固相微萃取技术及其在药物分析中的应用

本文评述了固相微萃取技术的工作原理、萃取装置、操作过程及技术条件,介绍了固相微萃取技术在药物分析相关领域的应用近况,展望了这一技术的发展前景。     

固相微萃取技术在白蚁表皮碳氢化合物分析中的应用

利用固相微萃取(SPME)技术对白蚁表皮碳氢化合物进行萃取,选出较适合的萃取头,适宜的萃取温度和时间,同时利用GC-MS分析白蚁表皮碳氢化合物的组成成分及含量。结果表明,该白蚁表皮中含有24种碳氢化合物,主要为11,13-二甲基二十六烷烃(33.34%)和2,4,6,-三甲基二十七烷烃(15.75%)。     

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.     

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。     

午子绿茶香气物质固相微萃取GC-MS分析

选用春、秋两季的午子绿茶样品,采用固相微萃取法提取、富集其芳香物质,经GC-MS分析,共鉴定出112种化合物,其中春茶102个组分、秋茶96个组分.不同生产季节绿茶中香气物质构成种类基本相同,其含量分布存在差异.苯乙醇、苯甲醇、2,6-二叔丁基对甲苯酚、壬醛、咖啡因、香叶醇、β-芳樟醇、1,2,4a,5,8,8a-六氢-4,7-二甲基-1-异丙基萘等化合物作为午子绿茶中主要的香气物质,对于构成其特征风味具有重要作用.     

收集挥发性样品的吸附剂法和固相微萃取法

本文介绍挥发性信息化学物质的两种收集方法——吸附剂收集法和固相微萃取收集法,以植物挥发性化学物质收集为例为例,详述了两种方法的原理、步骤、注意事项和实例,最后比较了两种收集方法的优缺点,供相关工作者使用时参考。     

固相微萃取-气质法测定土壤挥发性抑菌物质

运用固相微萃取．气相色谱,质谱法(SPME-GC／MS),测定了参与土壤抑真菌作用的土壤挥发性成分和土壤细菌挥发性代谢物。通过比较土壤来源和土壤细菌来源的挥发性抑菌成分,发现在强挥发性抑菌土壤和土壤细菌代谢物中普遍存在着三甲胺、二甲基二硫醚、3-甲基-2-戊酮、甲基吡嗪、2,5-二甲基吡嗪、N,N-二甲基辛胺、十九烷等化合物。这些化合物很有可能就是参与土壤抑菌作用,特别是挥发性物质抑菌作用的主要成分。另外,为深入了解土壤中参与抑菌作用的挥发性化合物提供了简便有效的方法。     

落叶松（Larix gemelinii）苗挥发物两种收集方法的对比分析

采用预浓缩和固相微萃取(SPME)2种方法对2年生兴安落叶松苗挥发物成分进行采集并做GC-MS分析.结果表明,落叶松苗挥发物主要成分为萜烯类化合物,其中α-蒎烯、4(10)-侧柏烯、莰烯、β-蒎烯、D-柠檬烯和β-水芹烯所占比例较高;整株、针叶和树皮挥发物的组成和含量存在差异,针叶中饱和烷烃含量较高,树皮中倍半萜烯、醇、酮和酯类挥发物含量较高,并且这2种组织经SPME采集检测的挥发物中石竹烯和α-蛇麻烯含量增加幅度较大,它们可能在昆虫近距离寄主定位中起作用.对2种气体收集方法的分析比较,认为预浓缩结果接近落叶松挥发物组分和含量的自然特征,SPME受萃取头纤维特性的影响,对倍半萜稀类化合物吸附作用强,对烷烃吸附差,并且不同化合物的同时存在可能影响其吸附效率,导致检测量准确度下降,因此预浓缩法更适合植物挥发物定性和定量分析.     

顶空固相微萃取/气相色谱/质谱法分析河南产紫丁香挥发性成分

采用顶空固相微萃取和气质联用技术（HS—SPME—GC—MS）,首次分析了河南产紫丁香花营和花的挥发性成分。从紫丁香花蕾和花中共鉴定了65种组分,其中17个成分是二者共有的;酯类化合物只存在于花蕾中,而酸类和醇类化合物在花中的含量较高。这表明酯类化合物在紫丁香花开放的过程中可能逐步被植物体内的水解酶水解成酸类和醇类化合物。     

固相微萃取—气相色谱—质谱联用分析蓼实挥发性成分

察了萃取样品温度、萃取纤维吸附时间等因素对于固相微萃取蓼实挥发性成分的影响,确定较佳的实验条件为：萃取样品温度60℃,萃取纤维吸附时间60 min,脱附温度250℃,脱附时间5 min。用气相色谱—质谱联用技术测定上述条件所得蓼实挥发性化学成分,并鉴定出其中43种,占总峰面积的76.73%。其中含量较高的物质有：罗汉柏烯 (6.99%),丁香烯 (5.59%),2,5,5,8 a-四甲基-6,7,8,8a 四氢-5H-萘-1-酮(5.52%),α-丁香烯(4.29%),1,2,4a,5,6,8a-六氢-4,7-二甲基-1-异丙基萘(4.04%),环氧石竹烯(3.60%),α-香附酮(3.54%),4,5,5a,6,6a,6b-6氢-4,4,6b-三甲基-2-乙烯基-2H-环丙香豆酮(3.54%),香叶基丙酮(3.48%)。     

豇豆与菜豆挥发物中美洲斑潜蝇引诱成分的分析与鉴定

应用固相微萃取技术(SPME)和自行设计的挥发物收集装置以及GC-MS系统,对美洲斑潜蝇主要寄主植物豇豆与菜豆的挥发物进行收集和分析.鉴定出两种寄主植物的挥发物主要成分均为2-己烯醛、3-己烯醇、2-己烯醇、1-辛烯醇、3-己烯醇醋酸酯、α-紫罗酮、β-紫罗酮.通过与标准品的对照,测定了7种主要挥发性化合物之间的相对比例,并据此配制成人工引诱剂,进行了美洲斑潜蝇成虫的田间诱捕试验.结果证实所配制的引诱剂对成虫具有一定的引诱效果,其中α-紫罗酮和β-紫罗酮很可能在美洲斑潜蝇成虫寻找寄主植物的定向机制中起着重要作用.     

嘉陵江小三峡硅藻群落结构及水环境

为了研究嘉陵江小三峡硅藻群落结构及水质状况,按枯水期（2006年11月—2007年3月）、平水期（2007年4—6月）和丰水期（2007年7—9月）分别采集6个样点的硅藻样品,分析其物种组成、细胞密度、多样性和均匀度指数及硅藻商等,并结合理化指标对水质进行评价。结果表明：嘉陵江小三峡共有硅藻78种（含变种和变型）,隶属2纲、10科、20属;硅藻细胞密度为0.53×10⁴～3.58×10⁴ ind·L^-1,总平均1.55×10⁴ ind·L^-1;多样性指数值（H′）为1.62～2.96,总平均2.33;均匀度指数值（E）为0.28～0.56,总平均0.42;嘉陵江小三峡水质总体上属β-中污水体,沙溪、北温泉水质较好,属微污水体,碚石、牛石沱水质次之,为β-中污水体,盐井、毛背沱水质最差,为α-中污水体。     

灰树花子实体与发酵菌丝体挥发性化合物分析

分别采用固体栽培法和液体发酵法对同一灰树花菌株进行培养,获得灰树花子实体和发酵菌丝体,并采用顶空固相微萃取(HS-SPME)和气相色谱-质谱(GC-MS)分析技术对子实体和菌丝体的挥发性化合物进行了分析比较。结果表明,灰树花子实体和发酵菌丝体分别含有62种和94种挥发性物质,其中37种为相同物质,分别占总挥发性物质的86.81%和84.28%。灰树花子实体中含有醇(14种)、酮(13种)、醛(12种)、酸(7种)、酯(5种)等物质,主要以酯类(42.80%)、醛类(35.14%)物质为主,其中乙酸乙酯的相对含量达到了42.57%。发酵菌丝体中主要含有醛(22种)、酮(17种)、醇(16种)、酯(13种)、酸(12种)等挥发性物质,其中醛类、酯类、酸类、醇类分别占47.0%、10.90%、7.48%和5.94%。异戊醛和地衣酚的含量分别达到了23.31%、15.41%。     

不同产地丁香挥发性成分分析

采用顶空固相微萃取-气相色谱-质谱联用技术对产自国内外4个地区的丁香(Eugenia caryophyllata Thunb.)挥发性成分进行了检测,并以峰面积归一化法计算了各组分的相对含量。结果显示,从4个产地13批次丁香中共鉴定出挥发性成分72种(匹配度均高于75),其中,主要成分含量排名前两位的均为丁香酚、(-)-α-芹子烯,排名第三位的挥发性成分分别为:顺式-α-没药烯(马达加斯加塔马塔夫,平均为3.75%)、罗勒烯(中国广东,平均为4.21%)、亚麻三烯(中国广西,平均为3.74%)、丁香烯(印度尼西亚爪哇岛,平均为3.60%)。表明同一产地的丁香挥发性成分具有一定的相似性,不同产地丁香挥发性成分也有一定的差异,这对丁香产地的鉴别具有重要意义。主成分分析和聚类分析的结果既可以将4个产地的丁香很好地区分开来,又能反映出它们之间的亲缘关系。本研究采用HS-SPME-GC-MS方法检测不同产地丁香挥发性成分并结合主成分分析(PCA)及聚类分析法,能有效区分4个产地的丁香,该方法可作为丁香产地的鉴别方法,也为进一步比较不同产地丁香挥发性成分的差异及质量控制提供了新思路。     

Determination of dichloromethane, trichloroethylene and perchloroethylene in urine samples by headspace solid phase microextraction gas chromatography-mass spectrometry

A method for the determination of volatile chlorinated hydrocarbons, namely dichloromethane (DCM), trichloroethylene (TCE), and perchloroethylene (PCE), in urine samples was developed using headspace solid phase microextraction (HS-SPME) gas chromatography-mass spectrometry (GC-MS). HS-SPME was performed using a 75 microm Carboxen-polydimethylsiloxane fiber. Factors, which affect the HS-SPME process, such as adsorption and desorption times, stirring, salting-out effect, and temperature of sampling have been evaluated and optimized. The highest extraction efficiency was obtained when sampling was performed at room temperature (22 degrees C), from samples saturated with salt and under agitation. Linearity of the HS-SPME-GC-MS method was established over four orders of magnitude and the limit of detection was 0.005 microg/l for all the compounds. Precision, calculated as %R.S.D. at three different concentration levels, was within 1-8% for all intra- and inter-day determinations. The method was applied to the quantitative determination of TCE and PCE in human urine samples from exposed (TCE, n=5; median, 9.32 microg/l and PCE, n=39; median, 0.58 microg/l) and non-exposed individuals (n=120; median concentrations, 0.64, 0.22 and 0.11 microg/l for DCM, TCE and PCE, respectively. In addition, two cases of acute accidental exposure to DCM are reported, and the elimination kinetics in blood and urine was followed up. The calculated half-lives of urinary and blood DCM were, respectively, 7.5 and 8.1 h for one subject and 3.8 and 4.3 h for the other.     

Identification and profiling of volatile metabolites of the biocontrol fungus Trichoderma atroviride by HS-SPME-GC-MS

In the present study we describe a method, which is based on solid phase microextraction (SPME) coupled to gas chromatography-mass spectrometry (GC-MS) and which can be used for the profiling of microbial volatile organic compounds (MVOCs) in the headspace (HS) of cultures of filamentous fungi. The method comprises the following successive steps: 1. growth of the fungus on a solid culture medium directly in headspace vials, 2. measurement of volatiles by HS-SPME-GC-MS, 3. deconvolution of mass spectra, 4. identification of volatiles by comparison of measured, deconvoluted mass spectra and linear temperature programmed retention indices (LTPRI) on two stationary GC phases with database entries and LTPRI published in the literature, and 5. profiling of the identified MVOCs.The developed method was successfully applied to cultures of the biocontrol fungus Trichoderma atroviride. An in-house library consisting of mass spectra and LTPRI values of fungal VOCs was established and used to study the profiles of MVOCs of this fungus. In total, 25 different MVOCs were identified by applying strict criteria (spectral match factor at least 90% and a maximum relative deviation of LTPRI of ± 2% from literature values). The MVOCs were assigned to the compound classes of alcohols, ketones, alkanes, furanes, pyrones (mainly the bioactive 6-pentyl-alpha-pyrone), mono- and sesquiterpenes, 13 of which have never been reported to be produced by Trichoderma spp. before. Eleven of these volatiles have been additionally confirmed using authentic standards. Finally, time course experiments and cultivation of T. atroviride in the presence of the mycotoxin fusaric acid demonstrated the potential of the method to study the dynamics of MVOC profiles as well as the effect of different environmental/biological conditions on the expression of MVOCs of filamentous fungi.     

Discriminant analysis of volatile organic compounds of Fusarium oxysporum f. sp. cepae and Fusarium proliferatum isolates from onions as indicators of fungal growth

Basal rot is a common onion disease and is mainly caused by Fusarium oxysporum f. sp. cepae and Fusarium proliferatum. To study the possibility of using volatile organic compounds (VOCs) as biomarkers for these fungi, pathogenic isolates of F. oxysporum and F. proliferatum from onions were cultivated in onion medium and VOCs were measured by solid phase microextraction (SPME). Forty-two compounds were detected, and thirty of these compounds were highly related to fungal metabolic activity. Allyl mercaptan was specific to F. oxysporum isolate Fox006. Analysis of the VOCs showed significant differences between the two species and among different isolates within the same species. Sixteen of the VOCs showed were highly positively correlated with the fungal biomass estimated by real-time polymerase chain reaction (PCR). Ethanol, ethyl formate, ethyl acetate, 2-methyl-1-propanol, methyl thioacetate, n-propyl acetate and 3-methyl-1-butanol are volatile metabolites that were potential indicators of Fusarium growth on onions.     

Seasonal, populational and ontogenic variation in the volatile oil content and composition of individuals of Origanum vulgare subsp. Hirtum, assessed by GC headspace analysis and by SPME sampling of individual oil glands

Small-scale GC headspace analyses combined with SPME sampling of individual oil glands have been used to measure the variation in volatile content and composition in and within different oregano plants as affected by age, season and developmental state. The main monoterpenes found were p-cymene, carvacrol and their precursor gamma-terpinene. The early season preponderance of p-cymene over carvacrol was reversed as the season progressed and this pattern could also be seen at any time within the plant, from the latest leaves to be produced (low in cymene) to the earliest (high in cymene). Seedlings from the same mother plant developed this pattern at different rates. Within individual leaves the pattern was not observed, even within the youngest developing leaves. However it was found that the oil composition of individual glands within a single leaf varied considerably, most notably in respect of the production of carvacrol and its isomer thymol.     

顶空固相微萃取-气相色谱质谱法分析山荆子花和叶中的挥发性成分

首次采用顶空固相微萃取-气相色谱质谱法(HS-SPME-GC-MS)分析了盛开期的山荆子花和叶中的挥发性成分,采用保留指数与质谱图库检索的方法定性,利用面积归一化法定量组份含量.鉴定出山荆子花中36种化学成分,占总含量的96.20％,主要为烷烃类(27.00％)、酯类(20.86％)、芳香醇类(11.75％)、萜类(18.78％)化合物,主要成分是2,6-二叔丁基对甲酚(BHT)为14.59％,其次为正十九烷(11.58％)、正二十一烷(8.37％)、法呢烯(8.24％)、罗勒烯(7.77％).鉴定出山荆子叶中22种化学成分,占总含量的98.04％,主要为萜类(50.84％)、烯烃类(25.33％)、酯类(9.73％)化合物,主要成分是法呢烯(40.62％),其次是唯一的烯烃4,8-二甲基-1,3,7-壬烯(26.30％)、罗勒烯(7.27％).     

Analysis of volatile constituents isolated by hydrodistillation and headspace solid-phase microextraction from Adenostyles briquetii Gamisans

The volatile fraction of the whole plant and separated organs of Adenostyles briquetii Gamisans (syn. Cacalia briquetii; family Asteraceae), an endemic species from Corsica, has been studied by headspace solid-phase microextraction (HS-SPME), GC and GC-MS(EI and CI). A total of 141 components were identified, representing 93% of the entire amount. The volatile fraction was characterised by sesquiterpene hydrocarbons (52.8%) and oxygenated sesquiterpenes (25.9%). The major components were germacrene D (18.5%), zingiberene (12.9%) and beta-oplopenone (10.8%). The influence of HS-SPME parameters on the extraction of family components is reported for the first time.