Optimization and comparison of three methods for extraction of volatile compounds from Cyperus rotundus evaluated by gas chromatography-mass spectrometry

The essential oil of Cyperus rotundus has multiple pharmacological activities. Therefore, the extraction with high yield and quality is very important for preparation of essential oil of C. rotundus. In this paper, three methods, namely hydrodistillation (HD), pressurized liquid extraction (PLE) and supercritical fluid extraction (SFE), for extraction of volatile compounds from C. rotundus were optimized and compared by gas chromatography-mass spectrometry. Among eight identified compounds in C. rotundus, five components including alpha-copaene, cyperene, beta-selinene, beta-cyperone and alpha-cyperone were quantitatively determined or estimated using alpha-cyperone as standard, which showed that PLE had the highest extraction efficiency, while SFE had the best selectivity for extraction of beta-cyperone and alpha-cyperone. The contents of ingredients from C. rotundus extracted with HD, PLE and SFE are significantly different, which suggest that comparison of chemical components and pharmacological activities of different extracts is helpful to elucidate the active components in C. rotundus and control its quality.     

川芎药材超临界CO2萃取与水蒸气蒸馏提取挥发油比较研究

目的 通过对不同工艺下得到的川芎挥发油的比较,阐明提取工艺对其有效成分的影响。方法 分别采用超临界CO₂萃取和水蒸气蒸馏法提取川芎挥发油,再通过气相色谱-质谱(GC-MS)联用技术分析鉴定其中的主要化学成分及相对含量。结果 2种方法所得川芎挥发油样品中共鉴定18种共有成分,水蒸气蒸馏提取物中苯酞类成分约占61%,单萜类成分约占25%,倍半萜类成分约占10%;超临界CO₂萃取物中苯酞类成分约占97%,单萜类成分约占1%,倍半萜类成分约占0.4%。结论 水蒸气蒸馏法对川芎药材中挥发性较强的单萜和倍半萜类保留相对较强,超临界CO₂萃取法对苯酞类有效成分的提取率相对较高,因此在川芎挥发油入药的中药制剂工艺中,应根据制剂的功效充分考虑提取工艺对有效成分的影响。     

不同方法提取的佛手挥发油气相色谱-质谱分析结果比较

目的：比较分析水蒸气蒸馏法与超临界流体萃取法（SFE）提取佛手挥发油的化学成分和相对含量。方法：运用气相色谱-质谱技术（GC-MS），结合计算机检索对其化学成分进行分离和鉴定，用色谱峰面积归一化法计算各组分的相对含量。结果：水蒸气蒸馏法中鉴定出18种化学成分，SFE中鉴定出16种化学成分，二者的主要成分均含有D-柠檬烯，相对含量前者为57．87％，后者为81．18％；水蒸气蒸馏法提出成分中还含有25．12％的（＋）-2-蒈烯，而SFE法未检测到该成分。结论：两种提取方法成分结果有所不同，但都含有主要成分D-柠檬烯，含量以SFE法为高。     

正交试验优选超临界CO_2流体法萃取藿香正气方中挥发油的工艺

目的:优选超临界CO2流体萃取(SFE-CO2)法萃取藿香正气方中挥发油的工艺条件。方法:以萃取压力、萃取温度、解析釜I压力、解析釜I温度为考察因素,以挥发油萃取率为评价指标,采用L9(34)正交试验表进行工艺优选。结果:最佳工艺为萃取压力25MPa、萃取温度30℃、解析釜I压力9MPa、解析釜I温度45℃;此时挥发油的萃取率最高,为2.057%。结论:应用SFE-CO2法萃取藿香正气方中挥发油,萃取率高,稳定性强,操作纯净、安全且能保持生物活性,是具有相当发展潜力的提取分离方法。     

广陈皮的超临界流体萃取和水蒸气蒸馏挥发油的比较分析

目的:比较两种不同提取方法所得的广陈皮挥发油的挥发性化学成分。方法:分别采用超临界CO2流体萃取(SFE)和水蒸气蒸馏(SD)的方法提取广陈皮的挥发油。运用气相色谱-质谱联用法分离和分析两挥发油的挥发性化学成分,主要成分棕榈酸和D-柠檬烯采用标准品的峰面积增大法进一步确认。结果:从广陈皮SFE和SD挥发油中共鉴定出22个成分,占色谱总流出峰面积的97%以上,两者的提取率分别为5.05%和0.46%。SFE挥发油的主要成分为挥发性较弱、相对分子量较大的棕榈酸,SD挥发油的主要成分为挥发性较强、相对分子量较小的D-柠檬烯。此外,SFE和SD挥发油中均有含量不低的2-甲胺基-苯甲酸甲酯。结论:采用两种方法提取广陈皮所得挥发油的挥发性成分不尽相同,且SFE法的提取率高于SD法;两种方法均能将广陈皮的特征性成分2-甲胺基-苯甲酸甲酯提取出来。     

不同方法提取川芎挥发油的GC-MS分析

目的:对川芎饮片不同提取方法所得挥发油的化学成分进行比较分析。方法:采用气相色谱-质谱联用仪对提取物进行比较分析。结果:4种方法提取的挥发油有9个化合物相同。其中水蒸气蒸馏法提取的挥发油样品中组分种类最多,主要是烯萜类成分。结论:4种提取方法均能提取出川芎挥发油中的主要成分,主要成分为内酯类,相对含量最高的为反式-藁本内酯。     

川芎的适宜采收期和加工方法

目的研究不同采收期和不同加工方法对川芎有效成分含量的影响。方法按《中国药典》2000年版方法分别测定了不同产地、3个采收期和6种干燥加工方法川芎中水分、挥发油的含量,HPLC法测定了阿魏酸的含量,酸性染料比色法测定了总生物碱的含量。结果5月10日~30日,挥发油的含量随采收期的延迟而降低,阿魏酸含量则明显增加;总生物碱含量显著增加。挥发油的含量以水洗后晒干法损失最大,其他方法差异不明显;阿魏酸以微波干燥的含量最高,远红外干燥法的次之;总生物碱以远红外和农户烘干法的含量最高。结论川芎最适宜的采收期以每年5月20日(约农历小满)后10 d为宜;加工方法以产地农户烘干法为宜。     

Identification and comparison of metabolites after oral administration of essential oil of Ligusticum chuanxiong or its major constituent ligustilide in rats

The rhizome of Ligusticum chuanxiong Hort. is a well-known traditional Chinese medicinal herb that is widely used to treat various vascular diseases. The aims of this study were to identify and compare the metabolites in rats after the oral administration of the essential oil of Ligusticum chuanxiong Hort. or its major constituent ligustilide monomer, and to explore the differences in the metabolism of the main active constituent monomer and the active constituent group. High-performance liquid chromatography with diode array detection was used to analyze the metabolites in the urine, feces, and bile of rats dosed with the essential oil or ligustilide. We observed clear differences in the metabolism of the essential oil and ligustilide. The other chemical constituents in the essential oil influenced the metabolism of the main active constituent monomer. Four metabolites in urine were separated, purified, and identified as 6,7-dihydro-6,7-dihydroxyligustilide (M1), 3,5-dihydroxy-3-butylphthalide (M2), 6,7-dihydro-6-hydroxy-7-sulfmethylligustilide (M3), and 5-hydroxy-3-butylidenephthalide (M4) by FT-ICR-MS and 1H-NMR. M2 and M3 are novel metabolites. Based on the profiles of these metabolites, the pathway of ligustilide metabolism in rats is proposed.     

超临界CO2提取的百合挥发油化学成分的GC-MS分析

目的 研究百合挥发油化学成分。方法 采用超临界CO₂提取方法,GC-MS联用技术鉴定百合挥发油成分,并用峰面积归一化法测定各成分的相对百分含量。结果 超临界CO₂流体萃取法提取的挥发油共鉴定出50种成分,占挥发油总成分的72.96%。结论 超临界提取百合挥发油成分多、效率高,与水蒸气蒸馏提取出的挥发油成分有差异。     

GC—MS法对中药复方酸枣仁汤及其主要单味药超临界流体萃取物的成分分析

目的:研究中药复方酸枣仁汤及其主要单味药超临界流体萃取物化学成分的差异。方法:采用超临界流体萃取技术和传统水蒸气蒸馏法制备供试品溶液,运用 GC-MS 法对复方及其主要单味药的超临界流体萃取物和水蒸气蒸馏物进行化学成分分析。结果:共鉴定出37个化合物,其中2,4-癸二烯醛、棕榈酸甲酯、棕榈酸乙酯为首次从酸枣仁油中发现;4-甲基-3-环戊烯-1醇、安息油、4-羟基-3-甲氧基安息香醛、1,2,3,4-四氢萘烯、4-甲基苯酚、3-甲基-7-甲氧基苯并吡喃、4-甲氧基苊烯、4,5-二甲氧基苯环丁烯醇8个化合物为首次从川芎油中发现。结论:复方酸枣仁汤与其主要单味药超临界流体萃取物的化学成分存在一定差异,研究结果为探讨复方共提和单味药分提的化学成分变化规律提供了一定的依据。     

超临界流体萃取法测定川芎中藁本内酯含量的研究

系统研究了压力、温度、静态萃取时间、动态萃取量、改性剂加入量等因素对超临界流体萃取(SFE)中药川芎中藁本内酯的影响,确定最佳萃取条件为:压力27.6MPa,温度40℃,静态萃取时间3min,动态萃取量7ml,改性剂加入量0.1ml。并对超临界流体萃取的收集方法作了研究,发现固液收集法在收集效率和精密度方面比溶剂收集法效果好。并用离线的SFE RPHPLC对川芎药材中藁本内酯的含量进行了测定。     

川芎挥发油的GC-MS指纹图谱研究

目的:建立川芎挥发油的气相色谱-质谱(GC-MS)指纹图谱。方法:采用超临界CO2流体萃取法提取川芎挥发油,用GC-MS联用技术对其进行指纹图谱研究,色谱柱为SGE-30QC3/AC225弹性石英毛细管柱(30m×0.32mm×0.25μm),进样口温度为280℃,程序升温,载气为氦气,进样量为1μL,分流比为10:1;采用电子轰击离子源(EI),离子源温度为200℃,接口温度为250℃,检测电压为0.8kV,质量扫描范围为30～400amu;并采用"中药指纹图谱计算机辅助相似度评价软件"计算相似度。结果:本方法有较好的重复性、精密度和稳定性(RSD均<3%);川芎挥发油主要含有α-蒎烯、胡萝卜烯醇、川芎内酯、藁本内酯、棕榈酸,这5种成分构成了川芎挥发油GC-MS特征指纹图谱;10批川芎挥发油的相似度均>0.92。结论:本研究可为川芎的质量控制提供依据。     

HS-SPME-GC-MS联用对当归各药用部位挥发油成分的分析

目的：分析比较当归各药用部位的挥发油成分。方法：通过单因素考察确定最佳萃取条件,采用顶空固相微萃取法（HS-SPME）与气相色谱-质谱（GC-MS）联用法测定当归各药用部分挥发性化学成分,通过面积归一法确定各成分含量。结果：最终确定条件为称量量0.5 g,萃取温度90℃,萃取时间20 min,解吸附时间3 min。全当归药材粉末出峰94个,当归头药材粉末出峰64个,当归身药材粉末出峰62个,当归尾药材粉末出峰73个,全当归、当归头、当归身和当归尾定性成分含量分别为99.994%,99.747%,97.754%和99.988%。结论：当归挥发性成分主要为藁本内酯且藁本内酯含量：当归身>当归尾>当归头>全当归。     

三种方法提取防风挥发油的气质联用成分分析

目的对3种不同方法提取的防风挥发油化学成分进行分析。方法采用水蒸气蒸馏法（SD）、溶剂提取法（SX）、超临界CO2流体萃取法（SFE-CO2）提取防风挥发油,用气相-质谱（GC-MS）法对化学成分进行鉴定。结果水蒸气蒸馏法提取挥发油共鉴定了13种化学成分;溶剂提取法提取的挥发油共鉴定了18种化学成分;超临界CO2流体萃取法提取的挥发油共鉴定了21种化学成分。结论超临界CO2流体萃取法提取的挥发油更全面地反映药材中的化学成分。     

超临界CO_2萃取当归挥发油的研究及藁本内酯的HPLC法测定

目的采用超临界CO2萃取当归挥发油,并对其中主要成分藁本内酯进行含量测定,以确定当归的最佳萃取工艺条件。方法运用正交设计法,通过对影响萃取的主要因素:萃取压力、萃取温度和萃取时间进行考察,以萃取物得率与高效液相色谱法测定的藁本内酯含量作为指标综合评分,筛选出最佳萃取工艺条件。结果当归挥发油超临界CO2萃取的最佳工艺为:萃取压力35MPa,萃取温度35℃,萃取时间2h,该条件下萃取率可达4.06%。经高效液相色谱分析,藁本内酯含量为19.59%。结论SFE-HPLC方法简单可靠,可用于当归藁本内酯的提取及分析。     

超临界CO_2流体萃取榧树假种皮挥发油化学成分的研究

目的:研究超临界CO_2流体萃取榧树假种皮挥发油并分析挥发油组成。方法:采用超临界CO_2流体萃取技术提取其精油部分,经二级解析分离出精油Ⅰ和Ⅱ,并用气相色谱-质谱-计算机联用技术进行分离和鉴定。结果:挥发油的最佳萃取条件为压力32mPa、温度40℃、时间1h。榧树假种皮挥发油含量为5.83％,挥发油由柠檬烯等48个化学成分组成。结论:超临界CO_2流体萃取榧树假种皮挥发油具有提取时间短、提取完全等优点。     

正交试验优选左金漂浮片中挥发油提取工艺研究

目的优选左金漂浮片中吴茱萸挥发油的最佳提取工艺。方法采用正交试验，以挥发油得率为指标，比较超临界萃取及水蒸气蒸馏两种提取方法的提取效果。结果吴茱萸挥发油水蒸气蒸馏的最佳提取工艺为A3B1C3，超临界萃取的最佳提取工艺为A3B1C3，后者明显优于前者。结论与水蒸气蒸馏法相比，超临界萃取法具有耗时少、效率高和对提取后物料破坏较小等优点，适合于批量生产，便于制剂的后续提取工艺制备。     

Phytochemistry,biological properties and quality control of Chuanxiong Rhizoma: a review

Chuanxiong Rhizoma is a widely used Chinese herbal medicine in the past 2000 years. Chuanxiong Rhizoma is composed of volatile oils, phthalide lactones, phenolic acids, polysaccharides and other compounds. To date, more than 149 compounds in Chuanxiong Rhizoma have been isolated and identified, and some of them have been reported to possess promising biological properties on cardiovascular and central nervous system disorders besides their anti-cancer and antioxidant effects. Modulation of inflammatory mediators and apoptotic factors are believed to contribute to its bioactivities. Analytical methods, such as HPLC, GC and UPLC, are employed for qualitative evaluation of Chuanxiong Rhizoma. In this work, harvest period, growing habitat, processing method and storage, which can affect the quality of Chuanxiong Rhizoma, were also discussed. Comprehensive quality control methods should be developed to ensure the safety, quality and efficacy use of Chuanxiong Rhizoma. Herein, we collected and analyzed the literature of Chuanxiong Rhizoma published on CNKI, ScienceDirect, Springer link, Wiley and PubMed in past two decades, and up-to-date information of Chuanxiong Rhizoma was provided in this paper. We suggested ligustilide, butylidenephthalide and total senkyunolides as the chemical markers to evaluate the quality of Chuanxiong Rhizoma. Additionally, the influences of soil conditions and processing methods on Chuanxiong Rhizoma as future research perspectives should also be further assessed.     

黑种草籽挥发油化学成分分析

目的提取、分析黑种草籽的挥发性成分。方法用超临界二氧化碳萃取法提取、气相色谱-质谱分析（GC-MS）方法测定挥发油的成分,质谱图用NIST图谱库检索,对其挥发性成分进行定性、定量。结果得到了29个定性结果,鉴定出29个化合物,主要化合物有异长叶烯（isolongifolene）38.01%,松烯（sabinene）11.23%,4-（1-异丙基）-苯甲醛（4-methylethyl-benzaldehyde）7.67%,5-异丙基-2-甲基-1,4苯醌（5-isopropyl-2-methyl-1,4-benzoquin）7.02%,乙酸龙脑酯（bomylacetate）5.27%,樟脑（camphor）4.98%,冰片（bornol）3.13%,左旋薄荷酮（L-menthone）2.25%。结论用超临界二氧化碳萃取法得到的挥发油的纯度高。     

Influence of extraction methods on the composition of essential oils

The aim of this work is to demonstrate our results on comparison of composition of essential oil fractions obtained by traditional steam distillation and supercritical fluid extraction. The plant materials for the various extraction methods were selected from the Lamiaceae, Apiaceae and Asteraceae families. For the supercritical fluid extraction (SFE) carbon dioxide was used as supercritical solvent. The extracts were collected by stage wise precipitation in two separators. The waxy product and extract rich in essential oil were collected in the 1st and in the 2nd separator respectively. The traditional water steam distillation (SD) was carried out in the special apparatus of the Hungarian Pharmacopoea (7th ed.). GC analysis was carried out on capillary silica fused columns coated with DB-1701 and the specific chiral columns coated with Rt-beta DEX m or Rt-beta DEX sm. Comparing the composition of steam distilled oils with that of volatile SFE fractions the following general characteristics were established. The SFE fractions were richer in monoterpene-esters and poorer in alcohols than the traditional essential oils (clary sage, lavander, moldavian dragonhead). Regarding the distribution of the monoterpene and sesquiterpene compounds, the SFE fractions contained sesquiterpenes in higher percentage than the distilled oils (Salvia fruticosa). Furthermore, the proportion of sesquiterpenes increased in SFE fractions collected successively with time (Salvia officinalis) similar to the ratio of oxygenated monoterpenes to monoterpene hydrocarbons (Rosmarinus officinalis). The phtalides of lovage (Satureja hortensis) did not show regular change during the supercritical extraction. In other cases it was verified that part of the mono- and sesquiterpenes were present originally in bound form (glycosides) in plants. Thus they appeared only in essential oil fractions after previous acidic treatment (Thymus, Origanum, Satureja species). During the super-critical extraction the azulenogene sesquiterpene lactones did not transform to azulenes (chamomile, yarrow), but SFE fractions of some Asteraceae plants contained sesquiterpene-gamma-lactones of unchanged structure.