不同产地六月霜挥发油化学成分比较研究

采用水蒸气蒸馏法提取,并用GC-MS-计算机联用技术对六月霜不同产地挥发油成分进行定性和定量分析.共分离出60余种化合物,鉴定出32种,其主要成分为樟脑、7-二甲氨基-4-甲基-2H.1-苯骈吡喃-2-酮和3-亚甲基-1,7,7-三甲基双环[2,2,1]庚烷-2-甲基-环戊烷甲酸酯.不同产地六月霜挥发油化学成分种类和含量大致相同.     

华山松木蠹象聚集信息素分离鉴定和引诱效果

为了研制华山松木蠹象Pissodes punctatus Langor et Zhang的引诱剂,对华山松木蠹象的虫粪和雄虫后肠挥发性物质进行了分析鉴定和室内外生物活性测试。经GC-MS测定,发现在华山松木蠹象的新鲜虫粪和雄虫后肠挥发性物质中,除了松树挥发性的单萜烯如α-蒎烯、β-蒎烯和3-蒈烯以外,还存在1-甲基-2-异丙烯基-环丁烷乙醇（grandisol）。室内Y-型嗅觉仪趋向实验表明,较低浓度的3-（+）-蒈烯、1-甲基-2-异丙烯基-环丁烷乙醇及其相应的醛1-甲基-2-异丙烯基-环丁烷2醛（grandisal）,引起华山松木蠹象的正趋向反应。林间引诱试验表明,1-甲基-2-异丙烯基-环丁烷乙醇和1-甲基2-异丙烯基-环丁烷2醛对华山松木蠹象具有一定的引诱作用。由此推断,1-甲基-2-异丙烯基-环丁烷乙醇可能是华山松木蠹象的集结信息素成分之一。     

木荷花挥发性成分的GC-MS分析

采用无水乙醚超声萃取得到新鲜木荷(Schima superba)花浸膏提取物,顶空固相微萃取富集挥发性成分,气相色谱-质谱联用仪分析,归一化法计算各组分的相对含量.鉴定出挥发性化合物中的51个成分,约占相对总含量的99%;挥发性成分中含氧化合物的含量超过93%,其中主要的化合物及其相对含量为酮代异佛尔酮(26.33%)、氧化芳樟醇(19.53%)、环氧芳樟醇(8.80%)、3,7-二甲基-2,6-辛二烯-1-醇(8.23%)、白藜芦素(7.89%)、4-羟基3,5,5-三甲基-2-环己烯-1-酮(6.54%)、2,6,6-三甲基-1,4-环己二酮(4.06%)、苯乙醇(2.17%)、2-甲基-2-壬烯-1-醇(2.04%)等.     

动物气味的顶空取样技术及其在水貂肛腺成分分析中的应用

我们设计了一种用于动物气味的顶空 (headspace)取样装置 ,并结合溶剂解吸附和气质联用技术 ,通过对水貂 (Mustelavision)肛腺分泌物的挥发性成分的分析验证这种装置的可行性。出现 5个气谱峰 ,对应的质谱显示其分子量和分子式依次为峰 1:10 2 (C5H10 S)、峰 2 :10 4(C5H12 S)、峰 3:10 2 (C5H10 S)、峰 4:136(C5H12 S2 )和峰 5 :134 (C5H10 S2 )。峰 1、峰 3和峰 5分别为以前研究所鉴定出的 3种主要成分 :2 ,2 二甲基硫代环丁烷、2 乙基硫代环丁烷和 3,3 二甲基 1,2 二硫代环戊烷 ,2 ,2 二甲基硫代环丁烷为优势成分 ,组成无性别间差异。这些结果与以前研究一致 ,说明了这种顶空取样装置的可靠性。峰 2和峰 4两种成分以前在水貂肛腺中未曾发现 ,初步分析可能分别为 3 甲基 1 丙基硫醇和 1,5 戊二硫醇。尽管这两个成分有待进一步分析确定 ,但可以肯定分子量为 10 4和 136的成分不仅在水貂肛腺分泌物中没有发现过 ,而且在已经研究过的鼬属动物和食肉类的肛腺中都未发现过 ,能检测出更多的成分 ,说明这种方法的可靠性很高。     

黄鼬肛腺分泌物的挥发性成分

利用顶空取样、溶剂解吸附和气质联用技术分析了黄鼬（Mustela sibirica)肛腺分泌物的挥发性成分。鉴定出的六种化合物均为含硫的环状有机物：（1）2,2－二甲基硫代环丁烷;（2）顺或反2,4－二甲基硫代环丁烷;（3）反－2,3－二甲基硫代环丁烷;（4）2－乙基硫代环丁烷;（5）2－丙基硫代环丁烷;（6）3,3－二甲基－1,2－二硫代环戊烷。尽管黄鼬肛腺成分的组成和鼬鼠其它种存在很大的相似性,但是成分组成的种间差异很明显。另外,2－乙基硫代环丁烷仅存在于雌性黄鼬中。很多研究已经证明对鼠类有驱赶作用的2,2－二甲基硫代环丁烷和2－丙基硫代环丁烷在黄鼬肛腺分泌物同时存在,说明黄鼬肛腺分泌物对鼠类可能有很强的驱避作用。     

中红侧沟茧蜂化学感受蛋白MmedCSP1的结合特征

化学感受蛋白是一类存在于昆虫化学感受器中的可溶性蛋白, 被认为与昆虫识别外界化学信息有关。本研究使用pGEX-4T-1表达载体在BL21 (DE3)异源表达系统中表达中红侧沟茧蜂Microplitis mediator化学感受蛋白MmedCSP1, 并通过亲和层析法纯化得到去表达标签的MmedCSP1;使用bis-ANS作为荧光配基, 在荧光分光光度计上研究它与50种气味标样的结合特征, 从而得到此类化学感受蛋白在中红侧沟茧蜂嗅觉识别中识别气味的种类。结果表明: MmedCSP1只能与水杨酸甲酯、戊烷、罗勒烯、β-紫罗兰酮、3, 4-二甲基苯甲醛、2-己酮和叶醇结合。但只有脂类化合物β-紫罗兰酮能在浓度为1 mmol/L下将bis-ANS从MmedCSP1中替换50%, β-紫罗兰酮与MmedCSP1的结合常数为16.89 μmol/L。这些结果提示MmedCSP1参与中红侧沟茧蜂对水杨酸甲酯、戊烷、罗勒烯、β-紫罗兰酮、3, 4-二甲基苯甲醛、2-己酮和叶醇等气味的识别过程, 且在不同气味中的识别过程中对于气味的运输能力有差异。     

动物气味的项空取样技术及其在水貂肛腺成分分析中的应用

我们设计了一种用于动物气味的顶空（heqadspace）取样装置,并结合溶剂解吸附和气质联用技术,通过对水貂（Mustela vision）肛腺分泌物的挥发性成分的分析验证这种装置的可行性,出现5个气谱峰,对应的质谱显示其分子量和分子式依次为峰1：102（C5H10S）、峰2：104（C5H12S）、峰3：102（C5H10S）、峰4：136（C5H12S2）和峰5：134（C5H10S2）。峰1、峰3和峰5分别为以前研究所鉴定出的3种主要成分：2,2－二甲基硫代环本烷、2－乙基硫代环丁烷和3,3－二甲基－1,2－二硫代环戊烷,2,2－二甲基硫代环丁烷为优势成分,组成无性别间差异。这些结果与以前研究一致。说明了这种顶空取样装置的可靠性,峰2－峰4两种成分以前在水貂肛腺中未曾发现,初步分析可能分别为3－甲基－1－丙基硫醇和1,5－戊二硫醇,尽管这两个成分有待进一步分析确定,但可以肯定分子量为104和136的成分不仅在水貂肛腺分泌物中没有发现过,而且在已经研究过的鼬属动物和食肉类的肛腺中都未发现珲,能检测出更多的成分,说明这种方法的可靠性很高。     

白木香内生真菌Botryosphae riarhodina A13固体发酵产物的分离鉴定

运用硅胶柱、反相硅胶柱、凝胶柱、HPLC和薄层制备等色谱技术及重结晶从白木香内生真菌Botryosphaeria rhodina A13的固体发酵产物中分离纯化了10个化学成分,通过化合物波谱数据及理化性质鉴定它们的结构分别为豆甾-4-烯-3-酮(1)、豆甾-5-烯-3β-醇-7-酮(2)、豆甾-4-烯-3,6-二酮(3)、(22E,24S)-5α,6α-环氧基-24-甲基胆甾-8(14),22-二烯-3β,7α-二醇(4)、5,4'-二羟-7-甲氧基黄酮(5)、香草酸(6)、对甲氧基苯甲酸(7)、尿嘧啶(8)、2-甲氧基对苯二酚(9)、2-甲基-3,5-羟基色酮(10)。其中化合物1~10均为首次从该属真菌中分离得到,化合物4具有细胞毒活性。     

相思子化学成分研究

本研究从相思子种子的乙醇提取物中首次分离得到8个化合物,通过理化性质及光谱分析鉴定其结构为N-9-甲基-β-咔啉(1)、异喹啉酮(2)、吲哚-3-羧酸(3)、2,3-二甲氧基-5,7-二羟基-二氢黄酮(4)、3-羟甲基呋喃醛(5)、3-羟基-2-甲基-4-吡喃酮(6)、豆甾醇-4,22-二烯-3-酮(7)和(6E,6’E)-2-hydroxypropane-1,3-diylbis(octadec-6-enoate)(8)。同时还得到13个其他成分。     

杠柳根皮挥发油化学成分及对麦二叉蚜的毒杀活性初探

采用GC/M S联用技术,对杠柳根皮中挥发油的化学成分进行了分析,共分离出15种化合物,对其中10种进行了鉴定,已鉴定成分占挥发油总成分的94.91%,分别为4-甲氧基水杨醛、乙酸丁酯、2-甲基-1,3-二氧环戊基乙酸乙酯、乙羟硫代羧酸-8-二乙氧基磷酰正辛酯、1,1,3,3-四丁氧基-2-丙酮、4-甲基-2-戊酮、甲酸丁酯、1-(1-乙氧基乙氧基)丁烷、2-正丁氧基四氢吡喃及丁醚。以麦二叉蚜为试虫进行挥发油杀虫活性测定。结果表明,杠柳根皮挥发油对试虫具有较强的触杀活性。     

A Glutathione S-Transferase with Activity towards cis-1,2-Dichloroepoxyethane Is Involved in Isoprene Utilization by Rhodococcus sp. Strain AD45

Rhodococcus sp. strain AD45 was isolated from an enrichment culture on isoprene (2-methyl-1,3-butadiene). Isoprene-grown cells of strain AD45 oxidized isoprene to 3,4-epoxy-3-methyl-1-butene, cis-1,2-dichloroethene to cis-1,2-dichloroepoxyethane, and trans-1,2-dichloroethene to trans-1,2-dichloroepoxyethane. Isoprene-grown cells also degraded cis-1,2-dichloroepoxyethane and trans-1,2-dichloroepoxyethane. All organic chlorine was liberated as chloride during degradation of cis-1,2-dichloroepoxyethane. A glutathione (GSH)-dependent activity towards 3,4-epoxy-3-methyl-1-butene, epoxypropane, cis-1,2-dichloroepoxyethane, and trans-1,2-dichloroepoxyethane was detected in cell extracts of cultures grown on isoprene and 3,4-epoxy-3-methyl-1-butene. The epoxide-degrading activity of strain AD45 was irreversibly lost upon incubation of cells with 1,2-epoxyhexane. A conjugate of GSH and 1,2-epoxyhexane was detected in cell extracts of cells exposed to 1,2-epoxyhexane, indicating that GSH is the physiological cofactor of the epoxide-transforming activity. The results indicate that a GSH S-transferase is involved in the metabolism of isoprene and that the enzyme can detoxify reactive epoxides produced by monooxygenation of chlorinated ethenes.     

Characterization of the gene cluster involved in isoprene metabolism in Rhodococcus sp. strain AD45

The genes involved in isoprene (2-methyl-1,3-butadiene) utilization in Rhodococcus sp. strain AD45 were cloned and characterized. Sequence analysis of an 8.5-kb DNA fragment showed the presence of 10 genes of which 2 encoded enzymes which were previously found to be involved in isoprene degradation: a glutathione S-transferase with activity towards 1,2-epoxy-2-methyl-3-butene (isoI) and a 1-hydroxy-2-glutathionyl-2-methyl-3-butene dehydrogenase (isoH). Furthermore, a gene encoding a second glutathione S-transferase was identified (isoJ). The isoJ gene was overexpressed in Escherichia coli and was found to have activity with 1-chloro-2,4-dinitrobenzene and 3,4-dichloro-1-nitrobenzene but not with 1, 2-epoxy-2-methyl-3-butene. Downstream of isoJ, six genes (isoABCDEF) were found; these genes encoded a putative alkene monooxygenase that showed high similarity to components of the alkene monooxygenase from Xanthobacter sp. strain Py2 and other multicomponent monooxygenases. The deduced amino acid sequence encoded by an additional gene (isoG) showed significant similarity with that of alpha-methylacyl-coenzyme A racemase. The results are in agreement with a catabolic route for isoprene involving epoxidation by a monooxygenase, conjugation to glutathione, and oxidation of the hydroxyl group to a carboxylate. Metabolism may proceed by fatty acid oxidation after removal of glutathione by a still-unknown mechanism.     

Hemoglobin adducts and micronuclei in rodents after treatment with isoprene monoxide or butadiene monoxide

1,3-Butadiene and isoprene (2-methyl-1,3-butadiene) are chemically related substances that are carcinogenic to rodents. The overall aim of this work is to elucidate the role of the genotoxic action of diepoxide metabolites in the carcinogenesis of the dialkenes. In vivo doses of the diepoxide metabolites were measured through reaction products with hemoglobin (Hb adducts) in studies of induced micronuclei (MN) in rodents. In the reaction with N-terminal valine in Hb, diepoxybutane and isoprenediepoxide form ring-closed adducts, pyrrolidines [N,N-(2,3-dihydroxy-1,4-butadiyl)valine and N,N-(2,3-dihydroxy-2-methyl-1,4-butadiyl)valine, respectively]. The method applied for Hb-adduct measurement is based on tryptic degradation of the protein and liquid chromatography electrospray ionisation tandem mass spectrometry (LC-ESI-MS/MS) analysis. Mice were given single i.p. injections of the monoepoxides of butadiene and isoprene, 1,2-epoxy-3-butene or 1,2-epoxy-2-methyl-3-butene, respectively. Rats were treated in the same way with 1,2-epoxy-3-butene. In mice pyrrolidine adduct levels increased with increasing administered doses of the monoepoxides. The in vivo dose of diepoxybutane was on average twice as high (0.29+/-0.059 mMh) as the in vivo dose of isoprenediepoxide (0.15+/-0.053 mMh) per administered dose (mmol/kg body weight) of the monoepoxides. In mice the genotoxic effects of the two monoepoxides, measured as the increase in the frequencies of micronuclei (MN), were approximately linearly correlated to the in vivo doses of the diepoxides (except at the highest dose of diepoxybutane). In rats the pyrrolidine-adduct levels from diepoxybutane were below the limit of quantification at all administered doses of 1,2-epoxy-3-butene and no significant increase was observed in the frequency of MN. Measurement of the ring-closed adducts to N-termini in Hb by the applied method permits analysis of in vivo doses of diepoxybutane and isoprenediepoxide, which may be further used for the elucidation of the mechanisms of carcinogenesis of butadiene and isoprene.     

Metabolism of 2-Methylpropene (Isobutylene) by the Aerobic Bacterium Mycobacterium sp. Strain ELW1

An aerobic bacterium (Mycobacterium sp. strain ELW1) that utilizes 2-methylpropene (isobutylene) as a sole source of carbon and energy was isolated and characterized. Strain ELW1 grew on 2-methylpropene (growth rate = 0.05 h⁻¹) with a yield of 0.38 mg (dry weight) mg 2-methylpropene⁻¹. Strain ELW1 also grew more slowly on both cis- and trans-2-butene but did not grow on any other C₂ to C₅ straight-chain, branched, or chlorinated alkenes tested. Resting 2-methylpropene-grown cells consumed ethene, propene, and 1-butene without a lag phase. Epoxyethane accumulated as the only detected product of ethene oxidation. Both alkene consumption and epoxyethane production were fully inhibited in cells exposed to 1-octyne, suggesting that alkene oxidation is initiated by an alkyne-sensitive, epoxide-generating monooxygenase. Kinetic analyses indicated that 1,2-epoxy-2-methylpropane is rapidly consumed during 2-methylpropene degradation, while 2-methyl-2-propen-1-ol is not a significant metabolite of 2-methylpropene catabolism. Degradation of 1,2-epoxy-2-methylpropane by 2-methylpropene-grown cells led to the accumulation and further degradation of 2-methyl-1,2-propanediol and 2-hydroxyisobutyrate, two sequential metabolites previously identified in the aerobic microbial metabolism of methyl tert-butyl ether (MTBE) and tert-butyl alcohol (TBA). Growth of strain ELW1 on 2-methylpropene, 1,2-epoxy-2-methylpropane, 2-methyl-1,2-propanediol, and 2-hydroxyisobutyrate was fully inhibited when cobalt ions were omitted from the growth medium, while growth on 3-hydroxybutyrate and other substrates was unaffected by the absence of added cobalt ions. Our results suggest that, like aerobic MTBE- and TBA-metabolizing bacteria, strain ELW1 utilizes a cobalt/cobalamin-dependent mutase to transform 2-hydroxyisobutyrate. Our results have been interpreted in terms of their impact on our understanding of the microbial metabolism of alkenes and ether oxygenates.     

Stereochemical assignment of naturally occurring 2,3-epoxy-2-methylbutanoate esters

A method to determine the absolute configuration of 2,3-epoxy-2-methylbutanoate ester residues in natural products is presented, based on (i) the reduction of the ester function to yield a 2-methyl-1,2-butanediol, (ii) esterification of the obtained primary alcohol with either (R)-(+)- or (S)-(-)-Mosher's acid to afford the corresponding Mosher's ester, and (iii) 1H-NMR spectral comparison of the final product with that of the Mosher's esters prepared from 2-methyl-1,2-butanediols of known stereochemistry.     

Cordiachromes from Auxemma oncocalyx

Further cordiachromes, rel-10,11β-epoxy-11-ethoxy-8-hydroxy-2-methoxy-8aβ-methyl-5,6,7,8,8a,9,10aβ-octahydro-1,4-anthracendione, 6-formyl-2-methoxy-9-methyl-7,8-dihydro-1,4-phenanthrendione, rel-8,11;9,11-diepoxy-1,4-dihydroxy-2-methoxy-8aβ-methyl-5,6,7,8,8a,9,10,10aβ-octahydro-10-anthracenone, rel-9,11-epoxy-1,4,8-trihydroxy-2-methoxy-8aβ-methyl-5,6,7,8,8a,9,10,10aβ-octahydro-10-anthracenone, rel-2″-methoxy-7″-methyl-1″,4″-naphtalendione-(6″→5)-tetrahydropyran-(2-eq→O→2ax)-tetrahydropyran-(5′→6)- 2-methoxy-7-methyl-1,4-naphthalendione, together with the known, allantoin, sitosterol and 3β-O-d-glucopyranosylsitosterol, have been isolated from Auxemma oncocalyx. Their structures were determined from spectral data, including 2D NMR experiments.     

Pleurotus species convert monoterpenes to furanoterpenoids through 1,4-endoperoxides

Enzymatic synthesis of furanoterpenoids from β-myrcene and related monoterpenes was observed using a solubilised enzyme fraction of mycelium lyophilisates of several Pleurotus species. The initial enzymatic step, the incorporation of molecular oxygen into the conjugated 1,3-diene moiety, was similar to a 2 + 4 cycloaddition of 1,3-dienes with dienophilic ¹O₂, and was followed by a non-catalysed degradation sequence leading to the furans. The cyclic peroxides 3,6-dihydro-4-(2-(3,3-dimethyloxiran-2-yl)ethyl)-1,2-dioxine and 5-(3,6-dihydro-1,2-dioxin-4-yl)-2-methylpentan-2-ol were identified as key intermediates. Biotransformation of β-myrcene in ¹⁸O-labelled HEPES-buffer did not yield a detectable label in perillene, so a water addition to 3,10-epoxy-β-myrcenes as an alternative was ruled out. The pathway suggested presents a substantiated biogenetic scheme for the formation of monoterpenoid furans and opens biotechnological access to valuable natural flavour compounds, such as perillene and rosefurane. Only one metabolite, identified as the new natural compound 6-methyl-2-methylene-hept-5-enal, carried the ¹⁸O-label. The enzymatic formation of this compound through a 1,2-endoperoxide (3-(5-methyl-1-methylene-hex-4-enyl)-[1,2]-dioxetane) is suggested. The label may simply result from a chemical oxygen exchange between the carbonyl group and the ¹⁸O-labelled water.     

Glycogen phosphorylase activity in pharate adults of the stable fly and the effects of a juvenile hormone analogue

Glycogen phosphorylase was assayed in homogenates of pharate adults of the stable fly, Stomoxys calcitrans, by the release of inorganic phosphorous (Pi) from glucose-1-phosphate (G-1-P) in the presence of glycogen. Activity was determined as active and total phosphorylase present by the absence or presence of adenosine-5-monophosphate (AMP) in homogenates. Homogenates of the pharate adults (that were treated topically immediately after larval-pupal apolysis with a synthetic insect juvenile hormone analogue, (E)-4-[(6,7-epoxy-3-ethyl-7-methyl-2-nonenyl)oxy]-1,2-(methylenedioxy)benzene) displayed no inhibition or enhancement of phosphorylase activity when compared to untreated pharate adults.     

A preparation of 3alpha-methyl-5alpha-cholestane-2beta,3beta-diol.

A simple preparation of 3α-methyl-5α-cholestane-2β, 3β-diol (1a) by a four-step synthesis from 2α, 3α-epoxy-5α-cholestane is described.     

The characteristic smell emitted from two scale insects,Ceroplastes japonicus and Ceroplastes rubens

ABSTRACT

The volatile components emitted from two scale insects, Ceroplastes japonicus and Ceroplastes rubens, were identified using GC–MS analysis. The major volatile components of the solvent extract from C. japonicus were α-humulene (35.8%) and δ-cadinene (17.0%), while those of C. rubens were β-selinene (10.3%) and β-elemene (5.1%). In GC/olfactometry, linalool, butyric acid, 3-methylbutyric acid, 2-methylbutyric acid, and vanillin were identified as the odor-active components of the extract from C. japonicus, in addition to trace amounts of trans-4,5-epoxy-(2E)-decenal, 4-methyl-(3E)-hexenoic acid, and phenylacetic acid. With regard to C. rubens, trans-4,5-epoxy-(2E)-decenal, 3-methylbutyric acid, and phenylacetic acid were identified as the odor-active components. Besides, decan-1,4-olide (γ-decalactone) with milky cherry-like note and 3-hydroxy-4,5-dimethylfuran-2(5H)-one (sotolone) with brown sugar-like note were also detected as the characteristic cherry-like sweet-and-sour note of these two scale insects.