利用固相微萃取技术分析锈色粒肩天牛幼虫虫粪所含的挥发物成分

花绒寄甲Dastarcus helophoroides (Fairmaire)成虫主要利用天牛幼虫蛀食树木时排出的虫粪所释放的挥发性化合物找到寄主所在的微栖境.本试验分别应用PDMS,DVB/CAR/PDMS,CAR/PDMS三种萃取头,采用固相微萃取技术对锈色粒肩天牛Apriona swainsoni (Hope)幼虫虫粪的挥发物进行了提取,再经气相色谱-质谱联用仪(GC-MS)进行化合物的鉴定.结果表明,虫粪中的挥发性物质主要为萜烯类,其中α-蒎烯的含量最高,柯巴烯的含量次之.三种萃取头在相同的温度和提取时间下,DVB/CAR/PDMS的萃取头的效果最好.     

黄酒中挥发性和半挥发性成分检测固相微萃取条件的优化

采用固相微萃取-气相色谱联用质谱法分析黄酒中的挥发性和半挥发性成分,对萃取时间、萃取温度、预热时间、酒样添加量、盐添加量、解析时间等条件参数进行了优化,最终确定最佳条件为采用50/30 μm DVB/CAR/PDMS萃取头,在20 mL顶空瓶中,加入6 mL黄酒样和2.0 g NaCl,萃取温度60℃,预热时间15 min,萃取时间30 min,解析温度250℃,解析时间6 min。本方法简单快速,干扰少,灵敏度高。     

糖蜜草挥发性成分HS-SPME-GC-MS分析

本文首次采用顶空固相微萃取技术(HS-SPME)提取该植物挥发性成分,然后通过气质联用技术(GC-MS)结合保留指数以及标样对照对化学成分进行了定性,并通过峰面积归一化法计算出各组分的相对百分含量。试验确定了PDMS/DVB固相微萃取对糖蜜草挥发物的萃取效果最好,并从中鉴定出33种化合物占总挥发性成分的90.94%,主要为烷烃和萜烯类化合物。该研究为开发糖蜜草作为水稻螟虫的驱避植物进行害虫安全有效防治提供理论依据,并为驱避性挥发物的田间应用奠定基础。     

紫锥菊挥发性成分分析研究

采用气-质(GC-MS)联用技术结合顶空固相微萃取(HS-SPME)与传统共水蒸馏法,对紫锥菊植物干花与干根中挥发性成分进行分析方法研究。GC-MS(配N ist2005标准质谱库)从传统方法提取的紫锥菊干根挥发油中分离分析出188个峰,初步鉴定出68种化合物,从聚二甲基硅氧烷(PDMS 7,100μm)及聚丙烯酸酯(PA 85μm)三种固相微萃取涂层吸附的紫锥菊干花挥发性成分中分别分离分析出27、118、105个峰,各自鉴定出1、22、23种化合物。对不同处理方法及紫锥菊植物不同部位所得数据进行了相识性与差异性的比较,所建分析方法及所得结果为了解紫锥菊植物体挥发性有效成分提供了参考。     

三种固相微萃取头对厚皮甜瓜果汁香气成分萃取效果分析

采用美国Supelco公司生产的Carboxen^TM/PDMS、DVB/CAR/PDMS和PDMS/DVB 3种固相微萃取头萃取厚皮甜瓜(Cucumis melo L.)果汁中的香气成分,并用气质联用仪对香气成分进行检测。结果表明, 3种萃取头检测到的香气成分均以酯类为主,醇类和醛类较少。Carboxen^TM/PDMS萃取的香气成分相对含量达91.55%,特征香气成分相对含量达到63.01%;DVB/CAR/PDMS也可以全面萃取甜瓜香气,但香气成分与特征香气的相对含量及总峰面积比Carboxen^TM/PDMS的低;PDMS/DVB萃取头吸附的香气成分种类最少,相对含量最低。可见,对厚皮甜瓜果汁香气成分的萃取以Carboxen^TM/PDMS萃取头较好。     

带鱼初加工过程中风味物质的检测分析

用75μmCAR/PDMS涂层的固相微萃取头萃取带鱼肉中挥发性风味成分,利用气质联用仪分析鉴定各阶段鱼肉中的挥发性成分、种类、相对质量分数等。原料鱼中检出57种化合物,风干6d的鱼肉中检测出61种化合物,以醇类、醛类、酮类、烷烃或烯烃类为主,总质量分数达75.16%。醛类、醇类、酮类、酯类等化合物对鱼肉的风味贡献较大,风味物质的形成与脂类、蛋白质等的降解有关。     

HS-SPME-GC / MS 在大熊猫尿液化学成分检测中的应用

尿液在大熊猫化学通讯过程中具有重要作用。对大熊猫尿液中化学成分的检测是揭示大熊猫尿液中化学物质组成及其功能的关键。本实验通过使用顶空固相微萃取技术(Headspace-solid phase microextraction,HSSPME)对大熊猫尿液样品进行前期处理,继而利用气相色谱- 质谱联用技术(Gas chromatography-mass spectrometry,GC / MS)对大熊猫尿液中化学成分进行检测。共检测到56 个峰,通过在NIST (National Institute of Standards
and Technology)质谱库中进行检索,初步推定出其中的38 种物质。除此之外,还对HS - SPME 萃取头的净化方法进行了探索和改进。结果表明,顶空固相微萃取技术结合气相色谱- 质谱联用技术能够应用于大熊猫尿液中挥发性与半挥发性化合物的检测,并且能够得到较好的实验结果,为揭示大熊猫化学通讯机理提供基础。     

顶空进样与水蒸气蒸馏GC-MS法分析杠板归中挥发性成分

分别采用顶空进样法和水蒸气蒸馏法提取杠板归的挥发性成分,对得到的样品分别进行气相色谱-质谱(GC-MS)分析。对顶空进样得到的挥发性物质和水蒸气蒸馏法得到的挥发油物质进行气质联用色谱分析,在相同条件下用同种检索谱库检索,顶空进样直接得到的挥发性物质进行气质联用色谱分析,得到了42种化合物,含量都大于0.10%,且差别较小;水蒸气蒸馏得到了61种化合物,但含量大于0.10%的只有33种,且差别较大。说明不同的收集方法分别提供了不同沸点和不同状态下杠板归药材中挥发性成分的结构和百分含量;对比综合这两种不同的方法得到的挥发性成分,可得到更全面、更详细的杠板归中挥发性成分的GC-MS实验情况。     

HS-GC/MS法分析乌龙茶挥发性成分

本试验旨在采用静态顶空-气相色谱/质谱法(HS-GC/MS)建立起一种快速测定分析以金观音为代表的乌龙茶中挥发物的方法。本方法利用单因素试验结合L9(34)正交试验,以出峰个数和总峰面积为考察指标,确定了顶空平衡温度、平衡时间、茶/Na Cl以及加样量4种因素的最佳萃取条件,并以此方法对金观音的挥发性成分进行了分析。结果表明,平衡温度对出峰个数和总峰面积的影响最大,而平衡时间的影响最小。试验得出的顶空进样的最优条件为:平衡温度80℃,平衡时间60 min,茶/Na Cl为1∶2,加样量为0. 5 g。在此萃取条件下能够得到104种物质,这些物质能更真实地反映金观音香气成分的化学构成,为以金观音为代表的乌龙茶的开发利用提供有价值的数据。     

荔枝蜜中挥发性成分的指纹图谱研究

蜂蜜中挥发性成分的指纹图谱分析,是科学评价及有效控制蜂蜜质量的重要依据。本文利用顶空固相微萃取-气相色谱-质谱联用法对我国常见5个荔枝品种的30个中意蜂荔枝蜜样品进行挥发性成分分离检测,并结合中药色谱指纹图谱技术建立其指纹图谱。结果表明,不同荔枝品种的荔枝蜜样品其挥发性成分相近,共分离鉴定出41种挥发性成分,建立了由16个共有峰组成的荔枝蜜挥发性成分指纹图谱。另外,发现不同蜂种所酿造荔枝蜜间的挥发性成分存在明显差异,其中1-庚醇、正壬醇和乙酸苯乙酯这3种挥发性成分均仅在中蜂所酿造的荔枝蜜中被检出,而在意蜂所酿造的荔枝蜜中无检出。本文所建立的荔枝蜜挥发性成分指纹图谱,可为荔枝蜜的质量控制提供依据,并为生产上不同蜂种所酿造荔枝蜜的区分提供理论基础。     

Production system influences volatile biomarkers in tomato

Introduction

In recent years, growers have used various production types, including high-tunnel systems, to increase the yield of tomatoes (Lycopersicon esculentum). However, the effect of high-tunnel cultivation, in comparison to conventional open-field production, on aroma and flavor volatiles is not fully understood.

Objectives

To optimize the extraction and quantification conditions for the analysis of tomato volatiles using headspace solid phase microextraction (HS-SPME) coupled with gas chromatography–mass spectrometry (GC–MS), and study the effect of production systems on volatile profiles using metabolomics approach.

Methods

The HS-SPME conditions were optimized for extraction and GC–MS was used to quantify the volatiles from four tomato varieties grown in open-field and high-tunnel systems. Univariate and multivariate analyses were performed to identify the influence of production system on tomato volatiles.

Results and conclusions

The extraction of 2 g tomato samples at 60 °C for 45 min using divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS) fiber gave the maximum amounts of volatiles. This optimized method was used to identify and quantify 41 volatiles from four tomato varieties. The levels of β-damascenone were higher in the high-tunnel tomatoes and geranylacetone was higher in open-field tomatoes. These two volatile compounds could be considered as biomarkers for tomatoes grown in high-tunnel and open-field production systems. This study is the first report comparing volatiles in tomatoes grown in high-tunnel and open-field conditions, and our results confirmed that there is a critical need to adopt biomarker-specific production systems to improve the nutritional and organoleptic properties of tomatoes.

     

Rhythms of volatiles release from healthy and insect-damaged Phaseolus vulgaris

The release rhythm of volatiles is an important physiological characteristic of plants, because the timing of release can affect the function of each particular volatile compound. However, most studies on volatiles release rhythms have been conducted using model plants, rather than crop plants. Here, we analyzed the variations in volatile compounds released from healthy and leafminer (Liriomyza huidobrensis)-infested kidney bean (Phaseolus vulgaris), an important legume crop plant, over a 24 h period. The constituents of the volatiles mixture released from plants were analyzed every 3 h starting from 08:00. The collected volatiles were identified and quantified by gas chromatography–mass spectrometry. Undamaged kidney bean plants released trace amounts of volatiles, with no obvious release rhythms. However, leafminer-damaged plants released large amounts of volatiles, in two main peaks. The main peak of emission was from 17:00 to 20:00, while the secondary peak was in the early morning. The terpene volatiles and (Z)-3-hexenyl acetate showed similar rhythms as that of total volatiles. However, the green leaf volatile (Z)-3-hexen-ol was emitted during the night with peak emission in the early morning. These results give us a clear picture of the volatiles release rhythms of kidney bean plants damaged by leafminer.Keywords : green leaves volatiles, Liriomyza huidobrensis, rhythm, terpene, (Z)-3-hexen-ol     

A method for the solvent extraction of low-boiling-point plant volatiles

A new method has been developed for the extraction of volatiles from plant materials and tested on seedling tissue and mature leaves of Arabidopsis thaliana, pine needles and commercial mixtures of plant volatiles. Volatiles were extracted with n-pentane and then subjected to quick distillation at a moderate temperature. Under these conditions, compounds such as pigments, waxes and non-volatile compounds remained undistilled, while short-chain volatile compounds were distilled into a receiving flask using a high-efficiency condenser. Removal of the n-pentane and concentration of the volatiles in the receiving flask was carried out using a Vigreux column condenser prior to GC-MS. The method is ideal for the rapid extraction of low-boiling-point volatiles from small amounts of plant material, such as is required when conducting metabolic profiling or defining biological properties of volatile components from large numbers of mutant lines.     

Synergism between aromatic compounds and green leaf volatiles derived from the host plant underlies female attraction in the oriental fruit moth

Blends of volatile compounds emitted by host plants are known to mediate the attraction of gravid female herbivores to oviposition sites, but the role of individual odor components is still little understood. We characterized the olfactory response of mated female Cydia (Grapholita) molesta (Busck) (Lepidoptera: Tortricidae) to synthetic mixtures of compounds emitted by peach shoot, a key host plant of this herbivore, and investigated the role of important constituents of bioactive mixtures in moth attraction. Relative ratios of constituents of the mixtures corresponded to the natural ratio of volatile compounds collected in the plant's headspace. A significant attractant effect was found for a comparatively complex 10‐compound mixture that included four green leaf volatiles [(Z)‐3‐hexen‐1‐ol, 1‐hexanol, (E)‐2‐hexenal, and (Z)‐3‐hexen‐1‐yl acetate], five aromatics (benzaldehyde, methyl salicylate, methyl benzoate, benzonitrile, and phenylacetonitrile), and a carboxylic acid (valeric acid). Using a subtraction approach, the number of compounds was progressively decreased, resulting in a bioactive 5‐compound mixture composed of two constituents, green leaf volatiles and aromatic compounds. Further evaluations revealed that benzaldehyde and benzonitrile must be present in association with three distinct green leaf volatiles to produce an attractant effect on the female moths. This 5‐compound mixture was as attractive as natural peach shoot volatiles, which are known to comprise over 20 compounds. Results are discussed in light of the documented synergistic effect between the three general green leaf volatiles and the two specific aromatic compounds.     

Roles of (Z)-3-hexenol in plant-insect interactions

Green leaf C6-volatiles are among the most important herbivore-induced plant volatiles (HIPVs). They play important roles in mediating the behavior of herbivores and their natural enemies, and in triggering the plant-plant communication to prevent further attacks. Recently, wound-induced ubiquitous (Z)-3-hexenol, a C6-alcohol synthesized in the lipoxygenase/HPL pathway, was proved to be the most important info chemical for the herbivore repellence/attraction and natural enemy attraction in tritrophic interactions, as well as for the induction of gene expression in neighboring unattacked plants. In spite of the conflict functions of (Z)-3-hexenol in direct and indirect plant defenses, its positive roles in the indirect defense and the priming effect are consistent. Therefore, this compound can be used to develop novel insect pest control strategies.Key words: green leaf volatiles, (Z)-3-hexenol, direct defense, indirect defense, primingTo date, nearly 2000 volatile compounds have been identified in plant species from over 90 families.¹ These compounds are released from plant organs above or below the ground, and some are induced by biotic activities. Herbivore feeding stimulates the plants to release green leaf volatiles (GLVs), terpenoids, nitrogen-containing nitriles and oximes, methyl salicylate, etc. Production of these volatiles by plants involves at least three biosynthetic pathways: the fatty acid/lipoxygenase pathway for green leaf volatiles, the isoprenoid pathway for terpenoids, and the shikimic acid pathway for methyl salicylate.² Herbivore-damaged plants emit some of the most common GLVs and terpenoids that play important roles in mediating the behaviors of herbivores and their natural enemies, as well as in triggering the plant-plant communication.¹ Recently, functional studies on green leaf C6-volatiles have received wide attention and made exciting progresses. Especially, accumulating evidences on the C6-volatile (Z)-3-hexenol support its role in mediating indirect defense responses of plant.     

Differential electroantennogram response of females and males of two parasitoid species to host-related green leaf volatiles and inducible compounds

Parasitoids employ different types of host-related volatile signals for foraging and host-location. Host-related volatile signals can be plant-based, originate from the herbivore host or produced from an interaction between herbivores and their plant host. In order to investigate potential sex- and species-related differences in the antennal response of parasitoids to different host-related volatiles, we compared the electroantennogram (EAG) responses of both sexes of the specialist parasitoid, Microplitis croceipes (Cresson), and the generalist, Cotesia marginiventris (Cresson), to varying doses of selected plant-based host-related volatiles: two green leaf volatiles (cis-3-hexenol and hexanal) and three inducible compounds (cis-3-hexenyl acetate, linalool, and (E,E)-alpha-farnesene). Mating had no significant effect on EAG response. Females of both species showed significantly greater EAG responses than conspecific males to green leaf volatiles, which are released immediately after initiation of herbivore feeding damage. In contrast, males showed greater responses than conspecific females to inducible compounds released much later after initial damage. Cotesia marginiventris females and males showed greater EAG responses than counterpart M. croceipes to the tested compounds at various doses, suggesting that the generalist parasitoid shows greater antennal sensitivity than the specialist to the tested host-plant volatiles. These results are discussed in relation to the possible roles of green leaf volatiles and inducible compounds in the ecology of female and male parasitoids.     

Differential volatile signatures from skin, naevi and melanoma: a novel approach to detect a pathological process

Background

Early detection of melanoma is of great importance to reduce mortality. Discovering new melanoma biomarkers would improve early detection and diagnosis. Here, we present a novel approach to detect volatile compounds from skin.

Methods and Findings

We used Head Space Solid Phase Micro-Extraction (HS-SPME) and gas chromatography/mass spectrometry (GC/MS) to identify volatile signatures from melanoma, naevi and skin samples. We hypothesized that the metabolic state of tissue alters the profile of volatile compounds. Volatiles released from fresh biopsy tissue of melanoma and benign naevus were compared based on their difference in frequency distribution and their expression level. We also analyzed volatile profiles from frozen tissue, including skin and melanoma.

Conclusions

Three volatiles, 4-methyl decane, dodecane and undecane were preferentially expressed in both fresh and frozen melanoma, indicating that they are candidate biomarkers. Twelve candidate biomarkers evaluated by fuzzy logic analysis of frozen samples distinguished melanoma from skin with 89% sensitivity and 90% specificity. Our results demonstrate proof-of-principle that there is differential expression of volatiles in melanoma. Our volatile metabolomic approach will lead to a better understanding of melanoma and can enable development of new diagnostic and treatment strategies based on altered metabolism.     

Quantitative and qualitative differences in C6‐volatile production from the lipoxygenase pathway in an alcohol dehydrogenase mutant of Arabidopsis thaliana

Six‐carbon (C₆) volatile products are released from the enzymatic action of hydroperoxide lyase (HPL), a component of the lipoxygenase (LOX) pathway and form the basis of the "green‐note" flavour characteristic of many consumed plant products. Arabidopsis leaf tissue contains the C₆‐aldehydes hexanal, and trans ‐2‐hexenal as well as the C₆‐alcohols: hexanol, and 3‐hexenol. Interconversion between C₆‐aldehydes and alcohols is thought to proceed through the action of alcohol dehydrogenase (ADH). Using an ADH mutant of Arabidopsis , we have shown that there are large quantitative and qualitative differences in the accumulation of C₆‐volatiles in the absence of ADH activity. The total quantity of LOX‐derived volatiles is greater on a fresh weight basis in the ADH mutant. Qualitatively, hexanol and 3‐hexenol levels are approximately 62% and 51% lower in the mutant, respectively, whereas levels of hexenal are approximately 10‐fold higher. Hexanal accumulation, however, is unaffected in the mutant. The altered profile of LOX‐derived volatiles does not have an effect on the steady‐state levels of mRNA for allene oxide synthase (AOS) or LOX. HPL activity and mRNA quantity, however, are higher in the mutant relative to wild type, suggesting that altered product levels in the mutant affect HPL regulation.     

Two sesquiterpene synthases are responsible for the complex mixture of sesquiterpenes emitted from Arabidopsis flowers

Despite the fact that Arabidopsis is largely self-pollinating, its flowers emit a complex mixture of terpene volatiles consisting predominantly of a large group of over 20 sesquiterpenes. Here we report that only two terpene synthases, encoded by the florally expressed genes At5g23960 and At5g44630, are responsible for the formation of virtually all sesquiterpenes found in the Arabidopsis floral volatile blend. Two independent mutant lines with T-DNA insertions in the previously identified At5g23960 gene lacked the emission of three sesquiterpenes, including the main sesquiterpene volatile (E)-beta-caryophyllene, confirming the previous in vitro functional assignment for this gene. Flowers of a mutant line carrying a T-DNA insertion in gene At5g44630 emitted these three sesquiterpenes, but did not emit any of the remaining sesquiterpene volatiles. An At5g44630 cDNA was expressed in Escherichia coli and the produced protein catalyzed the conversion of farnesyl diphosphate into over 15 sesquiterpenes in similar proportions to those found in the floral volatile blend. At5g23960 and At5g44630 promoter-beta-glucuronidase (GUS) fusion experiments demonstrated that both genes are expressed in several parts of the Arabidopsis flower, with strong At5g23960 promoter-GUS activity in the stigma and strong expression of At5g44630 in intrafloral nectaries. Given the previously reported antimicrobial activity of terpenes, their production in stigmas and nectaries may serve to inhibit microbial infection at these vulnerable sites. A survey of 37 Arabidopsis thaliana ecotypes revealed quantitative, but almost no qualitative, variations of floral monoterpene and sesquiterpene emissions suggesting that floral terpene volatiles must play some significant role in the life of the Arabidopsis plant.