Qualitative and quantitative determination of Atractylodes rhizome using ultra‐performance liquid chromatography coupled with linear ion trap–Orbitrap mass spectrometry with data‐dependent processing

A quick and effective workflow based on ultra‐performance liquid chromatography coupled with electron spray ionization and LTQ‐Orbitrap mass spectrometry (UPLC‐LTQ‐Orbitrap MS) was established for compositional analysis and screening of the characteristic compounds of three species of Atractylodes rhizome for quality evaluation. This technique was employed to determine the seven main components in Atractylodes rhizome samples. Ultimately, 78 constituents were identified; of these, seven characteristic compounds were selected for species discrimination, comprising atractylodin (63), atractylenolide I (43), atractylenolide II (49), atractylenolide III (53), atractylon (69), methyl‐atractylenolide II (54) and (4E,6E,12E)‐tetradecadecatriene‐8,10‐diyne‐1,3‐diacetate (59). The seven main compounds, including six characteristic compounds, were simultaneously determined in 29 batches of Atractylodes rhizome samples. Thus, the method validation showed acceptable results. Quantitative analysis showed significantly different contents of the seven main components among the three species of Atractylodes rhizome, which indicates possible distinctions in the pharmacological effects. This established method can simultaneously provide qualitative and quantitative results for compositional characterization of Atractylodes rhizomes and for quality control.     

基于元素指纹的白术产地溯源及其与土壤的相关性研究

本研究主要采用电感耦合等离子体质谱仪(ICP-MS)等手段测定5个产地白术41种矿质元素含量和土壤41种化学成分指标, 结合聚类分析法、偏最小二乘判别分析(PLS-DA)法和Pearson相关性分析研究了不同产区白术矿质元素特征以及不同产区土壤因子的差异, 并对它们的相关性进行了探讨. 结果表明, 采用无监督的聚类分析和有监督的PLS-DA均能正确判别不同产地的白术和土壤, 其中稀土元素在浙江白术与土壤间呈正相关. 另外, Li、V、Mn、Co、Cu、Rb、Cd、Cs、Ba等微量元素是白术产地溯源的关键因素. 研究发现在白术生产实践中可以通过适量控制种植地的土壤湿度、增施特定肥料等措施调控白术矿质元素含量. 本研究将为解释道地白术的品质形成机制, 保证白术药效质量的有效性提供一定帮助.     

顶空液液萃取-气相色谱-质谱法用于白术挥发性成分的分析

采用高温顶空液相萃取再转移的方法,对中药白术中挥发性成分进行萃取分离富集,采用顶空液液萃取/气相色谱-质谱(HS-LP-LPE/GC-MS)联用法进行测定,并与传统水蒸汽蒸馏法(SD)提取的挥发性成分进行对比。对各种测定条件和影响因素进行了考察,最佳萃取条件为:1.0 mL PEG 400为高温萃取剂,样品用量1.2 g,萃取温度120℃,萃取时间60 min,再将萃取剂用1.0 mL正己烷进行反萃取后进行GC-MS分析;采用HS-LP-LPE/GC-MS鉴定了33个组分,占总组分含量93.18%;SD鉴定了31个组分,占总组分含量97.12%。两种方法共同检测到的组分有29个,均以苍术酮(Atractylone)含量最高。结果表明,两种方法所提取的组分基本相同,可用于白术挥发性成分的测定。     

北苍术和茅苍术挥发油成分的比较

采用气相色谱-质谱联用技术分析了北苍术和茅苍术的挥发油成分,分别鉴定出47和50种化学成分,并测定了其相对含量。 采用色谱指纹图谱八强峰法和分区法,根据色谱峰的保留时间把总离子流色谱图分为5个区,比较了北苍术和茅苍术的挥发油成分。 结果表明,在总离子流色谱图Ⅳ和Ⅴ区中,从北苍术和茅苍术中均检出苍术的主要药效成分β-桉叶油醇、苍术酮和苍术素,且相对含量较高。 这表明《中国药典》将北苍术、茅苍术统称为苍术具有一定的科学性。     

Quantitative and fingerprinting analysis of Atractylodes rhizome based on gas chromatography with flame ionization detection combined with chemometrics

This study assessed the feasibility of gas chromatography with flame ionization detection fingerprinting combined with chemometrics for quality analysis of Atractylodes rhizome. We extracted essential oils from 20 Atractylodes lancea and Atractylodes koreana samples by hydrodistillation. The variation in extraction yields (1.33–4.06%) suggested that contents of the essential oils differed between species. The volatile components (atractylon, atractydin, and atractylenolide I, II, and III) were quantified by gas chromatography with flame ionization detection and confirmed by gas chromatography with mass spectrometry, and the results demonstrated that the number and content of volatile components differed between A. lancea and A. koreana. We then calculated the relative peak areas of common components and similarities of samples by comparing the chromatograms of A. lancea and A. koreana extracts. Also, we employed several chemometric techniques, including similarity analysis, hierarchical clustering analysis, principal component analysis, and partial least‐squares discriminate analysis, to analyze the samples. Results were consistent across analytical methods and showed that samples could be separated according to species. Five volatile components in the essential oils were quantified to further validate the results of the multivariate statistical analysis. The method is simple, stable, accurate, and reproducible. Our results provide a foundation for quality control analysis of A. lancea and A. koreana.     

Differentiation of essential oils in Atractylodes lancea and Atractylodes koreana by gas chromatography with mass spectrometry

Atractylodes rhizome is a valuable traditional Chinese medicinal herb that comprises complex several species whose essential oils are the primary pharmacologically active component. Essential oils of Atractylodes lancea and Atractylodes koreana were extracted by hydrodistillation, and the yield was determined. The average yield of essential oil obtained from A. lancea (2.91%) was higher than that from A. koreana (2.42%). The volatile components of the essential oils were then identified by a gas chromatography with mass spectrometry method that demonstrated good precision. The method showed clear differences in the numbers and contents of volatile components between the two species. 41 and 45 volatile components were identified in A. lancea and A. koreana, respectively. Atractylon (48.68%) was the primary volatile component in A. lancea, while eudesma‐4(14)‐en‐11‐ol (11.81%) was major in A. koreana. However, the most significant difference between A. lancea and A. koreana was the major component of atractylon and atractydin. Principal component analysis was utilized to reveal the correlation between volatile components and species, and the analysis was used to successfully discriminate between A. lancea and A. koreana samples. These results suggest that different species of Atractylodes rhizome may yield essential oils that differ significantly in content and composition.     

Simultaneous determination of atractylenolide II and atractylenolide III by liquid chromatography-tandem mass spectrometry in rat plasma and its application in a pharmacokinetic study after oral administration of Atractylodes Macrocephala Rhizoma extract

Atractylenolide II (AII) and atractylenolide III (AIII) are the major active components in Atractylodes Macrocephala Rhizoma (AMR). In this study, a sensitive, rapid and selective liquid chromatography–tandem mass spectrometry (LC‐MS/MS) method was developed and validated for the simultaneous determination of AII and AIII in rat plasma using loliolide as internal standard (IS). After protein precipitation with ethyl acetate, the analytes were injected into an LC‐MS/MS system for quantification. Chromatography was performed using a C₁₈ column, eluting with water and acetonitrile (45:55, v/v) at 0.2 mL/min. All analytes including IS were monitored under positive ionization conditions by multiple reaction monitoring with an electrospray ionization source. The validated method was successfully applied to the pharmacokinetic study of AII and AIII in rat plasma after oral administration of AMR extract. The results provided a meaningful basis for evaluating the clinical applications of traditional Chinese medicine. Copyright © 2012 John Wiley & Sons, Ltd.     

Preparative isolation and purification of atractylon and atractylenolide III from the Chinese medicinal plant atractylodes macrocephala by high-speed counter-current chromatography

The medicinal plant Atractylodes macrocephala (Baizhu in Chinese) has been widely used in traditional Chinese medicine for energy and stomach complaints, treatment of dyspepsia and anorexia, anti-inflammation, anticancer and for increasing assimilation. A high-speed counter-current chromatography (HSCCC) method was developed for the preparative separation and purification of two main bioactive components, namely, atractylon and atractylenolide III from A. macrocephala by using light petroleum (60-90 degrees C)-ethyl acetate-ethanol-water (4:1:4:1 v/v) as the two-phase solvent system in dual-mode elution. Compared with the separation using the normal-mode elution, the dual-mode HSCCC can be achieved with shorter elution time. Atractylenolide III (32.1 mg) at 99.0% purity and 319.6 mg atractylon at 97.8% purity could be obtained from 1000 mg crude sample in a single run. The recoveries of atractylenolide III and atractylon were 95.4 and 92.6%, respectively.     

白术挥发油中苍术酮氧化反应的动力学

白术挥发油;苍术酮;动力学方程;活化能;反应机理     

北苍术超临界CO2萃取产物的成分

用气相色谱－质谱－计算机联用法分析了北苍术的超临界CO2萃取产物的成分,鉴定出苍术酮、β－桉叶油醇、苍术醇31化合物,占出峰总面积的88．1％,并与传统水蒸汽蒸馏法（SD）提取的挥发油成分进行了对比。结果证明,两种方法抽提的苍术主成分基本相同,但超临界CO2萃取产物比SD产物多出现了一些重质组分,萃取更完全而有效。