羌活属一新种

本文记载了中国伞形科特有属羌活属１新种细叶羌活     

重订羌活属的分类

在野外实地观察和标本整理的基础上,检查了羌活属各个分类群的分类问题,考证了它们的名称。通过性状分析,主要以小总苞片的形状及其相关特征作为属下分类的依据,将卵叶羌活N．oviforme 改隶于宽叶羌活之下作为亚种处理,支持张盍曾在1975年将 N.franchetii 与N.forbesii合并为1种。如此,该属共5种,其中1种含1亚种。描述了两个新组,对属的特征和地理分布作了补充。近年该属新增加2种,因此分种检索表也作了相应的修改。     

A reclassification of Notopterygium Boissieu (Umbelliferae)

The genus Notopterygium is endemic to China. The plants of this genus are important traditional Chinese medicine. When established by H. de Boissieu in 1903, Notopterygium included only two species, i.e.N. franchetii and N. forbesii. Within nearly a century, five more specific names had been added to this genus, i.e.N. forrestii H. Wolff, N. oviforme Shan, N. incisum Ting ex H. T. Chang, N. pinnatiinvolucellatum Pu et Y. P. Wang, N. tenuifolium Sheh et Pu. Based on field ervation and examination of herbarium specimens, all the taxa in this genus were taxonomically reviewed and their nomenclature was carefully checked. N. oviforme was treated as a subspecies under N. forbesii. We agree with Chang He-Tseng in reducing N. franchetii to N.forbesii as a synonym. As a result, five species, one of which contains a subspecies, are recognized. Based on the morphology of involucel bractlets as well as their relevant characters, Notopterygium is divided into two sections.Sect. 1. Notopterygium Basal and proximal cauline leaves 2-ternate to 2 ～3-ternate-pinnate, ultimate divisions broadovate or ovate-lanceolate; involucel bractlets linear, entire, vascular bundles of petiole approximately equal in size. This section contains two species and one subspecies.Sect. 2. Tenuifolium Pu, sect. nov.Basal and proximal cauline leaves 2 ～3-ternate-pinnate, ultimate divisions ranceolate,ovate-lanceolate or linear; involucel bractlets linear, entire or oblanceolate, 2 ∽ 3-fid or pinnate at the apex; vascular bundles of petiole unequal in size. This section centains three species.     

羌活药材活性成分的提取工艺研究

选择提取次数、溶剂倍数、提取时间为考察因素,以提取浸膏量及三种主要活性物质(紫花前胡苷、异欧前胡素、羌活醇)含量为考察指标,探讨羌活药材活性成分的乙醇加热回流提取工艺。在首先考察单因素对提取效果影响的基础上采用正交试验法,优选出羌活药材活性成分的最佳提取工艺参数:加6倍量95%乙醇,加热回流提取3次,每次提取时间为1 h。     

羌活光合作用日变化及其与生理生态因子的关系

以大田引种栽培的羌活植株为材料,用Li-6400便携式光合作用测定系统原位测定自然条件下生长的羌活孕蕾期叶片的光合速率光响应、净光合速率及生理生态因子日变化,并通过相关分析、通径分析和逐步回归分析探讨净光合速率与生理生态因子的关系.结果表明:(1)羌活净光合速率(P_n)、 蒸腾速率(T_r)、气孔导度(G_s)日变化均呈双峰曲线,其净光合速率具有典型的"午休"现象,并主要由非气孔限制因素造成;(2)影响羌活叶片Pn日变化的主要决定生理因子是G_s,主要限制因子是胞间CO_2浓度(C_i);主要决定生态因子是空气相对湿度(RH),限制因子是气温(T_a); G_s是影响净光合速率最重要的生理生态因子.研究发现,羌活有较强的适应弱光环境的能力,属于阳性耐荫植物,宜选择海拔和荫蔽度均较高的环境进行引种驯化栽培,以利于其生长和存活.     

不同采收期栽培宽叶羌活挥发性成分的研究

采用水蒸气蒸馏法提取不同采收时间(5、6、7、8和9月)栽培宽叶羌活药材中的挥发油,测定其含量;通过GC-MS对挥发油成分进行了分析鉴定,并采用面积归一化法计算各组分的相对含量。实验结果表明,不同采收时间,栽培宽叶羌活挥发油含量存在差异,以8月份采收的药材挥发油含量最高;挥发油经GC-MS分析,共鉴定出39个化合物,有31种共有成分;对共有组分进行主成分分析显示,香桧烯、α-蒎烯、莰烯、β-蒎烯、γ-萜品烯、乙酸龙脑酯、α-红没药醇等15种成分可作为挥发油季节变化的特征组分。不同季节采集的羌活生药材,其挥发油含量和成分具有一定的差别,在一定程度上反映了其药用价值的微妙差异,可为羌活药材药理药用价值的进一步开发利用提供一定的参考。     

青海省不同地区羌活脂溶性化学成分的研究

采用毛细管气相-质谱法对青海省不同地区羌活脂溶性化学成分进行分析和鉴定,共分离出80余个成分,鉴定了60个,并用面积归一法确定其相对含量。结果表明,其化学成分以亚油酸、水菖蒲酮、十六烷酸及3,5-豆甾二烯为主,不同地区间羌活脂溶性化学成分种类和含量均有明显的差异。     

Analysis of sesquiterpene lactones in Eupatorium lindleyanum by HPLC‐PDA‐ESI‐MS/MS

Introduction – The aerial part Eupatorium lindleyanum is commonly used as an antipyretic and detoxicant clinically in traditional Chinese medicine. Our previous research showed that germacrane sesquiterpene lactones were its main active constituents, so the development of rapid and accurate methods for the identification of the sesquiterpene lactones is of great significance. Objective – To develop an HPLC‐PDA‐ESI‐MS/MS method capable for simple and rapid analysis of germacrane sesquiterpene lactones in the aerial part E. lindleyanum. Methodology – High‐performance liquid chromatography‐photodiode array detection‐electrospray ionization‐tandem mass spectrometry was used to analyze germacrane sesquiterpene lactones of Eupatorium lindleyanum. The fragmentation behavior of germacrane sesquiterpene lactones in a Micromass Q/TOF Mass Spectrometer was discussed, and 9 germacrane sesquiterpene lactones were identified by comparison of their characteristic data of HPLC and MS analyses with those obtained from reference compounds. Results – The investigated germacrane sesquiterpene lactones were identified as eupalinolides C (1), 3β‐acetoxy‐8β‐(4′‐hydroxy‐tigloyloxy)‐14‐hydroxy‐costunolide (2), eupalinolides A (3), eupalinolides B (4), eupalinolides E (5), 3β‐acetoxy‐8β‐(4′‐oxo‐tigloyloxy)‐14‐hydroxy‐heliangolide (6), 3β‐acetoxy‐8β‐(4′‐oxo‐ tigloyloxy)‐14‐hydroxy‐costunolide (7), hiyodorilactone B (8), and 3β‐acetoxy‐8β‐(4′‐hydroxy‐tigloyloxy)‐ costunolide (9). Compounds 6, 7 and 9 were reported for the first time. Conclusion – HPLC‐PDA‐ESI‐MS/MS provides a new powerful approach to identify germacrane sesquiterpene lactones in E. lindleyanum rapidly and accurately. Copyright © 2009 John Wiley & Sons, Ltd.     

Analysis of the rat hypothalamus proteome by data‐independent label‐free LC‐MS/MS

Studies of neuronal, endocrine, and metabolic disorders would be facilitated by characterization of the hypothalamus proteome. Protein extracts prepared from 16 whole rat hypothalami were measured by data‐independent label‐free nano LC‐MS/MS. Peptide features were detected, aligned, and searched against a rat Swiss‐Prot database using ProteinLynx Global Server v.2.5. The final combined dataset comprised 21 455 peptides, corresponding to 622 unique proteins, each identified by a minimum of two distinct peptides. The majority of the proteins (69%) were cytosolic, and 16% were membrane proteins. Important proteins involved in neurological and synaptic function were identified including several members of the Ras‐related protein family and proteins involved in glutamate biosynthesis.     

Analysis of flumequine enantiomers in rat plasma by UFLC‐ESI‐MS/MS

In this study the analysis and confirmation of flumequine enantiomers in rat plasma by ultra‐fast liquid chromatography coupled with electron spray ionization mass spectrometry (using propranolol as an internal standard [IS]) was developed and validated. Plasma samples were prepared by liquid–liquid extraction using methyl tert‐butyl ether as the extraction solvent. Direct resolution of the R‐ and S‐isomers was performed on a CHIRALCEL OJ‐RH column (4.6 × 150 mm, 5 μm) using acetonitrile / 0.1% formic acid / 1 mM ammonium acetate as the mobile phase. Detection was operated by electron spray ionization in the selected ion monitoring and positive ion mode. The target ions at m/z 262.1 and m/z 260.1 were selected for the quantification of the enantiomers and IS, respectively. The linear range was 0.5–500 ng/mL. The precisions (coefficient of variation, CV%) and recoveries were 1.43–8.68 and 94.24–106.76%, respectively. The lowest quantitation limit for both enantiomers is 0.5 ng/mL, which is sensitive enough to be applied to sample analysis in other related studies.     

MS2‐Deisotoper: A Tool for Deisotoping High‐Resolution MS/MS Spectra in Normal and Heavy Isotope‐Labelled Samples

High‐resolution MS/MS spectra of peptides can be deisotoped to identify monoisotopic masses of peptide fragments. The use of such masses should improve protein identification rates. However, deisotoping is not universally used and its benefits have not been fully explored. Here, MS2‐Deisotoper, a tool for use prior to database search, is used to identify monoisotopic peaks in centroided MS/MS spectra. MS2‐Deisotoper works by comparing the mass and relative intensity of each peptide fragment peak to every other peak of greater mass, and by applying a set of rules concerning mass and intensity differences. After comprehensive parameter optimization, it is shown that MS2‐Deisotoper can improve the number of peptide spectrum matches (PSMs) identified by up to 8.2% and proteins by up to 2.8%. It is effective with SILAC and non‐SILAC MS/MS data. The identification of unique peptide sequences is also improved, increasing the number of human proteoforms by 3.7%. Detailed investigation of results shows that deisotoping increases Mascot ion scores, improves FDR estimation for PSMs, and leads to greater protein sequence coverage. At a peptide level, it is found that the efficacy of deisotoping is affected by peptide mass and charge. MS2‐Deisotoper can be used via a user interface or as a command‐line tool.     

iTRAQ‐coupled 2‐D LC‐MS/MS analysis of protein profile associated with HBV‐modulated DNA methylation

The development of hepatocellular carcinoma (HCC) is believed to be associated with multiple risk factors, including the infection of hepatitis B virus (HBV). Based on the analysis of individual genes, evidence has indicated the association between HCC and HBV and has also been expanded to epigenetic regulation, with an involvement of HBV in the DNA methylation of the promoter of cellular target genes leading to changes in their expression. Proteomic study has been widely used to map a comprehensive protein profile, which in turn could provide a better understanding of underlying mechanisms of disease onset. In the present study, we performed a proteomic profiling by using iTRAQ‐coupled 2‐D LC/MS‐MS analysis to identify cellular genes down‐regulated in HBV‐producing HepG2.2.15 cells compared with HepG2 cells. A total of 15 proteins including S100A6 and Annexin A2 were identified by our approach. The significance of these cellular proteins as target of HBV‐mediated epigenetic regulation was supported by our validation assays, including their reactivation in cells treated with 5‐aza‐2′‐deoxycytidine (a DNA methyltransferase inhibitor) by real‐time RT‐PCR and Western blot analysis, as well as the DNA methylation status analysis by bisulfite genome sequencing. Our approach provides a comprehensive analysis of cellular target proteins to HBV‐mediated epigenetic regulation and further analysis should facilitate a better understanding of its involvement in HCC development.     

Targeted On‐line SPE‐LC‐MS/MS Assay for the Quantitation of 12 Apolipoproteins from Human Blood

Laborious sample pretreatment of biological samples represents the most limiting factor for the translation of targeted proteomics assays from research to clinical routine. An optimized method for the simultaneous quantitation of 12 major apolipoproteins (apos) combining on‐line SPE and fast LC‐MS/MS analysis in 6.5 min total run time was developed, reducing the manual sample pretreatment time of 3 μL serum or plasma by 60%. Within‐run and between‐day imprecisions below 10 and 15% (n = 10) and high recovery rates (94–131%) were obtained applying the high‐throughput setup. High‐quality porcine trypsin was used, which outperformed cost‐effective bovine trypsin regarding digestion efficiency. Comparisons with immunoassays and another LC‐MS/MS assay demonstrated good correlation (Pearson's R: 0.81–0.98). Further, requirements on sample quality concerning sampling, processing, and long‐term storage up to 1 year were investigated revealing significant influences of the applied sampling material and coagulant on quantitation results. Apo profiles of 1339 subjects of the LIFE‐Adult‐Study were associated with lifestyle and physiological parameters as well as establish parameters of lipid metabolism (e.g., triglycerides, cholesterol). Besides gender effects, most significant impact was seen regarding lipid‐lowering medication. In conclusion, this novel highly standardized, high‐throughput targeted proteomics assay utilizes a fast, simultaneous analysis of 12 apos from least sample amounts.     

LFQuant: A label‐free fast quantitative analysis tool for high‐resolution LC‐MS/MS proteomics data

Database searching based methods for label‐free quantification aim to reconstruct the peptide extracted ion chromatogram based on the identification information, which can limit the search space and thus make the data processing much faster. The random effect of the MS/MS sampling can be remedied by cross‐assignment among different runs. Here, we present a new label‐free fast quantitative analysis tool, LFQuant, for high‐resolution LC‐MS/MS proteomics data based on database searching. It is designed to accept raw data in two common formats (mzXML and Thermo RAW), and database search results from mainstream tools (MASCOT, SEQUEST, and X!Tandem), as input data. LFQuant can handle large‐scale label‐free data with fractionation such as SDS‐PAGE and 2D LC. It is easy to use and provides handy user interfaces for data loading, parameter setting, quantitative analysis, and quantitative data visualization. LFQuant was compared with two common quantification software packages, MaxQuant and IDEAL‐Q, on the replication data set and the UPS1 standard data set. The results show that LFQuant performs better than them in terms of both precision and accuracy, and consumes significantly less processing time. LFQuant is freely available under the GNU General Public License v3.0 at http://sourceforge.net/projects/lfquant/ .     

Potential distribution of Notopterygium incisum Ting ex H. T. Chang and its predicted responses to climate change based on a comprehensive habitat suitability model

Notopterygium incisum Ting ex H. T. Chang is a rare and endangered traditional Chinese medicinal plant. In this research, we built a comprehensive habitat suitability (CHS) model to analyze the potential suitable habitat distribution of this species in the present and future in China. First, using nine different algorithms, we built an ensemble model to explore the possible impacts of climate change on the habitat distribution of this species. Then, based on this model, we built a CHS model to further identify the distribution characteristics of N. incisum‐suitable habitats in three time periods (current, 2050s, and 2070s) while considering the effects of soil and vegetation conditions. The results indicated that the current suitable habitat for N. incisum covers approximately 83.76 × 10³ km², and these locations were concentrated in the Tibet Autonomous Region, Gansu Province, Qinghai Province, and Sichuan Province. In the future, the areas of suitable habitat for N. incisum would significantly decrease and would be 69.53 × 10³ km² and 60.21 × 10³ km² in the 2050s and 2070s, respectively. However, the area of marginally suitable habitat would remain relatively stable. This study provides a more reliable and comprehensive method for modelling the current and future distributions of N. incisum, and it provides valuable insights for highlighting priority areas for medicinal plant conservation and resource utilization.