3H,d,J = 7.0 Hz,H-27),0.70(3H,s,H-18)。13C NMR(126 MHz,CDCl3)δ:37.3(C-1),31.7(C-2),71.8(C-3),42.3(C-4),140.8(C-5),121.7(C-6),31.9(C-7),32.0(C-8),50.2(C-9),36.5(C-10),21.1(C-11),39.7(C-12),42.2(C-13),56.9(C-14),24.4(C-15),28.9(C-16),56.0(C-17),12.0(C-18),19.4(C-19),40.5(C-20),21.2(C-21),138.3(C-22),129.3(C-23),51.2(C-24),32.0(C-25),19.0(C-26),21.1(C-27),25.4(C-28),12.2(C-29)。
化合物Ⅲ:白色无定型粉末(氯仿),分子式C30H50O,ESI-MS m/z:426[M + H]+。1H NMR(500 MHz,CDCl3)δ:3.17(1H,dd,H-3),2.36(1H,m,H-18),2.27(1H,m,H-7),1.66(3H,m,H-30),1.65(1H,m,H-1),1.50~1.70(2H,m,H-2),1.50(1H,m,H-6),1.36(1H,m,H-6),1.36(1H,m,H-7),1.38(1H,m,H-11),1.25(1H,s,H-9),1.23(1H,m,H-11),1.65(1H,s,H-12),1.01(1H,s,H-12),1.64(1H,s,H-13),0.96(1H,m,H-15),1.10(1H,m,H-15),1.47(1H,m,H-16),1.38(1H,m,H-22),1.36(1H,m,H-16),1.35(1H,m,H-19),1.24(1H,s,H-21),1.18(1H,m,H-22),1.02(3H,s,H-26),0.95(3H,s,H-23),0.93(3H,s,H-27),0.89(1H,m,H-1),0.81(3H,s,H-25),0.77(3H,s,H-28),0.74(3H,s,H-24),0.66(1H,d,J = 9.0 Hz,H-5)。13C NMR(126 MHz,CDCl3)δ:38.7(C-1),27.4(C-2),79.0(C-3),38.8(C-4),55.2(C-5),18.3(C-6),34.2(C-7),40.8(C-8),50.4(C-9),37.1(C-10),20.9(C-11),25.1(C-12),38.0(C-13),42.8(C-14),27.4(C-15),35.6(C-16),43.0(C-17),48.0(C-18),48.3(C-19),151.0(C-20),29.8(C-21),40.0(C-22),28.0(C-23),15.4(C-24),16.0(C-25),16.1(C-26),14.5(C-27),18.0(C-28),19.4(C-29),109.3(C-30)。
化合物Ⅳ:黄色粉末状(氯仿),分子式C30H48O,ESI-MS m/z:424[M + H]+。1H NMR(500 MHz,CDCl3)δ:4.72(1H,s,Ha-30),4.67(1H,s,30-Hb),1.02(18H,m,18,21,26,27,28,29-Me)。13C NMR(126 MHz,CDCl3)δ:32.8(C-1),41.4(C-2),213.5(C-3),50.8(C-4),46.2(C-5),25.3(C-6),28.2(C-7),47.8(C-8),24.4(C-9),29.1(C-10),25.9(C-11),35.1(C-12),45.3(C-13),48.8(C-14),32.7(C-15),27.6(C-16),53.2(C-17),18.0(C-18),27.3(C-19),36.2(C-20),18.8(C-21),34.9(C-22),31.3(C-23),56.4(C-24),33.0(C-25),21.1(C-26),22.5(C-27),19.2(C-28),10.7(C-29),105.9(C-30)。
化合物Ⅴ:无色结晶(氯仿),分子式C9H10O3,ESI-MS m/z:166[M + H]+。1H NMR(500 MHz,CDCl3)δ:7.47(1H,d,J = 8.6Hz),6.75(2H,d,J = 8.6Hz),4.78(1H,q,J = 7.1 Hz)。13C NMR(126MHz,CDCl3,)δ:166.9(-COOH),160.5(C-4),131.9(C-2,6),115.9(C-3,5),60.9(-OCH2),14.3(-CH3)。
化合物Ⅵ:白色粉末状(氯仿),分子式C32H52O2,ESI-MS m/z:468 [M + H]+。1H NMR(500 MHz,CDCl3)δ:0.79(3H,s,H-28),0.84(3H,s,H-24),0.85(3H,8,H-23),0.86(3H,s,H-25),0.94(3H,s,H-27),1.03(3H,s,H-26),1.68(3H,s,30-CH3),2.03(3H,8,O = CCH3),4.47(1H,m,H-3)。13C NMR(126 MHz,CDCl3)δ:14.5(C-27),16.0(C-26),16.1(C-25),16.4(C-24),18.0(C-28),18.2(C-6),19.3(C-29),20.9(C-11),21.2(C-31),23.7(C-2),25.1(C-12),27.4(C-15),27.9(C-23),29.8(C-21),34.2(C-7),35.5(C-16),37.1(C-10),37.8(C-4),38.4(C-1),40.0(C-22),40.8(C-8),42.8(C-14),43.0(C-17),48.3(C-18),48.0(C-19),50.3(C-9),55.4(C-5),80.99(C-3),109.3(C-30),150.9(C-20),171.9(C-32)。
化合物Ⅶ:黄色粉末状(甲醇),分子式C15H10O6,ESI-MS m/z:286 [M + H]+。1H NMR(500 MHz,MeOD)δ:6.68(1H,s,H-3),6.20(1H,d,J = 2.4 Hz,H-6),6.45(1H,d,J = 2.4 Hz,H-8),7.40(1H,d,J = 2.4 Hz,H-2′),6.90(1H,d,J = 8.4 Hz,H-5′),7.43(1H,dd,J = 8.4,2.4 Hz,H-6′)。13C NMR(126 MHz,MeOD)δ:166.3(C-2),104.1(C-3),183.1(C-4),163.9(C-5),100.2(C-6),166.5(C-7),94.3(C-8),159.7(C-9),103.3(C-10),120.4(C-1′),114.8(C-2′),147.2(C-3′),150.1(C-4′),116.4(C-5′),123.9(C-6′)。
化合物Ⅷ:白色粉末状(甲醇),分子式C30H48O3,ESI-MS m/z:456 [M + H]+。1H NMR(500 MHz,MeOD)δ:5.37(1H,brs,H-12),3.19(1H,m,H-3),2.78(1H,m,H-18),1.11(3H,s,H-27),0.89(3H,s,H-25),0.88(3H,s,H-26),0.75(3H,s,H-23),0.74(3H,s,H-24)。13C-NMR(126 MHz,MeOD)δ:38.8(C-1),27.4(C-2),79.5(C-3),38.8(C-4),55.2(C-5),18.3(C-6),32.2(C-7),39.8(C-8),47.4(C-9),37.1(C-10),22.9(C-11),126.6(C-12),139.3(C-13),41.8(C-14),27.7(C-15),23.6(C-16),46.2(C-17),41.0(C-18),45.3(C-19),30.2(C-20),33.8(C-21),32.9(C-22),28.3(C-23),15.4(C-24),15.3.(C-25),17.1(C-26),25.5(C-27),180.8(C-28),33.7(C-29),23.9(C-30)。
化合物Ⅸ:白色粉末状(氯仿),分子式C29H48O,ESI-MS m/z:412 [M + H]+。1H NMR(500 MHz,CDCl3)δ:5.17(3H,m,H-7,22,23),3.69(1H,m,3-OH),1.03(3H,d,J = 6.6 Hz,H-21),0.81(3H×4,H-19,26,27,29),0.56(3H,s,H-18)。13C NMR(126 MHz,CDCl3)δ:37.1(C-1),31.4(C-2),71.5(C-3),38.8(C-4),40.2(C-5),29.3(C-6),117.2(C-7),139.8(C-8),49.4(C-9),34.6(C-10),21.9(C-11),39.6(C-12),43.3(C-13),55.8(C-14),28.7(C-15),23.6(C-16),55.2(C-17),12.0(C-18),13.3(C-19),40.2(C-20),21.8(C-21),138.9(C-22),129.3(C-23),51.4(C-24),31.3.(C-25),21.1(C-26),19.5(C-27),25.8(C-28),12.7(C-29)。
化合物Ⅹ:淡黄色粉末(甲醇),分子式C28H32O14,ESI-MS m/z:591 [M + H]+。1H NMR(500 MHz,MeOD)δ:1.09(3H,d,J = 6.0 Hz,H-6),3.16(1H,dd,J = 9.0,9.0 Hz,H-4″),3.16(1H,dd,J = 9.0,9.0 Hz,H-4),3.28(1H,dd,J = 7.0,9.0 Hz,H-2″),3.32(1H,dd,J = 9.0,9.0 Hz,H-3″),3.41~3.43(1H,m,H-5),3.47(1H,dd, J = 4.5,12.0 Hz,H-6″),3.48(1H,dd,J = 3.0,9.0 Hz,H-3),3.61~3.63(1H,m,H-5″),3.68(1H,br d,J = 3.0 Hz,H-2),3.87(3H,s,4'-OCH3),3.88(1H,br d,J = 12.0 Hz,H-6″),4.56(1H,br s,H-1),5.07(1H,d,J = 7.0 Hz,H-1″),6.46(1H,br s,H-6),6.80(1H,br s,H-8),6.95(1H,s,H-3),7.15(2H,d,J = 8.5 Hz,H-3',5'),8.05(2H,d,J = 8.5 Hz,H-2',6'),12.92(1H,br s,5-OH)。13C NMR(126 MHz,MeOD)δ:183.9(C-4),166.9(C-2),166.0(C-7),163.4(C-4'),162.1(C-5),159.9(C-9),129.4(C-2',6'),123.6(C-1'),117.7(C-3',5'),105.4(C-10),103.8(C-3),100.5(Rha-C-1),99.9(Glc-C-1),99.6(C-6),94.8(C-8),76.2(Glc-C-3),75.7(GlcC-5),73.1(Glc-C-2),72.0(Rha-C-4),70.7(Glc-C-4),70.3(Rha-C-2),69.6(Rha-C-3),68.3(Rha-C-5),66.1(Glc-C-6),55.5(OCH3),17.7(Rha-C-6)。
3. 讨论
化合物Ⅰ的数据与文献报道[3]的α-香树脂醇基本一致;化合物Ⅱ与文献报道[4]的豆甾醇的数据基本一致;化合物Ⅲ与文献报道[5]的羽扇豆醇的数据基本一致;化合物Ⅳ与文献报道[6]的环桉烯酮的数据基本一致;化合物Ⅴ与文献报道[7]的对羟基苯甲酸乙酯数据基本一致;化合物Ⅵ与文献报道[8]的羽扇豆醇乙酸酯数据基本一致;化合物Ⅶ与文献报道[9]的木犀草素数据基本一致;化合物Ⅷ与文献[10]报道的熊果酸数据基本一致;化合物Ⅸ与文献报道[11]的α-菠甾醇数据基本一致;化合物Ⅹ与文献[12]报道的蒙花苷数据基本一致。
密蒙花乙醇提取物的乙酸乙酯层萃取部分经过硅胶柱层析和Spehadex LH-20凝胶柱色谱,再通过波普数据分析与文献数据对比,鉴定了10个单体化合物。它们分别为α-香树脂醇、豆甾醇、羽扇豆醇、环桉烯酮、对羟基苯甲酸乙酯、羽扇豆醇乙酸酯、木犀草素、熊果酸、α-菠甾醇、蒙花苷。其中α-香树脂醇、豆甾醇、羽扇豆醇、对羟基苯甲酸乙酯、熊果酸为该植物中首次分离。密蒙花单体化合物的分离,为以后的药理活性研究提供理论依据,为以后密蒙花的研究、开发和利用奠定基础。
-
[1] 郭雷,朱文成,刘超. 密蒙花化学成分及生物活性研究进展[J]. 食品研究与开发,2012,33(7):222-225. doi: 10.3969/j.issn.1005-6521.2012.07.063 [2] 杨东方,胡云飞,丁倩倩,等. 密蒙花的化学成分及质量控制方法研究进展[J]. 中国现代应用药学,2019,36(22):2870-2875. [3] Yan Huang,Xu Wu,Jianwen Wen,et al. A Study on the Chemical Constituents of Pileasinofasciata C. J. Chen[J]. Medicinal Plant,2017,8(3):1-4. [4] 薛鹏辉,段静诗,丁丽琴,等. 鹅不食草中甾体及酚类化学成分研究[J]. 中国药物化学杂志,2020,30(6):340-346. [5] 孙志青,刘立云,孙岩,等. 血芙蓉化学成分的研究[J]. 中成药,2020,42(4):932-938. doi: 10.3969/j.issn.1001-1528.2020.04.021 [6] 吴贵辉,陈艳,郑黎花,等. 黔产杜仲化学成分研究[J]. 中药材,2015,38(5):980-984. [7] 张鹏耀,舒任庚,严喜鸾,等. 山蜡梅化学成分研究[J]. 中国药学杂志,2017,52(18):1594-1596. [8] Chen Hui,Li Yu Jie,Sun Yan Jun,et al. Antihyperlipidemic glycosides from the root bark of Lyciumchinense.[J]. Natural Product Research,2019,33(18):2655-2661. doi: 10.1080/14786419.2018.1466125 [9] 何丽丽,姚彩云,闫炳雄,等. 壮药山风中的黄酮类化学成分研究[J]. 中国现代中药,2019,21(8):1016-1020. [10] 孙彦君,陈豪杰,高美玲,等. 小驳骨中萜类化学成分研究[J]. 中草药,2020,51(2):293-298. [11] 刘宝,胡飞龙,雷福厚,等. 灰色紫金牛化学成分研究[J]. 中药材,2019,42(3):560-562. [12] 郑畅,阮静雅,瞿璐,等. 密蒙花中黄酮类成分的分离与鉴定[J]. 中国药物化学杂志,2018,28(1):52-57. -