STUDIES ON THE 13C-NMR SPECTRA OF ALTERNATING COPOLYMERS OF CONJUGATED DIENES WITH METHYL A
褐煤黄腐酸钾制黄腐酸的分子结构表征
褐煤黄腐酸钾制黄腐酸的分子结构表征阳虹;张玉贵;李永生;江林华【摘要】为了进一步了解黄腐酸(FA)的结构和性质,为拓宽黄腐酸在工农业领域中的应用范围打下基础,用傅里叶红外光谱、紫外-可见光光谱和固体13C交叉极化/魔角旋转核磁共振(13C-CP/MAS NMR)对FA的结构进行表征.结果表明,FA的分子结构由芳香核和核外侧链构成,其中芳香碳约占54％,芳香核主要由4个芳香环构成.脂肪碳约占46％,脂肪碳的存在形式主要是含氧的五元或六元脂肪环.FA中含氧基团的含量为32.3％,主要由醌基和羟基构成,还有部分羧基.【期刊名称】《河南理工大学学报（自然科学版）》【年(卷),期】2014(033)004【总页数】4页(P539-542)【关键词】黄腐酸;紫外光谱;红外光谱;核磁共振【作者】阳虹;张玉贵;李永生;江林华【作者单位】河南理工大学物理化学学院,河南焦作454000;河南省瓦斯地质与瓦斯治理重点实验室-省部共建国家重点实验室培育基地(河南理工大学),河南焦作454000;河南省瓦斯地质与瓦斯治理重点实验室-省部共建国家重点实验室培育基地(河南理工大学),河南焦作454000;昊华骏化集团股份有限公司,河南驻马店463000;河南省瓦斯地质与瓦斯治理重点实验室-省部共建国家重点实验室培育基地(河南理工大学),河南焦作454000【正文语种】中文【中图分类】O657.610 引言随着我国经济的发展，能源瓶颈问题日益突出，因此，开发新能源及能源的多元化利用是能源发展的必然趋势.FA因其在水中、土壤、泥炭、风化煤、城市废弃物中广泛存在而引起学者们的注意，对于FA的提取、结构表征和利用，学者们做了大量的研究工作，如Gennaro等在植物秸秆的堆肥过程中研究了腐植酸的结构性质，认为堆肥在一定的条件下可以获得和土壤腐植酸相似的结构，可以作为土壤的改良剂［1］.Roberto等研究了两种黄腐酸的结构，运用多种分析方法，通过桥键，疏水作用等，解释了黄腐酸在溶液中为什么有形成聚合体的现象［2］.Etelvino等曾运用核磁共振的方法研究认为土壤腐植酸中含有高度稠和的芳碳和大量的不稳定的含氧结构［3］.Jarafshanj等通过离子强度、腐植酸浓度和pH值等方面详细地研究了标准腐植酸物质的三维荧光效应［4］.本文着重研究了从褐煤黄腐酸钾制取的黄腐酸的分子结构，为拓宽腐植酸的应用打下一定基础.1 实验1.1 黄腐酸的提取将732型阳离子交换树脂预处理为强酸型［5］，取购置于双龙腐植酸公司生产的黄腐酸钾(由褐煤硝酸氧解所制)，按黄腐酸钾、水和树脂的质量比例为1∶10∶20，混合搅拌进行离子交换，静置过夜(保证液体显酸性)，过滤，将溶液在烘箱中60℃下烘干，即得黄腐酸.1.2 官能团含量的测定总酸性基测定用碱溶氯化钡沉淀电位滴定法，羧基含量测定用羧基微量快速测定法［6］，羟基的含量为总酸性基含量减去羧基含量，所用实验试剂均为分析纯.结果如表1所示.表1 官能团含量Tab.1 Content of functional groups mmol/g?1.3 傅里叶红外光谱取干燥的FA样品和KBr按1∶150的比例压片，用德国Vertex 70傅里叶变换红外光谱仪记录4 000～400 cm-1范围内的红外光谱.分辨率为4 cm-1.1.4 紫外-可见光谱取干燥的 FA样品50 mg，溶解在0.1 mol/L的NaHCO3溶液中(加少量的NaOH使 FA溶解)，并定容于1 L的棕色容量瓶配制成50 mg/L的溶液，在TU-1900紫外-可见分光光度计上扫描，范围为220～600 nm，记录其吸光度的变化.1.5 13C-CP/MAS NMR 的测定测定13C-CP/MAS NMR的仪器为德国Bruker Avance AV 400超导核磁共振仪，转子为4 mm，转速 8 000 r/min，频率 400 MHz，谱宽 30 241.936 Hz，采样时间 AQ 0.033 910 3 s，脉宽 2 000 u/s，延迟时间D为13 s，扫描次数Ns为2 048次.2 结果与讨论2.1 黄腐酸的红外光谱分析把FA的吸收分为4个大的吸收带:3 000～3 600 cm-1为羟基吸收带，吸收带最明显的特征是在3 400 cm-1处有一大宽峰，表明FA含有大量的羟基，氢键缔合现象较明显.2 800～3 000 cm-1之间的特征吸收峰为脂肪结构吸收带，图1中较为明显的有2 850 cm-1附近的CH2对称伸缩振动、2 930 cm-1附近的CH2非对称伸缩振动，其余的则较难观测到.1 000～1 800 cm-1主要为含氧官能团吸收带，其中1 720 cm-1左右有尖锐的羧基的伸缩振动峰，表明羧基很少发生缔合;FA在1 609 cm-1处具有吸收峰，有人认为是与氢缔合的醌基［1］，本文认为这是芳香共轭双键的吸收峰，是分子芳香性和缩聚度的指标;在1 512，1 460 cm-1处尖锐的吸收峰是芳香环的骨架振动.1 420 cm-1处的为羟基的振动峰;1213 cm-1处的宽峰认为是两个峰的合峰，分别为1 213 cm-1，1 175 cm-1和1 213 cm-1处的可归于酚醚、或醇C—O伸展和C—OH振动;1175 cm-1的可归于甲氧基、醇醚的振动;FA在1 030 cm-1处有峰，可能是饱和醇的C—O伸缩振动或者是灰分所引起.700～900 cm-1为芳香结构吸收带，其中863 cm-1处的峰可能是取代芳环CH面外变形振动，表明芳环周围有被取代基取代的现象.由红外图谱可知，FA中含有大量的含氧官能团，芳环周围有一定的含氧取代基，脂肪侧链以亚甲基结构为主.2.2 黄腐酸的紫外-可见光谱分析紫外-可见光谱法，是基于分子内电子跃迁产生吸收光谱的一种分析方法，紫外-可见光谱中吸收峰的位置和强度与分子的不饱和键及取代基密切相关.从图2中可以看出，FA在整个波长范围内的吸收光谱是吸光度随着波长的增大而减小的混合光谱.在280 nm处有一吸收平台，它是由苯环吸收造成的［7］;230 nm处有一最大吸收峰，表明分子中可能含有两个双键的共轭体系;鉴于Tsutsuki 等用硼氢化钠和连二亚硫酸钠还原各种土壤腐植酸，发现紫外和可见光吸收主要是醌基、醛基和非醌羰基的贡献［8］.因此，认为230 nm处的吸收峰可能来源于腐植酸中大量存在的不饱和醛基、醌基等.成绍鑫等也用此还原方法研究了煤炭腐植酸和硝基腐殖酸的光谱特征，发现煤炭腐植酸和硝基腐殖酸用NaBH4还原后可见光区域吸收峰明显降低，硝基腐殖酸的光谱图形和Tsutsuki等提出的RP型土壤腐植酸的单环共轭醌结构很相似［9］.这也和前文红外光谱中含有较多的含氧官能团相一致.2.3 13C-CP/MAS NMR 分析图3为FA的13C-CP/MAS NMR谱图，它可以划分为3个区域，分别为脂肪碳(δ0-90)，芳香碳(δ90-165)，羰基碳(δ190-210).利用 nuts2008 对这3个区域的谱图进行分峰拟合，由图3可以看到实验谱图和拟合谱图吻合得比较好，说明本文对FA的13C-CP/MAS NMR谱图进行的分峰是合适的.根据FA样品中的峰位及其相对含量，可计算出FA的结构参数，见表2.由于在FA结构中存在不同类型的脂肪碳，Fal为FA中脂肪碳(总含量为44%).Fal 为和之和.为FA中CH3的相对含量为FA中季碳，CH，CH2的相对含量为FA中和氧相接的脂碳.由表2可知，FA中的比值为13∶3∶28.由图3 知，FA 有δ30和δ50 两个明显的突出峰，分别为的含量.亚甲基和甲基的比例为4∶1，氧接脂碳和亚甲基的比例为2∶1，FA的脂肪碳中以氧接脂碳和亚甲基为主，甲基的含量很少，可以推测FA中的脂肪碳主要以含氧的五元、六元环脂肪环或者长链含氧脂肪侧链构成.表2 FA的13C-CP/MAS NMR结构参数［10］Tab.2 13C-CP/MAS NMR parameters of FA samples注:fa为芳香总碳为羰基碳;Fa′为芳环碳;为质子化碳为非质子化碳;酚或脂碳;烷基化芳碳;为桥碳;Fal为脂肪总碳;为与氧相连的脂碳;Xb为桥头芳碳在芳碳原子中所占的百分数?FA中芳香碳的含量为54%分别为芳环的质子化碳和非质子化碳的比例约为1∶3.5，说明芳香核上有77%的碳被取代.在非质子化碳中为3∶8∶30.非质子化碳中70%以桥键的形式存在，说明FA中的芳香核由较多的芳环构成为3∶8，结合上述的氧接脂碳和亚甲基的比例为2∶1，亚甲基和甲基的比例为4∶1，更可以说明在芳香环上面的以亚甲基为主，酚羟基和醚氧连碳为辅，亚甲基上连接氧构成脂肪环.与上述脂肪碳的预测存在形式相一致.在δ160处的轻微小突出峰，也是FA中含有少量氧接脂碳的一种证明，根据表2和紫外光谱的结论，这些氧接脂碳可能为醌基或羟基.芳香桥碳与周碳之比)指桥头芳碳在芳碳原子中所占的百分数［11］，计算得知芳香核主要以4个芳环构成.FA中羰基碳的含量非常少，仅为1.12%.由表1和红外图谱也可以知道，FA中的羧基含量很少，结合紫外可见光谱中的数据，可以推测羰基碳应主要由醌基构成.FA中的含氧官能团［12］(+f)的含量为32.3%.3 结论通过对傅里叶红外光谱、紫外-可见光谱研究，得出FA中含有大量的含氧官能团，芳环周围有一定的含氧取代基，脂肪侧链以亚甲基结构为主.推测出FA含有一些像醌基和醛基一样的含氧共轭体.通过对13C-CP/MAS NMR的研究，得出FA的分子由芳香核和核外侧链构成，其中芳香碳占54%，芳香核主要由4个芳香环构成;脂肪碳占44%，脂肪碳的存在形式主要是含氧的五元或六元脂肪环，FA含氧基团的含量为32.3%.参考文献:［1］ BRUNETTI G，SOLER R P，MATATRESE position and Structural Characteristics of Humified Fractions during the Co-composting Process of Spent Mushroom Substrate and Wheat Straw ［J］.Agric Food Chem，2009，57:10859-10865.［2］ ROBERTOB，MARTA F，GUSTAVO G plementary Multianalytical Approach To Study the Distinctive Structural Features ofthe Main Humic Fractions in Solution:Gray Humic Acid，Brown Humic Acid，and Fulvic Acid［J］.Agric food chem，2009.57.3266-3272.［3］ ETELVINO H N，EDUATDO R D.TITO J B.Studies of the Compositions of Humic Acids from Amazonian Dark Earth Soils［J］.Environ Sci Technol，2007，41(2):400-405.［4］ JARAFSHAN J，MOBED S I，HEMMINGSEN X.FluorescenceCharacterization of IHSS Humic.Substances:Total Luminescence Spectra with Absorbance Correction［J］.Environ Sci Technol，1996，30(10):3061-3065.［5］焦元刚.从风化煤中提取黄腐酸［D］.北京:北京交通大学，2006.［6］李善祥.腐植酸产品分析及标准［M］.北京:化学工业出版社，2007:78-84. ［7］徐栋，冯科，昊峰.天然水体底质中腐植酸的光谱表征［J］.分析科学学报，2003，19(6):499-502.［8］成绍鑫.腐植酸类物质概论［M］.北京:化学工业出版社，2006:104.［9］成绍鑫，孙淑和，吴奇虎.腐植酸中羰基、醌基对紫外可见光谱的影响［J］.江西腐植酸，1982(2):1.［10］ MARK S S，RONALD J P，DAVID M G.13C Solid-state NMR of Argonne Premium Coals［J］.Energy Fuels，1989，3，187-193.［11］贾建波，曾凡桂，孙蓓蕾.神东2#煤镜质组大分子结构模型13C-NMR谱的构建与修正［J］.燃料化学学报，2011，39(9)，652-656.［12］罗陨飞，李文华，陈亚飞.中低变质程度煤显微组分结构的13C-NMR研究［J］.燃料化学学报，2005，33(5):540-543.。
3版仪器分析第5节-13C核磁共振波谱课件
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第十九章 核磁共振波谱
分析法
nuclear magnetic resonance spectroscopy; NMR
第五节 13C核磁共振谱简介
一、 概述 generalization 二、 化学位移 chemical shift 三、 偶合与弛豫 coupling and relaxation 四、 13CNMR谱图 13C NMR spectrograph
H splitting
I=
1 2
C splitting
1
I= 2
( 19F：300ppm； 31P：700ppm；)
H0
（3）13C-13C偶合的几率很小；13C天然丰度1.1%；
（4）13C-H偶合可消除，谱图简化。
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PFT-NMR
MAGNET MAGNET
R.F.transmitter
R-F receiver and detector
Sweep generator
Recorder
(1) Magnet 25 1 ℃ (2) Sweep generator 3~10mG/min 全程~0.2Gauss (3) （射频发生器 ） R–F transmitter (4) （射频接收器和检测器） R–F receiver and detector (5) （样品支架 , 探头） Sample holder ,probe (6) （记录仪） Recorder
C13-NMR
13C NMR谱图
1.C7H14O
Saturated carbon - sp3 electronegativity effects
40 - 80
35 - 80 25 - 65
Unsaturated carbon - sp2
C C=C
C
Alkyne carbons - sp
65 - 90
100 - 150 110 - 175
Aromatic ring carbons C=O C=O 200
化学位移规律：烷烃
碳数n >4 端甲基 C=13-14
C>CH> CH2 >CH3
邻碳上取代基增多C 越大
取代烷烃:
H 3C 13.9
C H2 22.8
C H2 34.7
C H2
C H3
化学位移规律：烯烃
C C
C=100-150（成对出现） 端碳=CH2 110；邻碳上取代基增多C越大：
香豆精反转门去偶与宽带去偶谱20015010050rch30range830155520604080358025656590100150110175155185185220saturatedcarbonsp3noelectronegativityeffectsrch2rr3chr4ccocclcbrsaturatedcarbonsp3electronegativityeffectsunsaturatedcarbonsp2ccccalkynecarbonssparomaticringcarbonscoco200acidsamidesestersanhydridesaldehydesketones150100500correlationchartfor13cchemicalshiftsppm应该熟记的c13nmr位移位移应该熟记的approximate13cchemicalshiftrangesforselectedtypesofcarbonppmrch3r2ch2r3chcicbrcclcnco830155520600402565358030654080cccccn6590100150110140110175oorcorrcohorcnh2oorchrcr1551851551851852201propanolhoch2ch2ch3cbaprotondecoupled200150100500protondecoupled13cspectrumof1propanol225mhz22dimethylbutane22bromocyclohexanecyclohexanoltoluenecyclohexenecyclohexanone12dichlorobenzene12baabcclccl13dichlorobenzene13accldbclsolventad影响化学位移因素杂化轨道电子短失正碳位移250330ppm孤电子对有未享用的孤电子对该碳向低场移约50ppm位移增加50ppm影响化学位移因素电负性共轭效应氢键会导致碳电子密度降低构型因素顺式一般在高场低位移弛豫13c的弛豫要比1h慢很多1h的t常在1011秒而13c常大于1s
CNMR
40
20
0
Chemical shift (, ppm)
Comparison Between 1H and 13C NMR
Similarity Both give us information about the number of chemically nonequivalent nuclei (nonequivalent hydrogens or nonequivalent carbons) Both give us information about the environment of the nuclei (hybridization state, attached atoms, etc.)
13C
Proton Spectrum
1H
ClCH2CH2CH2CH2CH3
ClCH2
CH3
10.0
9.0
8.0
7.0
6.0
5.0
4.0
3.0
2.0
1.0
0
Chemical shift (, ppm)
Carbon Spectrum
13C
ClCH2CH2CH2CH2CH3 a separate, distinct peak appears for each of the 5 carbons
CH3
7 carbons give 7 signals, but intensities are not equal.
OH
200
180 160 140 120 100 80
Chemical shift (, ppm)
60
40
20
0
13C-2D
Coupling
Euodionosides A–G Megastigmane glucosides from leaves of Euodia meliaefolia
Euodionosides A–G:Megastigmane glucosides fromleaves of Euodia meliaefoliaMiwako Yamamoto a ,Takeyuki Akita a ,Yuka Koyama a ,Etsuko Sueyoshi a ,Katsuyoshi Matsunami a ,Hideaki Otsuka a,*,Takakazu Shinzato b ,Atsushi Takashima c ,Mitsunori Aramoto d ,Yoshio Takeda eaGraduate School of Biomedical Sciences,Hiroshima University,1-2-3Kasumi,Minami-ku,Hiroshima 734-8553,Japan bSubtropical Field Science Center,Faculty of Agriculture,University of the Ryukyus,1Sembaru,Nishihara-cho,Nakagami-gun,Okinawa 903-0213,JapancYona Field,Subtropical Field Science Center,Faculty of Agriculture,University of the Ryukyus,685Aza Yona,Kunigami-son,Kunigami-gun,Okinawa 905-1427,JapandIriomote Station,Tropical Biosphere Research Center,University of the Ryukyus,870Aza Uehara,Taketomi-cho,Yaeyama-gun,Okinawa 907-1541,JapaneFaculty of Integrated Arts and Sciences,The University of Tokushima,1Minamijosanjima-cho,Tokushima 770-8502,JapanReceived 9August 2007;received in revised form 28December 2007Available online 11March 2008AbstractFrom a 1-BuOH-soluble fraction of the MeOH extract of leaves of Euodia meliaefolia ,collected in Okinawa,seven megastigmane glucosides,named euodionosides A–G,were isolated together with three known megastgmane glucosides,and two aliphatic and three phenolic compounds.Their structures were elucidated through a combination of spectroscopic analyses and application of the modified Mosher’s method.Ó2008Elsevier Ltd.All rights reserved.Keywords:Euodia meliaefolia ;Rutaceae;Euodionoside;Megastigmane glucoside;Modified Mosher’s method1.IntroductionThe fruits of Evodia rutaecaepa (Rutaceae)are a well known crude drug included in the Japanese Pharmaco-poeia XV and contain characteristic indole alkaloids,i.e.rutaecarpine and evodiamine.However,in reliable plant name databases (IPNI ),such as the Index Kewensis,Aus-tralian Plant Name Index and Gray Card Index,the genus name E v odia is not listed under the Rutaceae.Instead,many Euodia species appear in the databases as Rutaceous plants.Probably,some previous investigators made a mis-take due to the resemblance of the letters u and v.Thereaf-ter,even an isolated compound was erroneously named evodiamine.A closely related species,E.meliaefolia ,is a tall deciduous tree of about 15m in height and found in southern Kyushu through Okinawa,Taiwan and China (Hatusima,1975).Currently no chemical and medicinal investigations of this plant have been reported.In a continuing study on Okina-wan plants,the chemical constituents of E .meliaefolia ,collected in Okinawa,were investigated.From a 1-BuOH-soluble fraction of the MeOH extract of leaves of E .meliaefolia ,10megastigmane glucosides (1–10)were isolated together with two known aliphatic [(Z )-3-hexenyl b -D -glucopyranoside (11)(Mizutani et al.,1988)and 3,7-dimethylocte-1-en-3,6,7-triol 6-O -b -D -gluco-pyranoside (12)(Manns,1995)]and three phenolic [(+)-catechin (13)(Nay et al.,2001),syringin (14)DellaGreca0031-9422/$-see front matter Ó2008Elsevier Ltd.All rights reserved.doi:10.1016/j.phytochem.2008.01.020*Corresponding author.Tel./fax:+81822575335.E-mail address:hotsuka@hiroshima-u.ac.jp (H.Otsuka)./locate/phytochemAvailable online at Phytochemistry 69(2008)1586–1596PHYTOCHEMISTRYet al.,1998)and1-b-D-glucopyranosyloxy-3-methoxy-5-hydroxybenzene(15)(Sakar et al.,1993)]compounds.Of the ten megastigmane glucosides,three were known com-pounds,namely spinoside A(8)(C¸alisßet al.,2002),staph-ylionoside D(9)(Yu et al.,2005),and corchoionoside C (10)(Yoshikawa et al.,1997).This paper deals with struc-tural elucidation of the seven new megastigmane glucosides.2.Results and discussionAir-dried leaves of E.meliaefolia were extracted with MeOH three times and the concentrated MeOH extract was partitioned with solvents of increasing polarity.The n-BuOH soluble fraction was separated by means of vari-ous chromatographic procedures including column chro-matography(CC)on a highly porous synthetic resin (Diaion HP-20),then normal silica gel and reversed-phase octadecyl silica gel(ODS)CC,droplet counter-current chromatography(DCCC),and high-performance liquid chromatography(HPLC)to afford15compounds(1–15). The details and yields are given under Section4.The struc-tures of the new megastigmane glucosides(17)were eluci-dated on the basis of spectroscopic evidence,that obtained with the modified Mosher’s method(Ohtani et al.,1991),and those of known compounds were identi-fied by comparison of spectroscopic data with those reported in the literature(Fig.1).The13C NMR spectro-scopic data of spinoside A(8)in CD3OD are included under Section4for the readers’convenience.Euodionoside A(1),½a 26D À40.5,was isolated as anamorphous powder and its elemental composition was determined to be C19H30O9by HR-ESI-TOF-MS.The IR spectrum indicated the presence of a sugar moiety (3395cmÀ1)and a ketonic functional group(1673cmÀ1), and the UV spectrum a conjugated system(235nm).The 1H and13C MR spectra showed the presence of six signalsassignable to b-glucopyranose,the remaining13carbon signals comprising four singlet methyls,two methylenes, one methine with a hydroxyl substituent,and four quater-nary carbons,and a disubstituted trans double bond,which must form the megastigmane skeleton.Judging from the chemical shifts,one(d H2.29)of the methyl groups must be adjacent to the carbonyl functional group,and one (d C200.1)and two(d C72.1and67.0)of the quaternary carbons must have a carbonyl group and oxygen substitu-ents,respectively.Whenfive degrees of unsaturation and a deficiency of one oxygen atom for three hydroxyl groups are considered,an epoxy ring must be required for one more cyclic system between C-5and C-6of megastigmane skeleton.In the H–H COSY spectrum,H2-2through H2-4 showed a significant correlation,and the correlations between H-7(d H7.08)and H-10(d H2.29),and the car-bonyl carbon(d C200.1)in the HMBC spectrum estab-lished a structure as1(Fig.1).The hydroxyl group at the C-3(d C72.1)position is the sole site to be glucosylated.To clarify the relative arrangement of the substituents,the phase-sensitive(PH)-NOESY spectrum was examined. Judging from the key correlations between H-3(d H3.98) and H3-11ax(d H1.25),H3-11and H-7(d H7.08),and H-4eq(d H2.38)and H3-13(d H1.17),H-3,the epoxy ring is in an anti orientation,in contrast to in the case of icariside B2,which was isolated from Epimedium grandiflorum var. thunbergianum(Miyase et al.,1987).On enzymatic hydro-lysis,an aglycone(1a)and D-glucose were obtained.From the b-D-glucosylation-induced shift-trend between the13C NMR spectra of1and1a,the absolute configuration of the3-position was determined to be R(Table1,a)(Kasai et al.,1977).This was further confirmed by the modified Mosher’s method(Fig.2)(Ohtani et al.,1991).Therefore, the structure of euodionoside A(1)was elucidated to be (3R,5R,6S,7E)-megastigman-7-en-5,6-epoxy-3-ol-9-one3-O-b-D-glucopyranoside,as shown in Fig.1.Euodionosides B(2),½a 25D24.8,and C(3),½a 26DÀ56.3, were isolated as amorphous powders and their elemental compositions were determined to be C19H32O8by HR-FAB-MS.The1H and13C NMR spectra indicated that euodionosides B(2)and C(3)were compounds analogous to euodionoside A(1),except for the presence of hydroxyl groups at their9-positions,instead of the carbonyl func-tional group found in1.The HMBC spectra showed a cor-relation between the anomeric proton(d H4.30)and C-3(d C 72.2)in the case of2,but between the former and(d H4.28) and C-9(d C74.8)in the case of3.The stereochemistry of the ring system of2was expected to be the same as that of1,since the13C NMR spectrum of2showed essentially the same chemical shifts as that of1.On enzymatic hydro-lysis of2,an aglycone(2a)(½aD+8.4)and D-glucose were obtained.The stereochemistry of the9-position of2a was found with the modified Mosher’s method to be S,and that of the ring portion was also confirmed by the PH-NOESY spectrum to comprise the3R,5R and6S configurations (Fig.2).The stereochemistry of the9-position of euodion-oside C(3)can be deduced with the b-D-glucosylation-induced shift-trend rule(Kasai et al.,1977)to be S,which is the same as that of2a,and that of the ring system was also expected to be the same as that of2.However,the ste-reochemistry of the ring system must be determined inde-pendently.Although the optical rotation value for theaglycone(3a)(½aD+5.2)of3was close to that of2a,the stereochemistry of2a was similarly established with the modified Mosher’s method(Fig.2).Therefore,the struc-tures of euodionosides B(2)and C(3)were elucidated to be(3R,5R,6S,7E,9S)-megastigman-7-en-5,6-epoxy-3,9-diol 3-and9-O-b-D-glucopyranosides,respectively.Euodionoside D(4),½aDÀ42.7,was isolated as an amorphous powder and its elemental composition was determined to be C19H34O9by HR-ESI-TOF-MS.On anal-yses of the NMR spectroscopic data,4was also expected to be a derivative of a megastigmane glucoside with four hydroxyl groups at the C-3,5,6,and9positions.The HMBC correlation between the anomeric proton(d H 4.38)and C-9(d C75.7)established that the sugar unitM.Yamamoto et al./Phytochemistry69(2008)1586–15961587was linked to the hydroxyl group at the9-position,and the coupling pattern of H-3(d H4.06,dddd,J=12,12,6,6Hz) indicated that the hydroxyl group at the3-position was in an equatorial orientation.From this evidence together with the results of comparison of the13C NMR spectroscopic data,the relative orientations of the substitutents of the ring system were found to be the same as those of (3S,5R,6R,7E,9S)-megastigman-7-ene-3,5,6,9-tetrol9-O-b-D-glucopyranoside isolated from Glochidion zeylanicum. According to the b-D-glucosylation-induced shift-trend rule (Kasai et al.,1977)the absolute configuration of the9-posi-tion was the same in the megastigmanes from E.meliaefo-lium and G.zeylanicum.However,although NMR data for the ring systems of these two compounds were essentially the same,the stereochemistries of the side chain and the ring system must be determined independently(Otsuka et al.,2003a).Thus,the modified Mosher’s method was applied to the aglycone(4a)(Fig.2),which was shown to have opposite absolute configurations to that from G.zey-lanicum.Therefore,the structure of euodionoside D(4) was elucidated to be(3R,5S,6S,7E,9S)-megastigman-7-ene-3,5,6,9-tetrol9-O-b-D-glucopyranoside,as shown in Fig.1.1588M.Yamamoto et al./Phytochemistry69(2008)1586–1596Table 113C NMR spectroscopic data for euodionosides A–G (1–7)(CD 3OD,100MHz)C 1234567135.935.835.940.740.937.537.3241.0(À3.3)a 41.444.446.446.348.648.7372.1(+8.2)a 72.263.865.365.1202.4202.2438.4(À1.6)a 38.640.245.738.7121.7123.3567.066.266.277.783.2172.3167.8672.172.272.179.179.248.447.97143.7124.3128.2135.5136.327.227.68134.6140.1136.9133.3132.737.4(À2.5)b 39.89200.168.774.875.775.777.7(+8.8)b 68.91027.324.022.522.622.521.9(À4.1)b 23.61125.025.125.327.527.627.727.71227.527.227.226.326.328.928.91321.021.822.227.720.365.270.9–O C H 348.710103.0102.9101.4100.7100.5104.0103.52075.175.175.175.175.075.475.13078.178.178.478.478.478.378.34071.871.871.871.971.771.771.75078.077.978.178.078.177.978.26063.062.963.063.062.862.962.9a D d 1–1a .bD d 6–7a .M.Yamamoto et al./Phytochemistry 69(2008)1586–15961589Euodionoside E(5),½aD À35.0,was isolated as anamorphous powder,and the one-and two-dimensional NMR spectroscopic data showed that it was a compound analogous to euodionoside D(4)with similar substituents on the six-membered ring.In the NMR spectra,a methoxyl signal was observed[d H3.21(3H,s)and d C48.7(q)],and the presence of an extra carbon atom was also supported by the HR-ESI-TOF-MS data(C20H36O9).From the HMBC spectroscopic correlation between d H3.21and d C 83.2(C-5),the position of the methoxyl group was assigned as C-5,and from the PH-NOESY spectrum,the relative orientations of the substituents were found to be the same as those of4.The absolute configurations of the ring sys-tem were also determined by the application of the modi-fied Mosher’s method to the aglycone(5a)(Fig.2). Therefore,the structure of euodionoside E(5)was eluci-dated to be(3R,5S,6S,7E,9S)-megastigman-7-ene-5-meth-oxy-3,6,9-triol9-O-b-D-glucopyranoside.A similar megastigmane derivative,which has a methoxyl group at the6-position,was isolated as staphylionoside J from Staphylea bumalda(Yu et al.,2005).Euodionosides F(6),½aD À0.7and G(7),½aD+5.9,were isolated as amorphous powders and their elemental compositions were determined to be C19H32O8by HR-ESI-TOF-MS.The IR(1649and1651cmÀ1,respectively) and UV(238and237nm,respectively)spectra showed the presence of a conjugated ketone,their NMR spectra being similar,with two singlet and one doublet methyls, and one primary and one secondary alcohol.Thus,one of the methyl groups was expected to be oxidized to a pri-mary alcohol,whose proton signals(d H4.16and4.32,and d H4.37and4.53,respectively)showed correlation to C-5 resonances(d C172.3and d C167.8)in the HMBC spectra. These facts were coincident with that C-13was oxidized to a primary alcohol.The HMBC spectra also indicated that6 and7were isomers of each other as to the positions of sugar linkages,the former had a sugar on the hydroxyl group at the9-position and the latter on that at the13-position,since correlation between anomeric protons and C-9(d C77.3)in6,and C-13(d C70.9)in7was observed, respectively.The absolute configuration of the9-position of6was determined to be R on comparison with a similar compound(Takeda et al.,1997)and that of the6-position was also assigned as R according to the circular dichroism (CD)spectral data(Otsuka et al.,2003b).The absolute configuration of the9-position was further supported by application of the b-D-glucosylation-induced shift-trend rule(Table1,b)(Otsuka et al.(1995)),Therefore,the struc-ture of euodionoside F(6)was elucidated to be(6R,9R)-megastigman-4-ene-9,13-diol9-O-b-D-glucopyranoside. Since euodionoside G(7)was an isomeric form of6,the absolute configuration of the9-position was expected to be the same as that of6.However,for the same reason as that for euodionoside D(4),the absolute configurations of the6-and9-positions were independently established by CD spectral analysis and the modified Mosher’s method (Fig.2),respectively.Although the exact D d S–d R values for the H2-7and eight protons could not be calculated, due to overlapping of their signals,they obviously had minus signs,whereas that for the H3-10protons had a sig-nificant plus value(+0.063)(Fig.2).As a result,the struc-ture of euodionoside G(7)was elucidated to be(6R,9R)-megastigman-4-ene-9,13-diol13-O-b-D-glucopyranoside.A compound closely related to6has been isolated from G.zeylanicum as glochidionionoside B,which has the9S configuration(Otsuka et al.,2003).3.Concluding remarksAlthough fruits of closely related species,E.rutaecarpa, are used for stomachic antipyretic and diuretic in tradi-tional Chinese medicine,there has been no report on the medicinal use of E.meloaefolia.Due to usage of leaves for this experiment,alkaloids are so far not isolated.Bio-logical evaluation and chemical investigation of fruits of E.meloaefolia will be subject of a future study.Megastigmane and its glycosides are currently an expanding class of compounds.Even with only13carbon atoms in the basic skeleton of megastigmane,several oxida-tion steps and glycosylation afforded many kinds of meg-astigmane derivatives and their glycosidic forms.In this experiment,seven new megastigmane glucosides were iso-lated from leaves of the title plant.Their stereostructures were established by the modified Mosher’s method.4.Experimental4.1.General experimental proceduresOptical rotations and CD spectra were measured on JASCO P-1030and JASCO J-720polarimeters,respec-tively.IR and UV spectra were measured on Horiba FT-710and JASCO V-520UV/Vis spectrophotometers, respectively.1H and13C NMR spectra were taken on a JEOL JNM a-400spectrometer at400MHz and 100MHz,respectively,with tetramethylsilane as an inter-nal standard.Negative-ion HR-MS was performed with a JEOL SX-102spectrometer in the FAB mode and posi-tive-ion HR-MS with an Applied Biosystem QSTAR XL system ESI(Nano Spray)-TOF-MS.A highly-porous synthetic resin(Diaion HP-20)was purchased from Mitsubishi Kagaku(Tokyo,Japan).Silica gel CC and reversed-phase[octadecyl silica gel(ODS)] open CC were performed on silica gel60(Merck,Darms-tadt,Germany)and Cosmosil75C18-OPN(Nacalai Tes-que,Kyoto,Japan)[U=50mm,L=25cm,linear gradient:MeOH–H2O(1:9,1L)?(1:1,1L),fractions of 10g being collected],respectively.Droplet counter-current chromatography(DCCC)(Tokyo Rikakikai,Tokyo, Japan)was equipped with500glass columns(U=2mm, L=40cm),and the lower and upper layers of a solvent mixture of CHCl3–MeOH–H2O–n-PrOH(9:12:8:2)were1590M.Yamamoto et al./Phytochemistry69(2008)1586–1596used for the stationary and mobile phases,respectively. Five-gram fractions were collected and numbered accord-ing to their order of elution with the mobile phase.HPLC was performed on an ODS column(Inertsil;GL Science, Tokyo,Japan;U=6mm,L=25cm),and the eluate was monitored with a UV detector at254nm and a refractive index monitor.Emulsin was purchased from Tokyo Chemical Indus-tries Co.Ltd.(Tokyo,Japan),and crude hesperidinase was a gift from Tokyo Tanabe Pharmaceutical Co.Ltd. (Tokyo,Japan).(R)-and(S)-a-methoxy-a-trifluoromethyl-phenylacetic acids(MTPA)were the products of Wako Pure Chemical Industry Co.Ltd.(Tokyo,Japan).4.2.Plant materialLeaves of E.meliaefolia Benth.(Rutaceae)were col-lected in Okinawa,Japan,in August2002,and a voucher specimen was deposited in the Herbarium of Pharmaceuti-cal Sciences,Graduate School of Biomedical Sciences, Hiroshima University(02-EM-Okinawa-0704).4.3.Extraction and isolationDried leaves of E.meliaefolia(15.7kg)were extracted three times with MeOH(45l)at25°C for one week and then concentrated to6l in vacuo.The extract was washed with n-hexane(6l,531g)and then the MeOH layer was concentrated to a gummy mass.The latter was suspended in water(6l)and then extracted with EtOAc(6l)to give 165g of an EtOAc-soluble fraction.The aqueous layer was extracted with1-BuOH(6l)to give a1-BuOH-soluble fraction(409g),and the remaining water-layer was concen-trated to furnish971g of a water-soluble fraction.A portion(154g)of the1-BuOH-soluble fraction was subjected to a Diaion HP-20column(U=50mm, L=50cm)using H2O–MeOH(4:1,8l),(2:3,8l),(3:2, 8l),and(1:4,8l),and MeOH(6l),1l fractions were being collected.The residue(27.6g in fractions6–12)of the20–40%MeOH eluent was subjected to silica gel(1.50kg) CC,with elution with CHCl3(6l)and CHCl3–MeOH [(99:1,6l),(97:3,6l),(19:1,6l),(37:3,6l),(9:1,6l),(7:1, 6l),(17:3,6l),(33:7,6l),(4:1,6l),(31:9,6l),(3:1,6l), and(7:3,6l)],1l fractions being bined frac-tions31–39(1.20g)were separated by RPCC.The residue (383mg)of fractions62–78was subjected to DCCC to give 25.4mg of11in fractions35–40,and the residue(44.5mg) in fractions44–52was further purified by prep.HPLC with MeOH–H2O(3:7)to give7.6mg of13at R t10min.The residue(112mg)in fraction94–114obtained on RPCC was subjected to DCCC to give3.9mg of1in fractions 63–72,and the residue(10.6mg in fractions21–27)was fur-ther purified by prep.HPLC with MeOH–H2O(3:7)to give 3.9mg of4at R t13min.The residue(85mg)in fractions 115–132obtained on RPCC was also subjected to DCCC to give two fractions,10.2mg in fractions90–105and 37.3mg in fractions109–136.Prep.HPLC of the former fraction with CH3CN–H2O(3:17)gave1.4mg of6and 2.8mg of7at R t s16min and18min,respectively.The lat-ter was found to be a pure compound,14.The residue(1.50g out of3.03g)in fractions40–45 obtained on15–17.5%MeOH eluate on silica gel CC was subjected to RPCC to givefive fractions,172mg in frac-tions37–47,87.0mg in fractions62–65,150mg in fractions 81–89,116mg in fractions98–104,and101mg in fractions 112–123.Thefirst fraction was purified by DCCC to give 144mg of15in fractions21–29.The second fraction was purified by DCCC(40.1mg in fractions47–57)and then prep.HPLC with MeOH–H2O(1:4)to afford5.4mg of8 at R t30min.The third fraction was subjected to prep. HPLC with MeOH–H2O(1:3)to give two partially purified fractions at R t s34min(36.0mg)and42min(59.4mg).The former was repeatedly purified by prep.HPLC with CN3CN–H2O(1:9)to give15.3mg of9and5.2mg of10 at R t s56min and68min,respectively.The latter gave 21.0mg of2on prep.HPLC with CH3CN–H2O(13:87) at R t44min.The residue of the fourth fraction was found to be a pure compound,3.Thefifth fraction was subjected to prep.HPLC with MeOH–H2O(19:31)to give8.5mg of 6at R t18min.The residue(1.61g)in fractions46–51obtained from the17.5–20%MeOH eluate on silica gel CC was subjected to RPCC to give two fractions,130mg in fractions40–46 and20.0mg in fractions146–153.The former was further purified by DCCC to give29.6mg of15in fractions53–58and the latter was a pure compound,5.4.4.Characterization data4.4.1.Euodionoside A(1)Amorphous powder;½a 26DÀ40.5(c=0.25,MeOH);IR m max(film)cmÀ1:3395,2926,1673,1455,1367,1076;UV k max(MeOH)nm(log e):235(3.75);1H NMR(CD3OD, 400MHz)d:7.08(1H,d,J=16Hz,H-7),6.20(1H,d, J=16Hz,H-8),4.31(1H,d,J=8Hz,H-10),3.98(1H, dddd,J=11,10,7,7Hz,H-3), 3.87(1H,dd,J=12, 2Hz,H-60a), 3.65(1H,dd,J=12,6Hz,H-60b), 3.42–3.25(3H,m,H-30,40and50),3.12(1H,dd,J=9,8Hz, H-20), 2.38(1H,dd,J=15,7Hz,H-4pseudo-eq), 2.29 (3H,s,H3-10),1.98(1H,dd,J=15,10Hz,H-4pseudo-ax), 1.50(1H,dd,J=11,7Hz,H-2eq), 1.49(1H,dd, J=11,11Hz,H-2ax),1.25(3H,s,H3-11),1.17(3H,s, H3-13),0.97(3H,s,H3-12);13C NMR(CD3OD, 100MHz):Table1;HR-ESI-TOF-MS(positive-ion mode) m/z:409.1856[M+Na]+(calcd for C19H30O8Na,409.1832).4.4.2.Euodionoside B(2)Amorphous powder;½a 25D–24.8(c=0.79,MeOH);IR m max(film)cmÀ1:3367,2965,1653,1453,1379,1075;1H NMR(CD3OD,400MHz)d:5.84(1H,dd,J=16,1Hz, H-7), 5.67(1H,dd,J=16,6Hz,H-8), 4.30(1H,d, J=8Hz,H-10),4.29(1H,qdd,J=6,6,1Hz,H-9),3.94 (1H,dddd,J=11,10,7,6HZ,H-3), 3.86(1H,dd, J=12,2Hz,H-6b),3.65(1H,dd,J=12,6Hz,H-60b),M.Yamamoto et al./Phytochemistry69(2008)1586–159615913.36–3.25(3H,m,H-30,40and50),3.12(1H,dd,J=9, 8Hz,H-20),2.32(1H,ddd,J=15,7,1Hz,H-4pseudo-eq), 1.89(1H,dd,J=15,10Hz,H-4pseudo-ax), 1.49 (2H,m,H2-2),1.23(3H,d,J=6Hz,H3-10),1.20(3H,s, H3-13), 1.16(3H,s,H3-11),0.97(3H,s,H3-12);13C NMR(CD3OD,100MHz):Table1;HR-FAB-MS(nega-tive-ion mode)m/z:387.2028[MÀH]À(calcd for C19H31O8,387.2019).4.4.3.Euodionoside C(3)Amorphous powder;½a 24D À56.3(c=7.20,MeOH);IRm max(film)cmÀ1:3395,2965,1649,1452,1368,1075;1H NMR(CD3OD,400MHz)d:6.01(1H,dd,J=16,1Hz, H-7),5.57(1H,ddd,J=16,7,1Hz,H-8),4.52(1H,br dq,J=7,6Hz,H-9),4.28(1H,d,J=8Hz,H-10),3.86 (1H,dd,J=12,2Hz,H-60a), 3.81(1H,m,H-3), 3.64 (1H,dd,J=12,6Hz,H-60b),3.32–3.14(4H,m,H-20,30, 40and50), 2.21(1H,dd,J=15,7Hz,H-4pseudo-eq), 1.71(1H,dd,J=15,10Hz,H-4pseudo-ax),1.46(1H,dd, J=12,12Hz,H-2ax), 1.28(3H,d,J=6Hz,H3-10), 1.27(1H,m,H-2eq),1.23(3H,s,H3-13),1.17(3H,s,H3-11),0.96(3H,s,H3-12);13C NMR(CD3OD,100MHz): Table1;HR-FAB-MS(negative-ion mode)m/z:387.2021 [MÀH]À(calcd for C19H31O8,387.2018).4.4.4.Euodionoside D(4)Amorphous powder;½a 24D À42.7(c=0.28,MeOH);IRm max(film)cmÀ1:3395,2927,1649,1372,1072,1027;1H NMR(CD3OD,400MHz)d:6.16(1H,d,J=16Hz,H-7),5.63(1H,dd,J=16,8Hz,H-8),4.52(1H,dq,J=8, 7Hz,H-9),4.38(1H,d,J=8Hz,H-10),4.06(1H,dddd, J=12,12,6,6Hz,H-3),3.87(1H,dd,J=12,2Hz,H-60a),3.65(1H,dd,J=12,6Hz,H-60b),3.35–3.18(4H, m,H-20,30,40and50),1.76(2H,m,H2-4),1.65(1H,dd, J=12,12Hz,H-2ax),1.45(1H,ddd,J=12,4,2Hz,H-2eq),1.31(3H,d,J=7Hz,H3-10),1.173(3H,s,H3-12), 1.165(3H,s,H3-13),0.85(3H,s,H3-11);13C NMR (CD3OD,100MHz):Table1;HR-ESI-TOF-MS(posi-tive-ion mode)m/z:429.2098[M+Na]+(calcd for C19H34O9Na,429.2095).4.4.5.Euodionoside E(5)Amorphous powder;½a 24D À35.0(c=1.13,MeOH);IRm max(film)cmÀ1:3395,2931,1635,1370,1077,1035;1H NMR(CD3OD,400MHz)d:6.13(1H,d,J=16Hz,H-7),5.59(1H,dd,J=16,9Hz,H-8),4.49(1H,dq,J=9, 6Hz,H-9),4.37(1H,d,J=8Hz,H-10),3.85(1H,dd, J=12,2Hz,H-60a),3.83(1H,dddd,J=12,12,4,4Hz, H-3),3.67(1H,dd,J=12,6Hz,H-60b),3.33–3.18(4H, m,H-20,30,40and50),3.21(3H,s,–OCH3),2.10(1H, ddd,J=12,4,2Hz,H-4eq),1.65(1H,dd,J=12,12Hz, H-2ax),1.57(1H,dd,J=14,12Hz,H-4ax),1.45(1H, ddd,J=12,4,2Hz,H-2eq),1.30(3H,d,J=6Hz,H3-10),1.11(3H,s,H3-13),1.10(3H,s,H3-12),0.85(3H,s, H3-11);13C NMR(CD3OD,100MHz):Table1;HR-ESI-TOF-MS(positive-ion mode)m/z:443.2244 [M+Na]+(calcd for C20H36O9Na:443.2251).4.4.6.Euodionoside F(6)Amorphous powder;½a 24DÀ0.70(c=0.55,MeOH);IR m max(film)cmÀ1:3396,2930,1649,1076,1037;UV k max (MeOH)nm(log e):238(3.88);1H NMR(CD3OD, 400MHz)d:6.60(1H,br s,H-4),4.33(1H,d,J=8Hz, H-10),4.32(1H,dd,J=18,1Hz,H-13a),4.16(1H,dd, J=18,1Hz,H-13b),3.84(1H,m,H-60a),3.82(1H,m, H-9),3.67(1H,dd,J=11,6Hz,H-60b),3.36–3.26(3H, m,H-30,40and50),3.15(1H,dd,J=9,8Hz,H-20),2.58 (1H,d,J=17Hz,H-2a),2.03(1H,d,J=17Hz,H-2b), 1.92(1H,m,H-6),1.80-1.52(4H,m,H2-7and8),1.24 (3H,d,J=6Hz,H3-10),1.11(3H,s,H3-12),1.02(3H,s, H3-11);13C NMR(CD3OD,100MHz):Table1;CD nm (D e):219(+2.24),278(À0.11),335(+0.51) (c=3.94Â10À5M,MeOH);HR-ESI-TOF-MS(positive-ion mode)m/z:411.1983[M+Na]+(calcd for C19H32O8Na, 411.1989).4.4.7.Euodionoside G(7)Amorphous powder;½a 27D+5.9(c=0.19,MeOH);IR m max(film)cmÀ1:3367,2927,1651,1077,1040;UV k max (MeOH)nm(log e):237(3.57);1H NMR(CD3OD, 400MHz)d:6.17(1H,br s,H-4),4.53(2H,dd,J=18, 2Hz,H-13a), 4.37(1H,dd,J=18,2Hz,H-13b), 4.33 (1H,d,J=8Hz,H-10),3.88(1H,dd,J=12,2Hz,H-60a), 3.68(1H,m,H-9), 3.62(1H,dd,J=12,6Hz,H-60b), 3.36–3.15(4H,m,H-20,30,40and50), 2.54(1H,d, J=18Hz,H-2a),2.03(1H,t,J=6Hz,H-6),2.03(1H,d, J=18Hz,H-2b),1.80–1.52(4H,m,H2-7and8),1.60(3H, d,J=6Hz,H3-10),1.11(3H,s,H3-11),1.03(3H,s,H3-12);13C NMR(CD3OD,100MHz):Table1;CD nm(D e): 235(+1.78),276(À0.21),337(+0.29)(c=3.22Â10À5M, MeOH);HR-ESI-TOF-MS(positive-ion mode)m/z: 411.1995[M+Na]+(calcd for C19H32O8Na,411.1989).4.4.8.Enzymatic hydrolysis of euodionosides A(1)–E(5) and G(7)Euodionoside A(1)(3.8mg)in2ml of H2O was hydro-lyzed with emulsin(6.0mg)and crude hesperidinase (6.0mg)for15h at37°C.The reaction mixture was evap-orated to dryness,and then the methanolic solution was absorbed on silica gel and subjected to silica gel CC (20g,U=18mm,L=18cm)with CHCl3(100ml)and CHCl3–MeOH(19:1,100ml,9:1,100ml,17:3,100ml and7:3,300ml),12ml fractions being collected.An agly-cone(1a)(1.8mg,82%)and D-glucose(1.3mg,77%)were recovered in fractions15–25and37–47,respectively.Agly-cone(1a):An amorphous powder,½a 25D+27.9(c=0.12, MeOH);1H NMR(CD3OD,400MHz)d:7.08(1H,d, J=16Hz,H-7),6.20(1H,d,J=16Hz,H-8),3.85(1H, dddd,J=12,10,7,4Hz,H-3),2.28(3H,s,H3-10),2.24 (1H,ddd,J=15,7,2Hz,H-4pseudo-eq),1.73(1H,dd, J=15,10Hz,H-4pseudo-ax), 1.48(1H,dd,J=12, 12Hz,H-2ax), 1.33(1H,ddd,J=12,4,2Hz,H-2eq), 1.24(3H,s,H3-11)1.17(3H,s,H3-13),0.96(3H,s,H3-12);13C NMR(CD3OD,100MHz)d:143.8(C-7),134.6 (C-8),63.9(C-3),44.3(C-2),40.0(C-4),36.0(C-1),27.61592M.Yamamoto et al./Phytochemistry69(2008)1586–1596(C-12),27.3(C-10),25.2(C-11),21.4(C-13),singlet car-bons(C-5,6and9)were not observed;HR-ESI-TOF-MS (positive-ion mode)m/z:247.1307[M+Na]+(calcd forC13H20O3Na,247.1304).D-Glucose:½a 25D +32.2°(c=0.087,H2O,24h after being dissolved in the solvent).From euodionoside B(2)(12mg),6.8mg(91%)of an aglycone(2a)and4.9mg(83%)of D-glucose were obtained:Aglycone(2a):an amorphous powder;½a 25D +8.4(c=0.31,MeOH);1H NMR(CD3OD,400MHz)d:5.85(1H,dd, J=16,1Hz,H-7),5.67(1H,dd,J=16,6Hz,H-8),4.30 (1H,qd,J=6,1Hz,H-9),3.81(1H,dddd,J=12,10,7, 4Hz,H-3),2.20(1H,ddd,J=15,7,2Hz,H-4pseudo-ax),1.71(1H,dd,J=15,10Hz,H-4pseuo-eq),1.46(1H, dd,J=12,12Hz,H-2ax),1.28(1H,ddd,J=12,4,2Hz, H-2eq),1.23(3H,d,J=6Hz,H3-10),1.20(3H,s,H3-13),1.15(3H,s,H3-11),0.95(3H,s,H3-12);13C NMR (CD3OD,100MHz)d:140.1(C-8),124.4(C-7),72.0(C-6),68.7(C-9),66.1(C-5),63.8(C-3),44.5(C-2),40.2(C-4),36.0(C-1),27.2(C-12),25.3(C-11),23.9(C-10),21.8 (C-13);HR-ESI-TOFMS(positive-ion mode)m/z: 249.1466[M+Na]+(calcd for C13H22O5Na,249.1461).D-Glucose,½a 24D +41.3°(c=0.33,H2O,24h after being dis-solved in the solvent).From euodionoside B(3)(17mg),5.2mg(49%)of an aglycone(3a)and5.8mg(69%)of D-glucose were obtained.Aglycone(3a):An amorphous powder;½a 24D +5.2(c=0.41,MeOH);1H NMR(CD3OD,400MHz)d:5.85(1H,dd,J=16,1Hz,H-7),5.67(1H,dd,J=16,6Hz,H-8),4.30 (1H,qd,J=6,1Hz,H-9),3.81(1H,dddd,J=12,10,7, 4Hz,H-3),2.20(1H,ddd,J=15,7,2Hz,H-4pseudo-eq), 1.71(1H,dd,J=15,10Hz,H-4pseudo-ax), 1.46 (1H,dd,J=12,12Hz,H-2ax),1.28(1H,ddd,J=12,4, 2Hz,H-2eq),1.23(3H,d,J=6Hz,H3-10),1.20(3H,s, H3-13), 1.15(3H,s,H3-11),0.95(3H,s,H3-12);13C NMR(CD3OD,100MHz)d:140.1(C-8),124.4(C-7), 72.0(C-6),68.7(C-9),66.1(C-5),63.8(C-3),44.5(C-2), 40.3(C-4),36.0(C-1),27.2(C-12),25.3(C-11),23.9(C-10),21.8(C-13);HR-ESI-TOFMS(positive-ion mode)m/ z:249.1466[M+Na]+(calcd for C13H22O5Na,249.1464). D-Glucose,½a 27D+51.6(c=0.38,H2O,24h after being dis-solved in the solvent).From euodionoside B(4)(3.8mg),1.8mg(79%)of an aglycone(4a)and 1.1mg(65%)of D-glucose wereobtained.Aglycone(4a):An amorphous powder;½a 30D +18.6(c=0.19,MeOH);1H NMR(CD3OD,400MHz) d: 6.06(1H,d,J=16Hz,H-7), 5.63(1H,dd,J=16, 6Hz,H-8), 4.34(1H,qd,J=6,1Hz,H-9), 4.05(3H, m,H-3), 1.78(1H,ddd,J=13,5,2Hz,H-4eq), 1.73 (1H,dd,J=13,12Hz,H-4ax), 1.65(1H,dd,J=12, 12Hz,H-2ax), 1.44(1H,ddd,J=12,4,2Hz,H-2eq), 1.27(3H,d,J=6Hz,H3-10),1.20(3H,s,H3-12),1.14 (3H,s,H3-13),0.84(3H,s,H3-11);13C NMR(CD3OD, 100MHz)d:136.2(C-7),131.3(C-8),83.6(C-6),77.8 (C-5),69.6(C-9),65.3(C-3),46.5(C-2),45.8(C-4), 40.8(C-1),27.5(C-11),27.1(C-13),26.3(C-12),24.3 (C-10);HR-ESI-TOFMS(positive-ion mode)m/z: 267.1554[M+Na]+(calcd for C13H24O4Na,247.1566).D-Glucose,½a 27D+25.0(c=0.073,H2O,24h after being dissolved in the solvent).From euodionoside B(5)(10.4mg),5.3mg(83%)of an aglycone(5a)and3.5mg(78%)of D-glucose were obtainedAglycone(5a):An amorphous powder;½a 25D+25.5°(c=0.35,MeOH);1H NMR(CD3OD,400MHz)d:5.95 (1H,d,J=16Hz,H-7),5.65(1H,dd,J=16,9Hz,H-8), 4.21(1H,m,H-9),3.73(1H,m,H-3),3.09(3H,s,-OMe), 1.99(1H,ddd,J=14,4,2Hz,H-4eq), 1.55(1H,dd, J=12,12Hz,H-2ax),1.45(1H,dd,J=14,12Hz,H-4ax),1.35(1H,ddd,J=12,4,2Hz,H-2eq),1.15(3H,d, J=6Hz,H3-10),1.02(3H,s,H3-12),0.97(3H,s,H3-13),0.74(3H,s,H3-11);13C NMR(CD3OD,100MHz): d135.6(C-7),131.5(C-8),83.3(C-5),79.0(C-6),69.7(C-9),65.2(C-3),48.4(–OCH3),46.5(C-2),40.9(C-1),38.8 (C-4),27.7(C-11),26.4(C-12),24.2(C-10),19.7(C-13); HR-ESI-TOFMS(positive-ion mode)m/z:281.1729 [M+Na]+(calcd for C14H26O4Na,281.1723).D-Glucose,½a 25D+49.7(c=0.23,H2O,24h after being dissolved in the solvent).From euodionoside G(7)(2.5mg),1.2mg(82%)of an aglycone(7a)and1.0mg(86%)of D-glucose were obtained.Aglycone(7a):An amorphous powder;½a 25D+61.7°(c=0.08,MeOH);1H NMR(CD3OD,400MHz)d:6.06 (1H,br s,H-4),4.30(1H,dd,J=18,2Hz,H-13),4.15 (1H,dd,J=18,2Hz,H-13),3.66(1H,qd,J=6,6Hz, H-9), 2.54(1H,dd,J=18,1Hz,H-2), 2.03(1H,ddd, J=18,1,1Hz,H-2), 1.94(1H,dd,J=5,5Hz,H-6), 1.821.49(4H,m,H2-7,8),1.16(3H,d,J=6Hz,H3-10), 1.11(3H,s,H3-11),1.02(3H,s,H3-12);13C NMR (CD3OD,100MHz)d:172.3(C-5),121.7(C-4),68.9 (C-9),65.1(C-13),48.7(C-2),48.4(C-6),39.9(C-8),37.4 (C-1),28.9(C-12),28.0(C-7),27.6(C-11),23.6(C-10); HR-ESI-TOF-MS(positive-ion mode)m/z:249.1465 [M+Na]+(calcd for C13H22O2Na,249.1461).D-Glucose,½a 27D+20.4(c=0.07,H2O,24h after being dissolved in the solvent).4.4.9.Preparation of(R)-and(S)-MPTA estersA solution of1a(0.9mg)in1ml of dehydrated CH2Cl2was reacted with(R)-MTPA(27mg)in the pres-ence of1-ethyl-3-(3-dimethylaminopropyl)cardodiimide hydrochloride(EDC)(16mg)and N,N-dimethyl-4-ami-nopyridine(4-DMAP)(11mg),and then the mixture was occasionally stirred at25°C for30min.After the addition of1ml of CH2Cl2,the solution was washed with H2O(1ml),5%HCl(1ml),NaHCO3–saturated H2O,and then brine(1ml),successively.The organic layer was dried over Na2SO4and then evaporated under reduced pressure.The residue was purified by preparative TLC[silica gel(0.25mm thickness),being applied for 18cm,developed with CHCl3–(CH3)2CO(19:1)for 9cm,and then eluted with CHCl3–MeOH(9:1)]to fur-nish an ester,1b(1.2mg,68%).Through a similar proce-dure,1c(1.3mg,73%)was prepared from1a(0.9mg) using(S)-MTPA(27mg),EDC(17mg),and4-DMAP (12mg).M.Yamamoto et al./Phytochemistry69(2008)1586–15961593。
核磁共振图谱解析解析NMR
同核J-偶合(Homonuclear J-Coupling)
多重峰出现的规则: 1. 某一原子核与N个相邻的核相互偶合将给出(n+1)重峰. 2. 等价组合具有相同的共振频率.其强度与等价组合数有关. 3. 磁等价的核之间偶合作用不出现在谱图中. 4. 偶合具有相加性. 例如: observed spin coupled spin intensity
JCH JCH
H C
p-pulse on H
H C
这相当于使用一系列1800脉冲快速照射氢核。 pH pH
C-H
+J/2
C-H
-J/2
C-H
+J/2
pH
C-H
-J/2
pH
C-H
+J/2
pH
C-H
-J/2
Fig. 4-2.5 The proton-decoupled 13C spectrum of 1-propanol
H-12C H-13C H-13C x100
105 Hz
proton-coupled spectra (nondecoupled spectra)
Quartet, J=127 Hz
Proton-coupled spectra for large molecules are often difficult to interpret. The multiplets from different C commonly overlap because the 13C-H coupling constants are frequently larger than the chemical differences of the C in the spectrum. 原子核间的偶合导致谱图 的复杂化(―精细裂分”)， 灵敏度下降。 Fig. 4-2.4 Ethyl phenylacetate. (a) The proton-coupled 13C spectrum. (b) The proton-decoupled 13C spectrum
光谱法研究药物小分子与蛋白质大分子的相互作用的英文
Spectroscopic Study of the Interaction between Small Molecules and Large Proteins1. IntroductionThe study of drug-protein interactions is of great importance in drug discovery and development. Understanding how small molecules interact with proteins at the molecular level is crucial for the design of new and more effective drugs. Spectroscopic techniques have proven to be valuable tools in the investigation of these interactions, providing det本人led information about the binding affinity, mode of binding, and structural changes that occur upon binding.2. Spectroscopic Techniques2.1. Fluorescence SpectroscopyFluorescence spectroscopy is widely used in the study of drug-protein interactions due to its high sensitivity and selectivity. By monitoring the changes in the fluorescence emission of either the drug or the protein upon binding, valuable information about the binding affinity and the binding site can be obt本人ned. Additionally, fluorescence quenching studies can provide insights into the proximity and accessibility of specific amino acid residues in the protein's binding site.2.2. UV-Visible SpectroscopyUV-Visible spectroscopy is another powerful tool for the investigation of drug-protein interactions. This technique can be used to monitor changes in the absorption spectra of either the drug or the protein upon binding, providing information about the binding affinity and the stoichiometry of the interaction. Moreover, UV-Visible spectroscopy can be used to study the conformational changes that occur in the protein upon binding to the drug.2.3. Circular Dichroism SpectroscopyCircular dichroism spectroscopy is widely used to investigate the secondary structure of proteins and to monitor conformational changes upon ligand binding. By analyzing the changes in the CD spectra of the protein in the presence of the drug, valuable information about the structural changes induced by the binding can be obt本人ned.2.4. Nuclear Magnetic Resonance SpectroscopyNMR spectroscopy is a powerful technique for the investigation of drug-protein interactions at the atomic level. By analyzing the chemical shifts and the NOE signals of the protein in thepresence of the drug, det本人led information about the binding site and the mode of binding can be obt本人ned. Additionally, NMR can provide insights into the dynamics of the protein upon binding to the drug.3. Applications3.1. Drug DiscoverySpectroscopic studies of drug-protein interactions play a crucial role in drug discovery, providing valuable information about the binding affinity, selectivity, and mode of action of potential drug candidates. By understanding how small molecules interact with their target proteins, researchers can design more potent and specific drugs with fewer side effects.3.2. Protein EngineeringSpectroscopic techniques can also be used to study the effects of mutations and modifications on the binding affinity and specificity of proteins. By analyzing the binding of small molecules to wild-type and mutant proteins, valuable insights into the structure-function relationship of proteins can be obt本人ned.3.3. Biophysical StudiesSpectroscopic studies of drug-protein interactions are also valuable for the characterization of protein-ligandplexes, providing insights into the thermodynamics and kinetics of the binding process. Additionally, these studies can be used to investigate the effects of environmental factors, such as pH, temperature, and ionic strength, on the stability and binding affinity of theplexes.4. Challenges and Future DirectionsWhile spectroscopic techniques have greatly contributed to our understanding of drug-protein interactions, there are still challenges that need to be addressed. For instance, the study of membrane proteins and protein-protein interactions using spectroscopic techniques rem本人ns challenging due to theplexity and heterogeneity of these systems. Additionally, the development of new spectroscopic methods and the integration of spectroscopy with other biophysical andputational approaches will further advance our understanding of drug-protein interactions.In conclusion, spectroscopic studies of drug-protein interactions have greatly contributed to our understanding of how small molecules interact with proteins at the molecular level. Byproviding det本人led information about the binding affinity, mode of binding, and structural changes that occur upon binding, spectroscopic techniques have be valuable tools in drug discovery, protein engineering, and biophysical studies. As technology continues to advance, spectroscopy will play an increasingly important role in the study of drug-protein interactions, leading to the development of more effective and targeted therapeutics.。
化学专业英语5-experiment
dropwise to a solution of 1,3-propanediol (29.9 g,
0.393 mol) and ethyl chloroformate (85.4 g, 0.786
mol) in 2L of THF at 0 oC over a period of 30 min.
Generally, an experimental section mainly consists
three parts, that is:
Chemicals (Materials), Synthesis (preparation),
and Characterization
Experimental
PVA is poly (vinyl alcohol) 聚乙烯醇
一、原材料的来源及准备(Chemicals or Materials)
Hale Waihona Puke All materials, such as potassium hexacyano-cobaltate (III) (K3Co(CN)6), zinc fluoride (ZnF2), zinc chloride (ZnCl2), zinc bromide (ZnBr2), zinc iodide (ZnI2) and tertiary butyl alcohol (tBuOH) were purchased from Aldrich and used without further purification.
一、原材料的来源及准备(Chemicals or Materials) 2. 直接购买的普通溶剂或辅助药品只需简单提及或略 过
核磁共振波谱课程教学探索
山东化工SHANDONGCHEMICALINDUSTRY-158-2020年第49卷核磁共振波谱课程教学探索李晓虹(苏州大学材料与化学化工学部，江苏苏州215123)摘要:核磁共振波谱作为鉴定化合物结构、组分含量、动力学参数等信息的重要手段,在化学、医药、材料等领域科研生产中起着关键作用。
其课程教学长期以来受到理论内容难、仪器开放难等因素困扰’结合苏州大学核磁共振波谱课程的双语教学实践提出了相应的对策与改进举措，探讨通过更新改进教学方法和内容，突破传统教学模式，使学生从理论联系实践,从“会用”到“用好”核磁技术’关键词:核磁共振波谱;远程虚拟终端%网络课堂中图分类号：G642O文献标识码：B文章编号：1008-021X(2020)23-0158-02Exploration of Teaching in Nuclear Magnetic Resonance Spectroscopy CourseLi Xiaohong(Colleae of Chemist—,Chemicai Enginee/ng and Materials Science of Soochow University,Suzhou215123,China) Abstract:Nuclear magnetic resonance spectroscopy(NMR),as an Onportant method of studying compound structures, component contents and kinetic parameters,plays a key rolo in the fields of chemist—,pharmaceutical indust—and materials science.For a long time,its course teaching has been troubled by the dOficulta of theo—tical content and the lack of instmmentai peacicce.Based on ihebcocnguaoieachcngpeacicceooNMR couesecn Soochow Unceeesciy,ihcspapeedcscu s eshow iobeeak iheough iheieadciconaoieachcngmodebycmpeoecngiheieachcngmeihodsand conienis,soihaisiudeniscan combcneiheoeywcih peacicceand makegood useooNMRiechnooogy.Key wordt:NMR%VNC%online coa s es核磁共振波谱作为鉴定化合物结构的重要手段，对样品无损，分辨率高,较灵敏，可获得准确的定性定量信息。
CIESCJ2015-66(9)-P3719-3725-ZhangR-Lzwc-利用_13_CNMR技术探究叔胺溶液中HCO_3_的生成
2015年9月 CIESC JournalSeptember 2015第66卷 第9期 化 工 学 报 V ol.66 No.9利用13C NMR 技术探究叔胺溶液中3HCO −的生成张瑞，李末霞，罗潇，梁志武（CO 2捕获与封存国际合作中心（iCCS ），化石能源低碳化高效利用湖南省重点实验室，湖南大学化学化工学院，湖南 长沙410082）摘要：利用13C NMR 技术对CO 2捕获叔胺溶剂进行了碳元素的定量研究，主要考察了对胺溶剂解吸热影响较大的3HCO −的生成规律。
重点对叔胺分子结构中羟基官能团（OH ）、羟烷基数目、烷基支链及氮原子（N ）所连接烷基链大小对胺溶剂生成3HCO −的影响。
在20℃条件下分别对1 mol ·L −1具有不同CO 2负载的1-二甲基氨基-2-丙醇（1DMA2P ）、N -甲基二乙醇胺（MDEA ）、3-二甲氨基-1-丙醇（3DMA1P ）、二乙氨基-2-丙醇（1DEA2P ）、N ,N -二乙基乙醇胺（DEEA ）、N ,N -二甲基乙醇胺（DMMEA ）及三乙醇胺（TEA ）溶液进行了13C NMR 测试研究。
实验结果显示：在相同浓度的叔胺水溶液中，同一CO 2负载下的叔胺-CO 2-水体系中3HCO −的含量顺序为：DMMEA ＞ MDEA ＞3DMA1P ＞1DMA2P ＞TEA ＞DEEA ＞1DEA2P 。
通过对各叔胺分子结构中N 原子的电子云密度大小及空间位阻效应分析，得出如下结论：3DMA1P 分子中OH 官能团离N 原子的距离大于其在DMMEA 分子中的距离，导致其生成了较少的3HCO −；DMMEA 分子中N 原子上连接的烷基链小于DEEA 分子中N 原子上的烷基链，导致DMMEA 溶液中生成了更多3HCO −；MDEA 分子中羟烷基数目少于TEA 分子中的羟烷基数目，且MDEA 比TEA 多了一个甲基，导致MDEA 溶液中含有更多的3HCO −；3DMA1P 相比1DMA2P 、DEEA 相比1DEA2P 分子中都少了一个甲基支链，导致3DMA1P 溶液相比1DMA2P 溶液、DEEA 溶液相比1DEA2P 溶液生成了更多的3HCO −。
黄花远志的新齐墩果烷型三萜皂甙_英文_
黄花远志的新齐墩果烷型三萜皂甙吴志军 欧阳明安 杨崇仁#(中国科学院昆明植物研究所,昆明 650204)摘要 从云南产远志科药用植物黄花远志(Polygala arillata Buch .-Ham .ex D .Don )茎皮的乙醇提取物中分离得到4个新的齐墩果烷型三萜皂甙,命名为黄花远志皂甙(arillatanoside )A ～D 。
同时还分离得到1个已知的三萜皂甙远志甙(polygalasaponin )XXXV 。
它们的结构通过波谱方法推定。
关键词 远志科,黄花远志,三萜皂甙,黄花远志皂甙A ～D 分类号 Q 946New Oleanane Triterpenoid Saponins from Polygala arillata ＊WU Zhi -Jun ,OUYANG Ming -An ,YANG Chong -Ren #(Kunming Institute of Bot any ,The C hine se Academy of Sci ence s ,Kunming 650204)A bstract Four new oleanane triterpenoidal saponins ,arillatanoside A ～D ,together with a known saponin polygalasaponin XXXV were isolated from the stem bark of Polygala arillata .The structures of new saponins were established to be 28-O -α-L -arabinopyranosyl -(1※3)-β-D -xylopyranosyl -(1※4)-α-L -rhamnopyranosyl -(1※2)-β-D -fucopyranosyl presenegenin -3-O -β-D -glucopyranoside ,28-O -β-D -galactop yranos yl -(1※4)-[α-L -arabinopyranosyl -(1※3)]-β-D -xylopyranosyl -(1※4)-α-L -rhamnopyranosyl -(1※2)-[4-O -acetyl ]-β-D -fucopyranosyl presenegenin -3-O -β-D -glucopyranoside ,28-O -β-D -galactopyranosyl -(1※4)-[α-L -arabinopyranosyl -(1※3)]-β-D -xylop y ranos y l -(1※4)-α-L -rhamnopyranosyl -(1※2)-β-D -fucopyranosyl presenegenin 3-O -β-D -glucopyranoside and 28-O -β-D -galactopyranosyl -(1※4)-[α-L -arabinop yranos yl -(1※3)]-β-D -xylopyranosyl -(1※4)-[β-D -apiofurarnosyl -(1※3)]-α-L -rhamnop yranos yl -(1※2)-β-D -fucop yranos yl presenegenin -3-O -β-D -glucopyranoside ,re -spectel y ,by FAB -MS and NMR spectroscopy .Key words Pol ygalaceae ,Polygala arillata ,Triterpenoidal saponins ,Arillatanoside A ～DPolygala arillata Buch .-Ham .ex D .Don is a moderate size tree in the family Polygalaceae ,distributed in southern China .It as a folk herb is used for treating coughs ,expectorants ,stomach trou -ble and rheumatism (Jiangsu College of New Medicine ,1979).Chemical studies on this plant have in -云南植物研究 1999,21(3):357～363Acta Botanica Yunnanica 云南省科委应用基础基金资助项目 This work was s upported by grand of Scientific Foundation of Y unnan (China )#通信联系人 Author to whom corres pondence shoul d be addressed .1998-11-04收稿,1999-03-10接受发表dicated the presence of some xanthones ,polygalitol ,stigmasterol and stigmasterol -3-O -β-glu -copyranoside (Mao et al ,1997,1996;Shbuth et al ,1977).In this paper we describe the isolation of five oleanane triterpenoidal saponins (1～5)from the stem bark of P .arillata and the structure of four new saponins together with a known saponin .RESULTS AND DISCUSSIONSFive triterpenoidal saponins were isolated from the polar part of EtOH extract of P .arillata .One of them was identified as the known polygalasaponin XXXV (5)on the basis of its NMR and FAB -MS spectra ,and comparison with literature data which was isolated from Polygala fallax Hemsl .(Zhang et al ,1996a ).The structure of four novel triterpenoidal saponins ,which named arillatanoside A ～D (1～4),were established by concerted application of NMR and MS studies .Arillatanoside A (1)was obtained as a colorless amorphous po wder .It gave a molecular ion peak at m /z 1236(C 58H 92O 20)in the negative FAB -MS and main fragment ion peaks at m /z 1103[M -132-H ]-,1073[M -162-H ]-,971[M -2×132-H ]-,679[M -2×133-2×145-H ]-.The 1H NMR spectrum of 1showed the presence of seven singlet methyl signals at δ0.76,0.85,1.10,1.47,1.51,1.63and 1.90;a pair of hydroxymethyl signals at δ3.58and 3.95;a trisubstituted olefinic proton signal at δ5.79(s ,br .);and five anomeric pr oton signals at δ6.45(s ,br .),6.00(s ,br .),5.12(s ,br .),5.05(s ,br .)and 5.02(s ,br .).The 13C NMR spectrum of 1showed the presence of one carboxylic carbon signal at δ182.15,one ester carbonyl carbon signal at δ176.75and five anomeric carbon signals at δ106.84,105.86,105.30,101.08and 94.89.The13C and 1H NMR spectral data of 1were homologous to those of polygalasaponin XXVIII (6),an oleanane triterpenoidal saponin whic h isolated from Polygala japonica Houtt .(Zhang et al ,1996b ;Masayuki et al ,1995).The car bon signals for aglyc one skeleton and sugar moiety of 1were very simi -lar to those of 6(Table 1).It is indicated that both of them have the same a glycone as presenegenin and similar sugar linkages .Ho wever ,in the comparison between the 13C NMR spectrum of 1and those of 6,the spectrum of 1showed one set additional signals of α-L -arabinopyranosyl unit .A careful analysis of the glycosylation shift led us obser ved that the signal C -3of terminal β-D -xylopyranosyl unit of oligosaccharide chain of 1was do wnfield shifted to δ87.79from δ78.8of 6,while other car -bon signals were almost unaffected .It was suggested that the additional α-L -arabinopyranosyl unit of 1could be linked to C -3position of the terminal β-D -xylopyranosyl unit of 6.This was confirmed by two -dimensional NMR techniques .H MQC and HMBC experiments showed c orrelation between H -3of β-D -xylopyranosyl unit and C -1of α-L -arabinopyranosyl unit .Based on the above evi -dence ,the structure of saponin 1was established to be 28-O -α-L -arabinopyranosyl -(1※3)-β-D -xylopyranosyl -(1※4)-α-L -rhamnopyranosyl -(1※2)-β-D -fucopyranosyl pr esene -genin -3-O -β-D -gluc opyranoside .Arillatanoside B (2)was obtained as a white amorphous powder and exhibited a molecular ion358 云 南 植 物 研 究 21卷peak at m /z 1440by negative FAB -MS .To comparison with 13C NMR spectrum suggested its molec -ular formula could be C 66H 104O 34.The 13C NMR spectrum of 2showed the presence of one car boxylic carbon signal at δ185.91,two ester carbonyl carbon signals at δ176.07and 171.25,and six anomeric carbon signals at δ106.64,105.39(2×C ),103.25,102.23and 94.57.It is noticed that the 13C NMR spectrum of 2closely resembled that of polygalasaponin XXXIV (7)(Zhang et al ,1996)except one more α-L -arabinopyranosyl unit in 2(Table 1).By c omparison of the 13C NMR spectral data of 2with that of 7,all the carbon signals overlapped with each other except for C -3of β-D -xylopyranosyl unit .The chemical shift C -3of β-D -xylopyranosyl unit went downfield from δ76.7in 7to δ87.23in 2,indicated that this additional α-L -arabinopyranosyl unit was located at C -3of β-D -xylopyranosyl unit in 2.Therefore ,the structure of saponin 2was shown to be 28-O -β-D -galactopyranosyl -(1※4)-[α-L -arabinopyranosyl -(1※3)]-β-D -xylopyranosyl -(1※4)-α-L -rhamnopyranosyl -(1※2)-[4-O -acetyl ]-β-D -fuc opyranosyl pr esene -genin -3-O -β-D -gluc opyranoside .Arillatanoside C (3)was obtained as a white amorphous and exhibited a molec ular ion peak atm /z 1398[M (C 64H 102O 33)]-in its negative FAB -MS .The 1H and 13C NMR spectra of 3sho wed six anomeric pr oton signals at δ6.62(s ,br .),6.01(d ,J =8.0Hz ),5.01(s ,br .),4.90(s ,br .),4.78(s ,br .)and 4.78(s ,br .);and six anomeric carbon signals at δ106.55,105.99,105.17,103.16,100.93and 94.87.The 13C NMR spectrum of 3closely resembled that of parison of the 13C NMR spectral data of 3with that of saponin 1,showed that there is one more β-D -galac -topyranosyl unit in 3(Table 1).The C -4carbon signal of β-D -xylopyranosyl unit was downfield shift from δ70.45in 1to δ78.03in 3.It indicated that this additional β-D -galactopyranosyl unit should be linked at the position C -4of β-D -xylopyranosyl unit in 3.Moreover ,the che mical shift pattern of 3are most overlapped with that of saponin 2,except less a set signals of an acetyl gr oup in C -4position of α-L -r hamnopyranosyl unit .Thus ,the structure of saponin 3is 28-O -β-D -galactopyranosyl -(1※4)-[α-L -arabinopyranosyl -(1※3)]-β-D -xylopyranosyl -(1※4)-α-L -rhamnopyranosyl -(1※2)-β-D -fucopyranosyl presenegenin 3-O -β-D -glucopyra -noside .Arillatanoside D (4)exhibited a molecular ion peak at m /z 1530[M (C 69H 110O 37)]-in its neg -ative FAB -MS .The 13C NMR spectr um of 4showed seven anomeric carbon signals at δ111.77,105.08(2×C )104.40,103.27,101.65and 94.56.Its 13C NMR spectrum showed a similar pat -tern to those of saponins 3and desacylsenegasaponin A (8),later was isolated from Polygala senega var .latifolia Torrey et Gray (Masayuki et al ,1995).However ,4exhibited one more α-L -ara -binopyranosyl unit at C -3position of β-D -xylopyranosyl unit in 8,and one mor e β-D -apiofu -rarnosyl unit at C -3position of α-L -r ha mnopyranosyl unit in 3(Table 1).Therefore ,the structure of 4was determined to be 28-O -β-D -galactopyranosyl -(1※4)-[α-L -arabinopyranosyl -(1※3)]-β-D -xylopyranosyl -(1※4)-[β-D -apiofuranosyl -(1※3)]-α-L -rhamnopyra -nosyl -(1※2)-β-D -fucopyranosyl presenegenin -3-O -β-D -glucopyranoside .Though ,the structures of all four new saponins were deduced by comparison with that of kno wn3593期 吴志军等:黄花远志的新齐墩果烷型三萜皂甙 saponins ,as characters ,they are the same aglycone and similar pattern of sugar chain .It is noticed that the triterpenoidal saponins of Polygala species shown biological activity such as inhibitory activity of increasement of serum glucose level has been reported recently (Masayuki et al ,1995and Yoshi -gawa et al ,1999).The screening of biological actives for these ne w saponins is interesting .EXPERIMENTALGeneral experimental procedures 1H and13C NMR spectra were obtained with Bruker AM -400,DR X -500spectrometer .FAB -MS spectra were taken on VG Autospec -3000system spectrometer .The chemical shifts (δ)were expressed in ppm with refer ence to the solvent signals .Coupling c onstants (J )were given in Hz .Chromatographic materials were used Rp -8(40～60μm ,Merck ),Sephadex LH -20(25～100μm ,Pharmacia Fine Chemical Co .Ltd .),MCI -gel C HP20P (75～150μm ,Mitsubish Chemical Industries ,Ltd .)and silica gel (200～300mesh ,Qingdao Marine Chemical Factory ).TLC was developed with CHCl 3-Me OH -H 2O (65∶35∶9,7∶3∶0.5,7∶3∶1).The ratio of solvents was given in v /v in each case .Spot of TLC were detected by spra ying 5%H 2SO 4following by heating .Extraction and isolation Polygala arillata Buch .-Ha m .was collected in Kunming ,Yunnan Province in March 1997.The dried bark (20kg )was po wdered and extracted with 95%EtOH at r oom tempera -ture (4×30L ),then concentrated in vacu -um (60℃)to evaporate the solvent to give a small volume .After extracting with CHCl 3(3×2L ),thewater layer portion was subjected to D101resin column chromatography ,after washing by H 2O ,eluting with EtOH to give 200g polar fraction .150g of them was chromatographed on silica gel column with the solvent system (CHCl 3-MeOH -H 2O ,7∶3∶1)to give fractions A ～F .The fraction A (50g )was rechromatographed over Sephadex L H -20(30%～90%Me OH )and MCI -gel CHP20P (30%～80%Me OH ,30%～50%C H 3COC H 3)to give compound 1(100mg ),2(70mg ),3(80mg ),4(80mg )and 5(90mg )respectively .360 云 南 植 物 研 究 21卷Table 1 13C NMR s pectral data of the aglycone moieties of s aponins (in C 5D 5N )C 6＊7＊5＊8＊12345144.344.344.344.344.5044.2344.4744.3344.29270.370.470.470.170.7470.4067.9770.4570.70386.086.086.086.085.4586.6186.1486.4586.23452.952.552.952.953.5053.4953.4053.2653.38552.552.552.552.652.6052.4652.4152.6852.48621.421.521.521.321.8521.7421.8521.2821.72733.633.633.533.934.1033.6634.0333.9934.00841.241.241.241.241.2641.1841.2341.1541.13949.449.449.349.449.5049.3449.4749.3449.271037.137.037.037.137.0737.0636.8437.1837.001123.623.723.723.723.3023.4223.3323.5023.4312127.9127.9127.8127.8127.94128.28128.12127.80128.2013138.9139.0138.9139.1193.11138.80139.09139.49138.961447.047.047.147.047.0647.2347.0747.0247.221524.624.524.524.524.9524.5924.9424.6424.551624.124.023.924.024.9023.9423.6424.6424.191748.048.148.048.048.2348.2148.4048.1148.171842.042.041.942.042.1542.0342.0941.8141.801945.445.445.445.545.5045.4145.4145.7145.482030.830.830.830.830.8930.8230.7530.8530.752133.833.933.933.934.1034.1034.0333.9933.592232.432.432.432.432.4032.6432.4232.4432.5123180.8180.8180.7180.9182.15185.91185.50186.00186.002414.214.214.214.214.4114.3114.1914.7314.872517.517.517.517.517.6317.7017.6017.5617.532618.818.718.618.818.9118.8018.9419.0818.852764.564.564.464.664.2064.1564.1864.7264.3128176.7176.7176.4176.6176.75176.87176.54176.65176.542933.133.133.033.133.1933.1933.1033.1833.073024.124.023.924.124.0123.9423.8024.0123.88＊ref .dataArillatanoside A (1):The colorless amorphous powder .FAB -MS m /z 1236[M (C 58H 92O 20)]-,1218[M -H 2O ]-,1103[M -132-H ]-,1073[M -162-H ],971[M -2X132-H ]-.1H NMR spectrum :δ0.76,0.85,1.10,1.47,1.51,1.63and 1.90(Me ×7);5.79(1H ,s ,br .,12-H );6.45(1H ,s ,br .),6.00(1H ,s ,br .),5.12(1H ,s ,br .),5.05(1H ,s ,br .)and 5.02(1H ,s ,br .)(anomeric protons ).See 13C NMR data in Table 1and 2.Arillatanoside B (2):The white amorphous powder .FAB -MS :m /z 1440[M (C 66H 104O 34)]-,1308[M -132]-,1278[M -162]-,1145[M -132-163]-,1116[1278-162]-,3613期 吴志军等:黄花远志的新齐墩果烷型三萜皂甙 982[1145-162]-.See 13C NMR data in Table 1and 2.Table 2 13CNMR spectral data of sugar moieties of saponins (in C 5D 5N )C 6＊7＊5＊8＊12345Glu -1105.4105.4105.4105.3105.30105.39105.17105.08105.01275.375.375.375.375.2775.2175.3075.1275.32378.478.378.378.378.2877.5477.8178.5477.86471.671.771.771.471.5771.5071.5071.6571.58578.478.378.378.378.1977.3577.5378.5477.67662.762.862.862.762.6862.3962.5262.7062.58Fuc -194.8943694.294.894.8994.5794.8794.9694.05274.074.173.075.073.5074.3474.5574.8072.50376.774.774.676.376.9074.5176.0076.5874.96473.274.871.273.173.3274.7673.3973.2771.33572.570.670.172.372.5470.7472.4472.4770.27616.916.516.116.917.0216.6116.9217.0016.093-Ac 20.620.64170.1170.124-Ac 20.720.420.9020.43171.1170.8171.25170.84Rha -1101.2101.8102.1101.5101.08102.23100.93101.65102.24271.871.871.471.671.7871.7771.7871.6571.59372.572.572.482.172.5472.6172.6381.9572.50485.185.284.778.785.4185.4986.1578.5484.52568.368.569.068.368.0468.2167.5868.0268.81618.618.818.818.618.5418.9918.9418.8818.67Xyl -1170.4107.0106.8104.8106.84106.64106.55105.08106.70276.275.775.675.176.9075.8375.3076.8275.77378.876.776.676.287.7987.2387.4883.8076.77470.978.378.278.670.4577.7178.0378.1877.67567.565.065.064.667.0066.3066.2865.2164.85Api -1111.7111.77277.677.46379.680.09474.674.80564.665.73Gal -1104.5104.5104.4103.25103.16104.40103.89271.871.871.871.5071.5070.4571.79375.175.175.075.2175.3075.4175.32470.170.070.170.4769.8269.9470.07577.377.377.377.3577.2377.4677.09662.362.362.362.3962.3962.3162.40Ara -1105.86105.39105.99103.27272.5472.6172.6372.47375.4174.7674.5575.62468.8670.2769.8268.02567.3266.6467.1066.64＊ref .Data362 云 南 植 物 研 究 21卷Arillatanoside C (3):The white amorphous .FAB -MS m /z 1398[M (C 64H 102O 33)]-1266[M -132]-,1236[M -162]-.1H NMR spectr um δ6.62(1H ,s ,br .),6.01(1H ,d ,J =8.0Hz ),5.01(1H ,s ,br .),4.90(1H ,s ,br .),4.78(1H ,s ,br .),4.78(1H ,s ,br .)(anomeric pr o -tons ).See 13C NMR data in Table 1and 2.Arillatanoside D (4):The white amorphous .FAB -MS am /z 1530[M (C 69H 110O 37)]-.See13C NMR data in Table 1and 2.Polygalasaponin XXXV (5):The white a morphous po wder .FAB -MS m /z 1349[M (C 63H 98O 31)-H ]-.1H NMR δ2.04(2×CH 3).See 13C NMR data in Table 1and 2.ReferencesJiangsu College of Ne w Medicine ,1979.Dictionary of Traditional Chines e M edicine .Shanghai :Shanghai Science and Technology Pres s ,2071Mao S L ,Liao S X ,Wu J H ,et al ,1996.Studies on chemical c onstituents of Polyga l a arillata Buch -Ham .Acta Phar mac euticaS inica ,31(2):118Mao S L ,Liao S X ,Wu J H et al ,1997.St udies on chemical constituents of Polygala arillata Buch -Ha m .Acta Phar mac euticaS inica ,32(5):360Masayuki Y ,Toshi yuki M ,Takahiro U et al ,1995.Bioactive saponins and gl ycosides .I .Senegae R adix .(1):E -senegasaponins aand b and Z -senegasaponins a and b ,t heir inhibitory effect on alcohol abs orption and hypogl ycemix activity .C hem Pharm Bull ,43(12):2115～2122Shbuth G ,Banerjee S ,Ballava R et al ,1977.Extractives of Polygala ,Part 5.New trioxygenated xanthones of Polygala ar illata .JC hem Soc ,7:740Yos higawa M ,Muraka mo T ,Li Y et al ,1999.Bioactive triterpene glyc os ides from several medicinal plants .In :Yang C R ,Tanaka O(eds .),Advances of Plant Gl ycosides ,Chemis try and Biology ,Elesiver Science ,27～35Zhang D M ,Toshio M ,Mas snori K et al ,1996a .Nine new triterpene s aponins ,polygalas aponin XXXIII -XLI from the roots ofPolyga l a fallax Hemsl .Che m Phar m Bull ,44(11):2092～2099Zhang D M ,Toshio M ,Mas snori K et al ,1996b .Five new triterpene saponins ,polygalas aponin XXVIII -XXXII from the root ofPolyga l a japonica Houtt .Che m Phar m Bull ,44(4):810～8153633期 吴志军等:黄花远志的新齐墩果烷型三萜皂甙 。
环孢素-欧洲药典EP7
Cod-liver oil, farmed
Hale Waihona Puke EUROPEAN PHARMACOPOEIA 7.5
TESTS Acid value (2.5.1) : maximum 2.0. IMPURITIES Anisidine value (2.5.36) : maximum 10.0. Peroxide value (2.5.5, Method B): maximum 5.0. Unsaponifiable matter (2.5.7) : maximum 1.5 per cent, determined on 2.0 g, and extracting with 3 quantities, each of 50 mL, of peroxide-free ether R. Stearin. Heat at least 10 mL to 60-90 °C then allow to cool for 3 h in a bath of iced water or a thermostatically controlled bath A. different ciclosporins [difference from ciclosporin (R = CH3 : at 0 ± 0.5 °C. If necessary, to eliminate insoluble matter, filter ciclosporin A)] : ciclosporin B [7-L-Ala] ; ciclosporin C the sample after heating. The sample remains clear. [7-L-Thr] ; ciclosporin D [7-L-Val] ; ciclosporin E [5-L-Val] ; Positional distribution ((2)-acyl) of fatty acids. Nuclear ciclosporin G [7-(L-2-aminopentanoyl)] ; ciclosporin H magnetic resonance spectrometry (2.2.33). [5-D-MeVal] ; ciclosporin L [R = H] ; ciclosporin T [4-L-Leu] ; Test solution. Dissolve 190-210 mg of the substance to be ciclosporin U [11-L-Leu] ; ciclosporin V [1-L-Abu], examined in 500 μL of deuterated chloroform R. Prepare at least 3 samples and examine within 3 days. Apparatus : high-resolution FT-NMR spectrometer operating at minimum 300 MHz. Acquisition of 13C NMR spectra. The following parameters may be used : — sweep width : 200 ppm (− 5 ppm to 195 ppm) ; B. [6-[(2S,3R,4R)-3-hydroxy-4-methyl-2-(methylamino)octanoic — irradiation frequency offset : 95 ppm ; acid]]ciclosporin A, — time domain : 64 K ; C. isociclosporin A. — pulse delay : 2 s ; — pulse program : zgig 30 (inverse gated, 30° excitation pulse) ; 07/2012:2398 — dummy scans : 4 ; — number of scans : 4096. COD-LIVER OIL, FARMED Processing and plotting. The following parameters may be used : Iecoris aselli domestici oleum — size : 64 K (zero-filling) ; DEFINITION — window multiplication : exponential ; Purified fatty oil obtained from the fresh livers of farmed cod, — Lorentzian broadening factor : 0.2 Hz. Gadus morhua L., solid substances being removed by cooling Use the CDCl3 signal for shift referencing. The shift of the and filtering. central peak of the 1:1:1 triplet is set to 77.16 ppm. Content : Plot the spectral region 171.5-173.5 ppm. Compare the — sum of the contents of EPA and DHA (expressed as spectrum with the spectrum shown in Figure 2398.-1. The shift triglycerides) : 10.0 per cent to 28.0 per cent ; values lie within the ranges given in Table 2398.-1. — vitamin A : 50 IU (15 μg) to 500 IU (150 μg) per gram ; Table 2398.-1. – Shift values — vitamin D3 : maximum 50 IU (1.3 μg) per gram. Signal Shift range (ppm) A suitable antioxidant may be added. PRODUCTION The fish shall only be given feed with a composition that is in accordance with the relevant European Union or other applicable regulations. The content of dioxins and dioxin-like PCBs (polychlorinated biphenyls) is controlled using methods and limits in accordance with the requirements set in the European Union or other applicable regulations. CHARACTERS Appearance : clear, pale yellowish liquid. Solubility : practically insoluble in water, miscible with light petroleum, slightly soluble in ethanol (96 per cent). IDENTIFICATION A. Examine the 13C NMR spectra obtained in the test for positional distribution ((2)-acyl) of fatty acids (see Tests). The spectra contain peaks between 172 ppm and 173 ppm with shifts similar to those in the spectrum shown in Figure 2398.-1. The positional distribution ((2)-acyl) for cervonic (docosahexaenoic) acid (C22:6 n-3 ; DHA), timnodonic (eicosapentaenoic) acid (C20:5 n-3 ; EPA) and moroctic acid (C18:4 n-3) complies with the limits. 4600
13C-NMR
13C
NMR
1st paragraph
(By sensitivity we mean the ease with which signals may be detected under the conditions of measurement.) The nmr signal given by a
13C
NMR
3rd paragraph
This technique involves irradiating the sample with a brief but intense pulse of radio frequency energy, exciting all the 13C nuclei in the sample to their higher spin state. The excited nuclei then relax to their lower energy state, and it is this process that is monitored.
1．灵敏度低 S∝r3· C· P
磁旋比
1H 13C
测定用样品浓度
核的丰度
1H 13C
2.6752 0.6728
99.93% 1.10%
2. 图谱复杂，既有1JC-H，又有2JC-H 和3JC-H， 且1JC-H大
13C
NMR
vocabulary
1st paragraph
• carbon n. [化]碳(元素符号C), (一张)复写纸 carbon paper 复写纸 • Magnetic resonance spectroscopy 核磁共振 • Fluorin n. 氟(元素符号F) [ˈflu(:)ərin] • Phosphor, Phosphorus n. 磷(元素符号P) [ˈfɔsfə] • Nuclei n. [ˈnju:kliai] 核心、中心 [nucleus的复数] [ˈnju:kliəs] nuclear [核]核子的, 原子能的, 核的, 中心的 • isotope n. [化]同位素 isotopic adj. 同位素的 • nuclear spins ：核自旋
第三节 碳核磁共振(13C-NMR)
3 3
即场强相同的情况下，13C核的灵敏度是1H核的 六千分之一，因此，用CW－NMR法不能测13C核。这 也是13C-NMR的发展落后与1H-NMR的原因。 实际上，13C-NMR法发展的历史就是一个不断 克服低灵敏度的历史。 3.提高灵敏度的方法 A.提高磁场强度：随科学技术的进步在不断进行。 B.增加样品中13C核的浓度： 增加样品浓度： 受溶剂溶解度的限制； 用大口径样品管： 在1H谱中用5mm，13C中用 8、10、15mm
一、13C-NMR的发展历史 1957年，瑞典人首先观察到13C-NMR信号； 60年代发现了宽带去偶和付立叶变换技术； 70年代引入脉冲付立叶变换技术和计算机，使 13C-NMR真正进入实用阶段，并在突飞猛进的发展， 成为鉴定有机化合物结构最强有力的手段之一。 目前，NMR领域有两方面显著进展： 硬件方面：H0增强，从 60→90→250→360→750-900MHz 软件方面：二维核磁共振、三维核磁共振； 有了这两方面的进展，直接应用NMR法即可 以确定分子量在5000以下的化合物的结构。
2.远程偶合:间隔2根键以上的偶合 有2JCH(50-60Hz) 、3JCH(10Hz以下)，13C 信号将进一步裂分，形成更为复杂的图形。
六、常见13C－NMR谱的类型及其特征 1.噪音去偶谱（COM） 全氢去偶或13C（1H）宽带去偶（BBD） 在读取13C的FID信号期间，用覆盖所有1H 核共振频率的宽频电磁辐射照射1H核，以消 除所有1H核对相关13C核的偶合影响,大大提 高灵敏度.
富集的13C样品： 用13C合成，成本太 高，一般不用。 C.进行光谱累加：对同一样品进行多次扫 描，用计算机进行累加（CAT）。 D.质子去偶法：消除1H与13C之间的偶合， 使13C峰不分裂，不等价C只有一个峰；此 外，由于NOE效应，信号强度可增加3倍。 E.PFT法：脉冲付立叶变换法是提高灵敏度 的最好方法，也是目前13C-NMR获取的最 主要方法。
pH值对酚醛树脂性能的影响
pH值对酚醛树脂性能的影响高士帅;赵临五;刘美红;陈日清;储富祥【摘要】为了探究pH值对酚醛树脂(PF)理化性能及其压制的胶合板性能的影响,实验分别合成了pH值为10、11、12和13的四种PF树脂,并重点分析了树脂的游离甲醛、游离苯酚、凝胶时间、胶合强度和甲醛释放量的变化规律.通过FT-IR 和13C-NMR对不同pH值的PF树脂进行了结构表征,利用旋转流变仪研究了树脂的固化速度.实验结果表明随着树脂的pH值升高,PF树脂的游离苯酚和游离甲醛的含量均呈下降趋势,凝胶时间先缩短后变长,固化速度先加快再变慢.当pH值为12时,PF树脂和胶合板的性能达到最佳.%The phenol-formaldehyde resin (PF) was synthesized at different pH (10,11,12 and 13) in order to explore the effects of pH on the physical and chemical property of PF and on the property of PF pressed plywood.The change rule of free phenol,free formaldehyde,gel time,bonding strength and formaldehyde emission of PF resins was studied in detail.The structure and curing behavior of PF synthesized at different pH were characterized by using FT-IR and 13C-NMR and rotational rheometer.It is found that the content of free phenol and free formaldehyde of PF decline with increasing of pH,and the gel time of resins initially decreases and then increases.PF prepared at the pH value of 12 shows the best performance of plywood.【期刊名称】《林产工业》【年(卷),期】2017(044)001【总页数】6页(P14-19)【关键词】pH值;酚醛树脂;游离甲醛;固化速度;胶合强度;甲醛释放量【作者】高士帅;赵临五;刘美红;陈日清;储富祥【作者单位】中国林业科学研究院林产化学工业研究所;中国林业科学研究院林产化学工业研究所;中国林业科学研究院林产化学工业研究所;中国林业科学研究院林产化学工业研究所;中国林业科学研究院【正文语种】中文【中图分类】TQ433.4木材工业中使用的酚醛树脂是由甲醛和苯酚在碱性条件下经羟甲基化和缩聚过程合成的热固性甲阶酚醛树脂，具有较好的胶接性能[1]、优良的耐候性、耐沸水性和耐化学腐蚀性，在木材工业中被广泛应用于制造室外级胶合板、集装箱底板等人造板[2,3]。
水煮处理对毛竹物理力学性能的影响
第35卷第3期V ol.35No.32021年5月May2021木材科学与技术Chinese Journal of Wood Science and Technology水煮处理对毛竹物理力学性能的影响李澍农，张亚梅，余养伦，于文吉（中国林业科学研究院木材工业研究所，北京100091）摘要：为了揭示水煮处理对毛竹物理力学性能的影响，研究不同水煮处理时间（4、8、24、48h）对毛竹力学性能、吸水膨胀率和吸水率的影响。
结果表明：毛竹的力学性能随着水煮处理时间的延长而逐渐降低，经过48h水煮处理，静曲强度和弹性模量分别降低了40.24%和43.65%。
经过4h水煮处理，毛竹薄壁组织细胞腔中的淀粉发生糊化，在细胞壁上形成淀粉膜，并堵塞纹孔，同时细胞壁中半纤维素降解、木质素解聚，使得毛竹的吸水率和吸水膨胀率降低；随着水煮处理时间的延长，细胞腔中的淀粉逐渐被降解或溶出，且半纤维素的降解加剧，木质素中的β-O-4键被破坏，从而使竹材的力学性能降低、吸水膨胀率和吸水率增大，造成竹材材质劣化。
关键词：毛竹；水煮处理；静曲强度；弹性模量；吸水膨胀率；吸水率中图分类号：S781.9；S795.9；TS62文献标识码：A文章编号：2096-9694（2021）03-0059-06Effect of Boiling Treatment on Physical and Mechanical Properties ofMoso BambooLI Shu-nong，ZHANG Ya-mei，YU Yang-lun，YU Wen-ji（Research Institute of Wood Industry，Chinese Academy of Forestry，Beijing100091，China）Abstract:To understand the effect of boiling treatment on the physical and mechanical properties of Moso bamboo,the influence of treatment duration(4,8,24,48h)on the mechanical properties,the swelling rate and the water absorption rate of bamboo were studied.The results showed that the mechanical properties of bamboo decreased with increasing treatment durations.The modulus of rupture (MOR)and the modulus of elasticity(MOE)of bamboo with a treatment of48h decreased by40.24%and43.65%respectively.The existing starch on the parenchyma cell wall blocked pits,possibly led to thereduction of the swelling rate and the water absorption rate after the bamboo was treated for4h.Therefore,the dimensional stability of bamboo was improved.However,the starch continued degrading, then,was removed gradually by the hot water,resulting the number of the blocked pits decreased.Meanwhile,the degree of degradation of the hemicellulose increased;and the link ofβ-O-4in the lignin was destroyed.Therefore,the mechanical properties and the dimensional stability of the bamboo decreased if the treatment duration was longer than4h.The quality of the bamboo deterioratedaccordingly.收稿日期：2020-10-29；修改日期：2021-02-09基金项目：国家自然基金青年基金项目“湿热作用下竹材抽提物的形成机制及其对胶合性能的影响机理”（31700482）。
共价功能化POSSPDMS防腐复合涂层的研究
03115段俊等：共价功能化POSS/PDMS防腐复合涂层的研究文章编号：1001-9731（2021）03-03115-07共价功能化POSS/PDMS防腐复合涂层的研究段俊，欧宝立，郭艳（湖南科技大学材料科学与工程学院，湖南湘潭411201）摘要：首先采用硅烷偶联剂通过水解缩合反应方法合成氨基功能化倍半硅氧烷（POSS-NH,）,利用其表面氨基通过表面接枝聚合方法在倍半硅氧烷表面接枝磺化聚苯胺以实现其共价功能化，将磺化聚苯胺功能化倍半硅氧烷和聚二甲基硅氧烷通过溶液共混，利用滴涂法在Q235钢材表面制备出疏水防腐涂层。
通过核磁共振（NMR）和傅里叶转换红外光谱（FT-IR）对氨基功能化倍半硅氧烷和磺化聚苯胺功能化倍半硅氧烷的结构进行表征分析；利用光电子能谱（XPS）分析磺化聚苯胺功能化倍半硅氧烷的化学组成和元素含量比例；利用扫描电子显微镜（SEM）表征磺化聚苯胺功能化倍半硅氧烷的微观形貌，观察到表面形貌较规整、呈颗粒状、纤维较粗的结构；采用静态接触角测试分析了四种涂层的疏水性，制备出的共价功能化POSS/PDMS防腐复合涂层疏水性最佳，接触角达到115°;采用EIS电化学阻抗谱和Tafel极化曲线测试表征了四种涂层的耐腐蚀性能，结果表明：共价功能化POSS/PDMS防腐复合涂层的腐蚀电位更大，腐蚀电流密度较小，由此表明所制备的材料具有优异的防腐性能。
并且发现涂层的接触角越大，防腐性能越好。
关键词：八氨基倍半硅氧烷；磺化聚苯胺；聚二甲基硅氧烷；防腐涂层中图分类号：TG178文献标识码:A0引言腐蚀是通过化学或电化学方法造成金属和合金降解的一种现象，在经济发展、社会安全和个人健康等方面对生活造成了不小的影响，解决金属腐蚀问题是对社会、工业和人民的巨大挑战。
这类问题的解决方案通常是使用隔离层来限制或阻止腐蚀性或氧化性物质从环境到目标的传输。
倍半硅氧烷（POSS, Polyhedral OlgomericSlsesquioxane）作为一种新型的有机/无机杂化纳米材料,由硅、氧元素构成的无机内核和包围在其外围的有机基团R共同组成的笼形化合物，通常可用通式（RSiO"）”表示，R基团可以是活性的，也可以是非活性的，其主要结构包括嵌段、梯形、笼形、半笼形等，分子结构具有很强的可设计性，结构中含有无机的笼型结构和功能化的有机臂,其笼形结构可以通过控制合成条件实现有效的调控。
核磁共振部分习题及答案_2
NMR problems 2nd part1.Below are the 1H and 13C NMR spectra of 2-hexanone (CH3COCH2CH2CH2CH3). Explain carefully how, using homonuclear and heteronuclear decoupling experiments, you could assign the each of the resonances in the 1H and 13C spectra to which nuclei give rise to them.1H NMR spectra of 2-hexanone13C NMR spectra of 2-hexanoneAnswer This is a commonly encountered problem. You obtain a 1H and a 13C NMR spectrum and need to assign each of the resonances in both 1H and 13C spectra.It is generally straight-forward to assign the 1H spectrum using a combination ofdirect inspection (characteristic chemical shifts and multiplicities) in combination with homonuclear decoupling experiments. For 1H spectra, homonuclear decoupling gives you connectivity (which protons are on adjacent carbon atoms) because 1H is 100% abundant. Note that homonuclear decoupling cannot be used for 13C spectra because 13C is isotopically dilute there essentially no possibility that there will be 2 13C nuclei adjacent to each other in an organic molecule.For 2-hexanone, the 1H spectrum contains 5 resonances. You would expect to see one singlet (integral 3H) for the CH3 group attached to the ketone (ie at C1). The CH2 group adjacent to the ketone (ie at C3) would appear as a triplet (with coupling to the adjacent CH2. The CH2 at C4 would appear as a multiplet (a triplet of triplets with coupling to both the CH2 at C3 and the CH2 at C5). Likewise the CH2 at C5 would appear as a multiplet (a triplet of quartets with coupling to both the CH2 at C4 and the CH3 at C6). The resonance of the CH3 at C6 would be a triplet (integral 3H). Intuitively, you would also expect the CH3 and CH2 groups adjacent to the carbonyl to occur at low field (between 1.5 and 2.5 ppm) and the CH3 at C6 to be at high field (between 0.5 and 1 ppm). By inspection, you can assign the proton signals for C1 (δ 1.7), C3(δ 1.9) and C6(δ 0.8). The multiplet signals atδ1.1 andδ 1.3 must belong protons at C4 and C5 and to distinguish these you would use homonuclear decoupling experiments. If the protons at C3 (δ 1.9) are irradiated, the multiplet due to protons at C4 would collapse from a triplet of triplets to a triplet and hence its shift would be known. Similarly, if the protons at C6 (δ 0.8) were irradiated, the multiplet due to protons at C5 would collapse from a triplet of quartets to a triplet and hence its shift would be known.Having assigned the 1H spectrum, the 13C spectrum can be assigned using selective heteronuclear decoupling experiments.The 13C spectrum contains 6 resonances, the resonance due to the carbonyl carbon is obvious from its shift. For the remaining 5 carbons you would expect to have signals from 2 x CH3 and 3 x CH2 groups and in the absence of any 1H decoupling these would appear as 2 quartets and 3 triplets. In the heteronuclear decoupling experiment you would irradiate each of the resonances in the 1H spectrum and observe the 13C spectrum. As each of the 1H signals is irradiated, the resonance of the 13C coupled to it would collapse to a singlet - the multiplicity of the other signals would remain essentially unchanged.The correlation of 1H and 13C NMR spectra can also be achieved using two-dimensional NMR using a heteronuclear shift correlation (HSC) experiment.2.The simple molecules below contain the NMR-active nuclei 14N (I=1), 31P (I=1/2), 1H (I=1/2) and 13C (I=1/2). Considering only the coupling constants through one bond (1J ax) as being significant, construct diagrams which schematically represent the splitting pattern you would see in :1The 1H NMR spectrum of the ammonium ion [N H4+].2The 14N NMR spectrum of the [N H4+].3The 13C NMR spectrum of trimethylamine [(C H3)3N]4The 31P NMR spectrum of phosphine [P H3].5The 1H NMR spectrum of phosphine [P H3].6The 13C NMR spectrum of H2P-C H2-PH2.7The 13C NMR spectrum of [(PH2)3C H].Answer This is simply an exercise in predicting the multiplicity observed using the formula: multiplicity = 2nI + 1.1The 1H NMR spectrum of CH4 will be a singlet (no multiplicity) since the molecule is tetrahedral in shape and all of the protons are equivalent. There is no coupling from carbon since almost all C is 12C and this is NMR silent.2The 13C NMR spectrum of CH4 is a quintet. The C is coupled to 4 equivalent protons and the spin of 1H is ½.(2nI + 1) = (2 x 4 x ½) + 1 = 53The 1H NMR spectrum of NH4+ will have 3 lines. The molecule is tetrahedral in shape and all of the protons are equivalent and these will be coupled to 14N (which has a spin I = 1).(2nI + 1) = (2 x 1 x 1) + 1 = 34The 14N NMR spectrum of NH4+ is a quintet. The 14N is coupled to 4 equivalent protons and the spin of 1H is ½.(2nI + 1) = (2 x 4 x ½) + 1 = 55The 1H NMR spectrum of PH3 is a doublet. The molecule (like ammonia) is pyramidal and all 3 protons are equivalent. The protons (with spin I = ½) are coupled to one 31P nucleus.(2nI + 1) = (2 x 1 x ½) + 1 = 26The 31P NMR spectrum of PH3 will have 4 lines (quartet). The molecule is pyramidal in shape and all of the protons are equivalent (with a spin I = ½). and these will be coupled to 31P.(2nI + 1) = (2 x 3 x ½) + 1 = 47The 13C NMR spectrum of H2PCH2PH2 will be a triplet of triplets. The 13C will be coupled to 2 x 31P (giving a triplet splitting) and the 13C will be coupled to 2 x 1H (giving a triplet splitting).8The 13C NMR spectrum of (H2P)3CH will be a doublet of quartets. The 13C will be coupled to 3 x 31P (giving a quartet splitting) and the 13C will be coupled to 1 x 1H (giving a doublet splitting).3 The 1H NMR spectrum of (E)-2-pentenal (given below) has 5 distinct resonances: δ9.5 (1H, doublet), δ 6.9 (1h, doublet of triplets), δ 6.0 (1H, doublet of doublets), δ 2.2 (2H, multiplet) and δ 1.0 (3H, triplet). Sketch and clearly describe the 1H spectra you would obtain while applying strong Rf. irradiation at :1δ9.52δ 6.93δ 6.04δ 2.25δ 2.26δ 1.0Answer The signals in the spectrum can be readily assigned by inspection: •δ 9.5 is due to the aldehydic proton - it appears as a doublet due to coupling with the vinylic proton on the adjacent carbon.•δ 6.0 is due to the vinyl proton at C2. It appears as a doublet of doublets with one doublet splitting due to the aldehyde proton and the other doublet splitting due to the vinylic proton at C3.•δ 6.9 is due to the vinyl proton at C3. It appears as a doublet of triplets with the doublet splitting due to the vinylic proton at C2 and the triplet splitting due to coupling to the CH2 group at C4.•δ 2.2 is due to the CH2 group at C4. It appears as a multiplet but must be adoublet of quartets with the doublet splitting due to coupling to the vinylic proton at C3 and the quartet splitting due to the CH3 group at C5.•δ 1.0 is due to the CH3 group at C5. It appears as a triplet due to coupling to the CH2 group at C4 (δ 2.2).The effect of irradiation at each of these frequencies is to decouple the nuclei which have signals at these frequencies. Decoupling effectively removes the nucleus from the spin system.1 With irradiation at δ 9.5, the multiplicity of the vinylic CH resonance at δ 6.0is simplified from a doublet of doublets to a doublet. The remaining resonances in the spectrum are unchanged.2With irradiation at δ 6.9, the multiplicity of the vinylic CH resonance at δ 6.0 is simplified from a doublet of doublets to a doublet and the CH2 resonance at C4 (δ 2.2) is simplified from a doublet of quartet to a simple quartet. The remaining resonances are unchanged.3With irradiation at δ 6.0, the multiplicity of the vinylic CH resonance at δ 6.9 is simplified from a doublet of triplets to a simple triplet and the resonance of the aldehyde proton is simplified from a doublet to a singlet.4With irradiation at δ 2.2, the multiplicity of the vinylic CH resonance δ 6.9 is simplified from a doublet of triplets to a simple doublet and the multiplicity of the CH3 resonance (δ 1.0) is simplified from a triplet to a singlet.5With irradiation at δ 1.0, the multiplicity of the CH2 group at C4 (δ 2.2) is simplified from a doublet of quartets to a simple doublet. The remaining resonances in the spectrum are unchanged.4.The 1H NMR spectrum of 1,4-dioxan-d7 at room temperature consists of a singlet (with broadband decoupling of 2H). At -150o C the spectrum appears as two singlets of equal intensity. Using your knowledge about the conformation (and conformational mobility) of 6-membered rings, give a rationalisation as to why the room temperature and low temperature spectra appear as they do.Answer 1,4-dioxan is a 6-membered ring and adopts a chair conformation (like other 6-membered rings such as cyclohexane). In the chair conformation, there are two proton environments and protons can be either axial or equatorial. At room temperature, rapid ring flipping causes protons to exchange between axial and equatorial sites so the proton spectrum is a singlet.In 1,4-dioxan-d7, seven out of the 8 protons have been substituted deuterium, leaving only 1 proton. Again this proton would be expected to be in either the axial or equatorial position and at low temperature (slow exchange) the protons in axial positions will have a different chemical shift to protons in the equatorial sites.At low temperature where the ring flip is slow, there will be molecules with the single proton axial some with the proton equatorial. There would be two signals one corresponding to axial protons and one corresponding to equatorial protons. The fact that these signals are of approximately equal in intensity indicates that there is no significant thermodynamic preference to have the proton to be equatorial or axial.At high temperature, the ring flip is rapid and the proton is rapidly exchanged between axial and equatorial sites and the shift is averaged to a single resonance at a shift half way between the signals between the signals in the slow exchange spectrum.5.The 1H free induction decay (FID) was acquired following a single 90o pulse to a sample of chloroform (CHCl3). The spectrum following Fourier transformation (FT) is given on the right.Describe what would happen to the FID and to the spectrum obtained following FT when :1The pulse angle was increased to 150o.2 A small amount of an Fe(III) salt was dissolved in the solution before acquisitionof the FID.316 FIDs were accumulated and added together before FT.4 A second FID was acquired using a 270o pulse instead of a 90o pulse and added tothe first before FT.5The sample was dissolved in a viscous solvent before acquisition of the FID.6The FID was accumulated for twice the acquisition time before FT.7The proton spectrum was saturated for several seconds immediately before (but not during) the acquisition of the FID.What would happen to the spectrum if 16 FIDs were accumulated, each wastransformed and the resulting spectra were added together to give the final spectrum ? Answer The 1H FID and spectrum are derived from a sample of CHCl3.1The response of the sample is a sinusoidal function of the pulse angle. Sin(90) = 1 and Sin(150) = Sin(30) = ½ so the intensity of the signal in the FID and in the spectrum will be decreased by a factor of 2.2Addition of a small amount of a paramagnetic salt will increase the efficiency of relaxation. Nuclei would be expected to relax more quickly and this would be reflected in a change in the appearance of the FID and the spectrum. The signal in the FID would decay to zero more rapidly (i.e. in less time); signals in the spectrum would be broader.3The signal-to-noise ratio (S/N) of the spectrum increases as the square root of the number of acquisitions that are added together. If 16 acquisitions are added the S/N increases by a factor of √ 16 = 4.4If an FID was accumulated with a 270o pulse, it would be a mirror image (about the zero level in the FID) of the FID accumulated with a 90o pulse. This means that at every point where the 90o FID was above zero, the 270o FID would be below zero and addition of the 90o and 270o FID's would mean that the signals would exactly cancel - only noise would remain. The spectrum would contain only noise.5 A viscous solvent will slow the rate of molecular tumbling and this in turnincreases the efficiency of relaxation. Nuclei would be expected to relax more quickly and this would be reflected in a change in the appearance of the FID and the spectrum. The signal in the FID would decay to zero more rapidly (ie in less time); signals in the spectrum would be broader.6The FID as presented has clearly decayed to zero well before the accumulation of the FID has stopped. By the end of the acquisition time, there is no signal only noise being accumulated. Increasing the acquisition time by a factor of two will double the time for which the FID signal is accumulated however the real signal in the FID will still decay at the same rate so the real signal will be proportionally less of the FID and there would be proportionally more noise. In the spectrum, the S/N would be decreased.7Saturation of the 1H spectrum would mean that the 1H spectrum would not be observable, providing there was no significant delay between the saturation and recording the FID. Nuclei take several T1's to recover following saturation.8If 16 FIDs were accumulated then each transformed and the spectra added, there would be no difference to the situation where 16 FID's were added and the summed FID was transformed. In practice since the FT actually takes some time it is more time-economical to minimise the number of tiems that the FT has to be performed.6.The 13C NMR spectrum of 2-methyl-4,5-dihydrofuran (A) is given below. The spectrum was the result of 32 scans with 1 sec. delay between acquisitions and no proton decoupling.1What would happen to the spectrum if 128 scans were accumulated and added together ?2What would happen to the spectrum if the spectrum was acquired with a long delay (say 300 seconds) between acquisitions?3What would happen to the spectrum if a small amount of soluble paramagnetic salt was added to the sample prior to acquisition ?4What would happen to the spectrum if the 1H spectrum was irradiated with broad-band Rf radiation before (but not during) each acquisition ?5Could the 1H NMR spectrum of this molecule be analysed by first order splitting rules? Why?Answer The 13C spectrum of the compound (A) contains 5 signals and from the multiplicity these can be assigned to the two CH2 carbons, one CH carbon, one CH3 signal and one quaternary carbon.1The signal-to-noise ratio (S/N) of the spectrum increases as the square root of the number of acquisitions that are added together. If 128 acquisitions are added, this4 times the number of scans as the 32 used to accumulate the basic spectrum. TheS/N increases by a factor of √ 4 = 2.2With only a 1 sec delay between acquisitions, it very likely that all of the carbons in the sample are not fully relaxed between scans and hence the relative intensities of the signals are not correct. A long delay between acquisitions would ensure that all the nuclei are fully relaxed and therefore all 5 signals should all be of the same intensity.3 A paramagnetic species added to the sample will cause more efficient relaxationof all nuclei. Any differences in intensity resulting from differences in relaxation times would be reduced. All of the lines in the spectrum would be broadened.4Irradiation of the proton spectrum before (but not during) the acquisition would provide an NOE enhancement to the carbon spectrum (without decoupling carbon from protons). So the intensity of the signals from protonated carbons in the sample would be increased. The spectrum would not be proton decoupled so the multiplicities of signals would remain the same.5There are 8 protons in this molecule and the spin system can be assigned as an AA'GG'RX3 spin system. Because the spin system contains nuclei which arechemically equivalent but magnetically non equivalent it cannot be analysed by first order rules.7.The 1H free induction decay (FID) below was acquired following a single 45o pulse to a sample of containing two species. The spectrum obtained following Fourier transformation (FT) is given on the right.Describe what would happen to the FID and to the spectrum obtained following FT when:1The pulse angle was doubled.2The pulse angle was increased by a factor of 4.3The pulse angle was increased by a factor of 3.464 FIDs were accumulated and added together before FT.Answer The FID and the spectrum are derived from a sample containing 2 species. The spectrum clearly contains one sharp resonance and one broad resonance and you would predict that the broad signal arises from a species with a short relaxation time and the narrow resonance arises from a species with a long relaxation time.1The pulse angle is 45o so increasing this by a factor of 2 will mean that a 90o pulse is delivered to the sample. The response of the sample is a sinusoidal function of the pulse angle. Sin(45) = 1/√ 2 and Sin(90) = 1 so the intensity of the signal in the FID and in the spectrum will be increased by a factor of √ 2 (1.414).2The pulse angle is 45o so increasing this by a factor of 4 will mean that a 180o pulse is delivered to the sample. The response of the sample is a sinusoidal function of the pulse angle. Sin(180) = 0 so the intensity of the signal in the FID and in the spectrum will be zero. Both the FID and the spectrum would contain only noise.3The pulse angle is 45o so increasing this by a factor of 3 will mean that a 135o pulse is delivered to the sample. The response of the sample is a sinusoidal function of the pulse angle. Sin(135) = Sin(45) = 1/ 2 so the intensity of the signal in the FID and in the spectrum will be unchanged.4The signal-to-noise ratio (S/N) of the spectrum increases as the square root of the number of acquisitions that are added together. If 64 acquisitions are added the S/N increases by a factor of √ 64 = 8.。