Phytochemical Analysis

Cover image for Vol. 23 Issue 4

July/August 2012

Volume 23, Issue 4

Pages 285–420

  1. Research Articles

    1. Top of page
    2. Research Articles
    1. Enhanced Sample Preparation for Quantitation of Microcystins by Matrix-Assisted Laser Desorption/Ionisation–Time of Flight Mass Spectrometry (pages 285–291)

      Jonathan Puddick, Michèle R. Prinsep, Susanna A. Wood, S. Craig Cary and David P. Hamilton

      Article first published online: 29 SEP 2011 | DOI: 10.1002/pca.1356

      Quantitative analysis of microcystins by matrix-assisted laser desorption/ionisation-time of flight (MALDI-TOF) mass spectrometryhas the potential to process large numbers of samples quickly and economically. The existing method uses an expensive internal standard and protocols that are incompatible with automated sample preparation and data acquisition. Seven different MALDI-TOF sample preparations were assessed for signal reproducibility and sensitivity using a cost-effective internal standard. One of these preparations was suitable for use in an automated work-flow, while maintaining reproducibility and sensitivity.

    2. Optimisation of Infrared-assisted Extraction of Rutin from Crude Flos Sophorae Immaturus Using Response Surface Methodology and HPLC Analysis (pages 292–298)

      Fa-jie Li, Su-li Ning, Yan Li, Ying-jia Yu, Ci-dan Shen and Geng-li Duan

      Article first published online: 18 OCT 2011 | DOI: 10.1002/pca.1357

      Rutin, one of main constitutes in Flos Sophorae Immaturus, has been proved to possess several pharmacological properties. However, optimization of the extraction of rutin from Flos Sophorae Immaturus is scarcely reported. Thus, it is significant to develop an effective method to obtain maximum yield of rutin from Flos Sophorae Immaturus. The present study demonstrated the feasibility of response surface methodology in the optimization of infrared-assisted extraction of rutin from crude Flos Sophorae Immaturus.

    3. Development of a Rapid Resolution Liquid Chromatographic Method Combined with Chemometrics for Quality Control of Angelicae dahuricae radix (pages 299–307)

      Gang Fan, Rui Deng, Lin Zhou, Xianli Meng, Tingting Kuang, Xianrong Lai, Jing Zhang and Yi Zhang

      Article first published online: 29 SEP 2011 | DOI: 10.1002/pca.1358

      A rapid and sensitive method using rapid resolution liquid chromatography combined with chemometrics was developed for differentiating the sulfur-fumigated and sun-dried Angelicae dahuricae radix samples with the aim of quality control. The results showed that sulphur-fumigation obviously destroyed the phytochemical profiles of Angelicae dahuricae radix. In addition, the critical markers including oxypeucedanin and imperatorin were ultimately screened out based on principal component analysis, which could be used for quality control of Angelicae dahuricae radix in the future.

    4. Eco-friendly separation of catechins using cyclodextrins as mobile phase additives in RP-HPLC (pages 308–314)

      Wentao Bi, Shengnan Li and Kyung Ho Row

      Article first published online: 13 OCT 2011 | DOI: 10.1002/pca.1359

      1. New mobile phases for RP-HPLC were developed for the separation of catechin compounds in tea.
      2. Eco-friendly cyclodextrins were used as mobile phase additives to decrease the proportion of organic modifier and improve resolutions and retention factors.
      3. Cyclodextrins can separate analytes through host–guest complexation, where a transient diastereomeric complex is formed between the cyclodextrin and the analyte.
    5. Screening and Determination for Potential α-Glucosidase Inhibitors from Leaves of Acanthopanax senticosus Harms by Using UF-LC/MS and ESI-MSn (pages 315–323)

      Hui Zhou, Junpeng Xing, Shu Liu, Fengrui Song, Zongwei Cai, Zifeng Pi, Zhiqiang Liu and Shuying Liu

      Article first published online: 26 SEP 2011 | DOI: 10.1002/pca.1360

      UF-LC/MS screening assay was developed for screening candidates of α-glucosidase inhibitors from leaves of Acanthopanax Senticosus Harms. Eight compounds that might bind to α-glucosidase were separated and detected. Subsequently, seven ligands were successfully identified. The α-glucosidase inhibition activity of the related compounds in leaves of Acanthopanax Senticosus Harms was determined by using in vitro assays. The comparison of the α-glucosidase inhibitory activity indicated that glycosylation of the flavonol quercetin on ring C exhibited a decreased effect on the α-glucosidase inhibition activity, while the caffeoyl group attached to a quinic acid core enhanced the α-glucosidase inhibition activity.

    6. Simultaneous Determination of Sulphur metabolites in Arabidopsis thaliana via LC-ESI-MS/MS and 34S-metabolic Labelling (pages 324–331)

      Chin-Lin Hsieh, Kai-Wun Yeh, Luit J. De Kok, Ryh-Nan Pan, Yueh-Hsiung Kuo and Mei-Hwei Tseng

      Article first published online: 10 OCT 2011 | DOI: 10.1002/pca.1361

      An analytical method based on liquid chromatography, electrospray ionisation, tandem mass spectrometry (LC-ESI-MS/MS) and 34S-metabolic ([34S]Met) labelling was developed for the accurate quantification of Met, glutathione disulphide (GSSG), and glutathione (GSH) contents in Arabidopsis thaliana tissues. [34S]GSSG, [34S]Met and [34S]GSH were biosynthesised in the A. thaliana plant, which was grown using Na234SO4-containing media. These 34S-labelled metabolites were applied as internal standards (ISs) for the analysis and quantification of endogenous sulphur metabolites in root and shoot extracts of A. thaliana.

    7. Preparative Separation of Methylswertianin, Swerchirin and Decussatin from the Tibetan Medicinal Plant Swertia Mussotii Using High-speed Counter-current Chromatography (pages 332–336)

      Jing Jia, Tao Chen, Ping Wang, Guichen Chen, Jinmao You, Yongjun Liu and Yulin Li

      Article first published online: 3 NOV 2011 | DOI: 10.1002/pca.1362

      A high performance and environmentally, friendly method for the preparative separation of three xanthones including methylswertianin, swerchirin and decussatin from Swertia mussotii by using HSCCC was established. The solvent system of HSCCC was composed of n-hexane–ethyl acetate–methanol –water (5:5:10:4, v/v/v/v). Using the separated method of HSCCC, 8 mg of methylswertianin, 21 mg of swerchirin and 11 mg of decussatin with purities of over 98% could be isolated from a 150 mg crude sample.

    8. Quantitative and Transformation Product Analysis of Major Active Physalins from Physalis Alkekengi Var. Franchetii (Chinese Lantern) Using Ultraperformance Liquid Chromatography with Electrospray Ionisation Tandem Mass Spectrometry and Time-of-flight Mass Spectrometry (pages 337–344)

      Yunliang Zheng, Yong Chen, Yiping Ren, Lianjun Luan and Yongjiang Wu

      Article first published online: 13 OCT 2011 | DOI: 10.1002/pca.1363

      A separation and quantification method based on ultra-performance liquid chromatography with electrospray ionization tandem mass spectrometry detection (UPLC-ESI-MS/MS) was developed for the quantitative analysis of five active physalins (physalin B, physalin D, physalin G, physalin H and 4,7-didehydroneophysalin B) in Chinese lantern. The separation was carried out on an Acquity UPLC BEH Shield RP C18 column with water and acetonitrile as the mobile phase under gradient condition. All the compounds showed good linearity (R2>0.998). The recoveries, measured at three concentration levels, varied from 98.8 to 101.4% with RSDs <4.5%. The simple, rapid and reliable method was successfully applied to quantify the five components in 14 samples of Chinese lantern from different areas. The transformation products of 4,7-didehydroneophysalin B was studied for the first time and three transformation products were detected by UPLC-PDA-ESI-MS/MS and identified through comparing their HRMS and MS2 ion fragmentations with corresponding references.

    9. Establishment and Application of a Metabolomics Workflow for Identification and Profiling of Volatiles from Leaves of Vitis vinifera by HS-SPME-GC-MS (pages 345–358)

      Georg Weingart, Bernhard Kluger, Astrid Forneck, Rudolf Krska and Rainer Schuhmacher

      Article first published online: 18 OCT 2011 | DOI: 10.1002/pca.1364

      We developed a metabolomics workflow for the automated extraction, measurement and identification of volatiles in plant leaves. Harvested samples were quenched (liquid nitrogen), homogenised and analysed by HS-SPME-GC-MS. Data were processed with AMDIS. For positive annotation of metabolites, MS spectral match factor had to be ≥ = 90 and relative RI deviation ≤ 2 % on two stationary GC phases. Application to field samples of grapevine leaves resulting in the annotation of 63 metabolites, of which 47 were confirmed with authentic standards.

    10. Chemical-based Species Classification of Rhubarb Using Simultaneous Determination of Five Bioactive Substances by HPLC and LDA Analysis (pages 359–364)

      Chu VanMen, Yu Seon Jang, Hong Mei Zhu, Jae Hyun Lee, Trinh Nam Trung, Tran Minh Ngoc, Young Ho Kim and Jong Seong Kang

      Article first published online: 18 OCT 2011 | DOI: 10.1002/pca.1365

      Rhubarb is a traditional Chinese medicine derived from the rhizome of three species: Rheum tanguticum, Rheum palmatum and Rheum officinale. There are several species that are often misidentified as rhubarb. Taxonomical identification of these various species can be challenging. A chemical-based species classification method of rhubarb using simultaneous determination of bioactive compounds by HPLC has been developed with 39 known samples of five different species and successfully applied to identify 57 unknown samples collected from Korea and China.

    11. Quality Evaluation of a Herbal Prescription Through Quantification of 40 Components by HPLC–ESI–MS/MS (pages 365–372)

      Man Liu, Songchen Liu, Xiaowei Shi, Minyan Liu, Yingfeng Du’ Lantong Zhang and Qiao Wang

      Article first published online: 21 OCT 2011 | DOI: 10.1002/pca.1366

      The Kang-nao-shuai (KNS) capsule is a combined herbal prescription. A HPLC-ESI-MS method for the quality evaluation of KNS capsules through quantification of 40 components was established. Most of the 40 components were determined for the first time in the KNS capsule. And the method was successfully applied to analyse 10 batches of KNS capsule.

    12. Determination of Nucleosides and Nucleobases in the Pollen of Typha Angustifolia by UPLC-PDA-MS (pages 373–378)

      Wei-Wei Tao, Jin-Ao Duan, Nian-Yun Yang, Sheng Guo, Zhen-Hua Zhu, Yu-Ping Tang and Da-Wei Qian

      Article first published online: 24 OCT 2011 | DOI: 10.1002/pca.1367

      In this study, ten nucleosides and nucleobases in the pollen of Typha angustifolia were simultaneously determined by UPLC-PDA. Furthermore, UPLC coupled with an electrospray ionization tandem mass spectrometer (ESI-MS/MS) method was used to further confirm the identity of the target compounds by comparing their MS data with those of reference standards. The proposed method was successfully applied to the analysis of 15 samples collected from different places in China.

    13. Comparison of Headspace Solid-phase Microextraction, Headspace Single-drop Microextraction and Hydrodistillation for Chemical Screening of Volatiles in Myrtus Communis L (pages 379–386)

      Mohammad Moradi, Massoud Kaykhaii, Ali Reza Ghiasvand, Shahriar Shadabi and Alinazar Salehinia

      Article first published online: 8 NOV 2011 | DOI: 10.1002/pca.1368

      Three methods of headspace solid phase microextraction (HS-SPME), headspace single-drop microextraction (HS-SDME) and hydrodistillation were used for the extraction of volatile compounds of Myrtus communis followed by GC-MS analysis. SPME and SDME parameters including extraction temperature, amount and size of ground sample, extraction time and sample vial volume were optimized. The number of identified compounds was 22 (96.9%) for HD, 20 (98.1%) for HS-SPME and 18 (98.9%) for HS-SDME.

    14. Quality Assessment of Commercial Magnoliae Officinalis Cortex by 1H-NMR-based Metabolomics and HPLC Methods (pages 387–395)

      Yong Jiang, Julie Vaysse, Véronique Gilard, Stéphane Balayssac, Sébastien Déjean, Myriam Malet-Martino, Bruno David, Christel Fiorini and Yves Barbin

      Article first published online: 24 OCT 2011 | DOI: 10.1002/pca.1369

      A 1H-NMR-based metabolomics was developed along with the traditional HPLC method for the quality control of commercial Magnoliae officinalis Cortex. The different varieties, different medicinal parts and different qualities of Magnoliae officinalis Cortex could be differentiated from 1H-NMR data, and honokiol and magnolol were found to be main compounds responsible for the discrimination, which proves that the choice of these two compounds as markers for HPLC to assess the quality of Magnoliae officinalis Cortex is relevant.

    15. Analysis of Cardamonin by Square Wave Voltammetry (pages 396–399)

      Ana M. Carvalho, Luís M. Gonçalves, Inês M. Valente, José A. Rodrigues and Aquiles A. Barros

      Article first published online: 24 OCT 2011 | DOI: 10.1002/pca.1370

      Cardamonin was analysed electrochemically by means of a hanging mercury drop electrode (HMDE) using square wave voltammetry (SWV). It was extracted from cardamom spice and quantified thereafter using the standard additions method to overcome matrix effects. Decoction using ethanol as the extraction solvent appears to be the simplest extraction technique. The LOD and LOQ of the proposed voltammetric methodology are adequate for trace analysis of this compound in several phytochemical matrices.

    16. Effects of Auxins on the Production of Steroidal Alkaloids in Rapidly Proliferating Tissue and Cell Cultures of Solanum lyratum (pages 400–404)

      Chung-Io Kuo, Chi-Hsein Chao and Mei-Kuang Lu

      Article first published online: 19 OCT 2011 | DOI: 10.1002/pca.1371

      Solanum lyratum, a rare species, is used to treat cancer, tumors and warts. Plant cell- and tissue-culture of S. lyratum, producing steroidal alkaloids, could be useful supplements to natural sources. We previously showed that a culture system of its cells could produce steroidal alkaloids. The purpose of this study was to study the production of solanine, solanidine, and solasodine by adding auxin-type phytohormones including indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), naphthaleneacetic acid (NAA), and 2,4-dichlorophenoxyacetic acid (2,4-D) to cell and callus cultures of this species. 2,4-D-induced calli from roots led to a significant enhancement in solanine production with a value of 4.13 mg/g dry weight (DW). The maximal solanidine and solasodine levels of 6.26 and 7.69 mg/g DW were respectively obtained with IBA- and IAA-treated S. lyratum cells at concentrations of 1 and 5 mg/l.

    17. TLC Bioautographic Method for Detecting Lipase Inhibitors (pages 405–407)

      Abdel Moniem Sadek Hassan

      Article first published online: 17 NOV 2011 | DOI: 10.1002/pca.1372

      A rapid bioautographic enzyme assay has been developed for the screening of lipase inhibition by plant extracts. Enzyme activity was detected by conversion of naphthyl acetate into naphthol, which reacts with fast blue b salt to form a purple -colored diazonium dye. A positive result could be visualized directly as white spot(s) against a dye-colored background. The technique is sensitive enough to give a clear result in the the presence of only 0.01 µg of orlistat.

    18. Simultaneous Determination of Four Sesquiterpenoids in Atractylodes Macrocephala Rhizoma by GC-FID: Optimisation of an Ultrasound-assisted Extraction by Central Composite Design (pages 408–414)

      Yang-Yang Shi, Shu-Hong Guan, Ren-Neng Tang, Si-Jia Tao and De-An Guo

      Article first published online: 17 NOV 2011 | DOI: 10.1002/pca.1373

      A validated gas chromatography coupled with flame ionisation detector method was developed to simultaneous determine four sesquiterpenoids in Atractylodes Macrocephala Rhizoma. In order to increase extraction efficiency, central composite design was conducted to optimize ultrasound-assisted extraction parameters. The result showed that the developed GC-FID method was feasible for quality control of AMR and UAE under optimum conditions could be considered as a powerful tool for the extraction of phytochemicals from plants.

    19. LC-UV Assay Method and UPLC/Q-TOF-MS Characterisation of Saponins from Ilex Paraguariensis A. St. Hil. (Mate) Unripe Fruits (pages 415–420)

      Maria Paula Garofo Peixoto, Samuel Kaiser, Simone Gasparin Verza, Pedro Ernesto de Resende, Janine Treter, Cabral Pavei, Gustavo Luís Borré and George González Ortega

      Article first published online: 21 NOV 2011 | DOI: 10.1002/pca.1374

      In this work a mate (Ilex paraguariensis) saponin fraction was obtained. A LC-UV method to assay the total content of saponins in unripe mate fruits was validated and ilexoside II was used as external standard. The chemical structures of 11 triterpenic saponins were characterized by UPLC/Q-TOF-MS. The total saponin content determined in unripe fruits was 7.28 % (w/w) and the saponins present in mate saponin fraction are derived from ursolic/oleanolic, acetyl ursolic or pomolic acid, mainly.