Phytochemical Analysis

Cover image for Vol. 24 Issue 6

November/December 2013

Volume 24, Issue 6

Pages 513–688

  1. Research Articles

    1. Top of page
    2. Research Articles
    1. A Rapid Method for Total Β-Escin Analysis in Dry, Hydroalcoholic and Hydroglycolic Extracts of Aesculus hippocastanum L. by Capillary Zone Electrophoresis (pages 513–519)

      Lidiane S. Dutra, Magda N. Leite, Marcos A. F. Brandão, Priscila Aparecida de Almeida, Fernando A. S. Vaz and Marcone A. L. de Oliveira

      Version of Record online: 18 MAR 2013 | DOI: 10.1002/pca.2425

      A capillary zone electrophoresis method capable to determine, in 6 minutes, total β-escin in different extracts of horse chestnut (Aesculus hippocastanum L.) was developed, optimized and validated through chemometric approaches. An electrolyte, composed of bicarbonate-carbonate buffer (pH 10.3), was used with the conditions: +20 kV of voltage; 20 °C of temperature; direct ultraviolet detection at 226 nm; 13 mbar injection for 5 s. This method was applied on these samples demonstrating its usefulness to analysis of this plant species.

    2. A Modern Approach to the Authentication and Quality Assessment of Thyme Using UV Spectroscopy and Chemometric Analysis (pages 520–526)

      Haidy A. Gad, Sherweit H. El-Ahmady, Mohamed I. Abou-Shoer and Mohamed M. Al-Azizi

      Version of Record online: 10 MAR 2013 | DOI: 10.1002/pca.2426

      Thyme represents a medicinal herb that is constantly adulterated due to its resemblance to many plants with similar organoleptic properties. This study presents a simple model for the quality assessment of Thymus species using UV spectroscopy together with chemometric techniques: PCA, SIMCA and HCA. The model was able to discriminate T. vulgaris from other species, all traded in the Egyptian market as different types of thyme as well as discriminate closely related species and finally classify commercial varieties into thyme or non-thyme.

    3. Simultaneous Determination of Five Bioactive Components in Radix Glycyrrhizae by Pressurised Liquid Extraction Combined with UPLC–PDA and UPLC/ESI–QTOF–MS Confirmation (pages 527–533)

      Shujun Zhou, Jiliang Cao, Feng Qiu, Weijun Kong, Shihai Yang and Meihua Yang

      Version of Record online: 21 FEB 2013 | DOI: 10.1002/pca.2427

      A rapid and accurate UPLC–PDA method, based on pressurized liquid extraction (PLE), is proposed to simultaneously quantify five bioactive compounds, namely liquiritin apioside, liquiritin, liquiritigenin, glycyrrhizic acid and glycyrrhetinic acid in 12 batches radix Glycyrrhizae samples from different origins in China. Furthermore, the samples were analyzed on an UPLC/ESI–Q–TOF–MS system to confirm the results. The established PLE and UPLC–PDA method could serve as a rapid and effective method for quality evaluation of Glycyrrhizae speices.

    4. Ellagic Acid and Derivatives from Cochlospermum angolensis Welw. Extracts: HPLC–DAD–ESI/MSn Profiling, Quantification and In Vitro Anti-depressant, Anti-cholinesterase and Anti-oxidant Activities (pages 534–540)

      Federico Ferreres, Clara Grosso, Angel Gil-Izquierdo, Patrícia Valentão and Paula B. Andrade

      Version of Record online: 1 APR 2013 | DOI: 10.1002/pca.2429

      This study reports for the first time the phenolic composition and neuroprotective effect of Cochlospermum angolensis Welw. bark. The HPLC-DAD-ESI/MSn analysis revealed the presence of eight compounds, ellagic acid, methyl ellagic acid and their derivatives. The testes extracts and ellagic acid exhibited strong radical scavenging activity and antidepressant potential, but a weak effect against acetylcholinesterase and butyrylcholinesterase. These results provide evidence of the value of C. angolensis as source of health-promoting antioxidants and antidepressant compounds.

    5. Characterisation of Homoflavonoids from Three Ophioglossum Species Using Liquid Chromatography with Diode Array Detection and Electrospray Ionisation Tandem Mass Spectrometry (pages 541–549)

      Chuan-Xing Wan, Jian-Guang Luo, Yu-Cheng Gu, De-Ran Xu and Ling-Yi Kong

      Version of Record online: 12 APR 2013 | DOI: 10.1002/pca.2430

      A rapid method for identifying homoflavonoids from Ophioglossum by using HPLC–DAD–MSn was developed according to the analysis of their typical MS fragmentation and UV characteristics. The analyses of the MS/MS spectra of the homoflavonoids allowed us to classify them into two types according to their fragmentation characteristics. The type I homoflavonoids, with an attached additional carbon atom to the C-3 position of the C-ring, presented the initial competing loss of H2O and CH2O from their aglycone ions, compared to the initial removal of H2O or CO in the case of the type II homoflavonoids, that bear the additional carbon atom at the C-2' site of the B-ring and forming ring D. The above characteristic fragmentations of homoflavonoids were quite different from those of other flavonoids, and were successfully used to identify homoflavonoids from the crude extracts of three Ophioglossum species.

    6. A Novel Approach in Herbal Quality Control Using Hyperspectral Imaging: Discriminating Between Sceletium tortuosum and Sceletium crassicaule (pages 550–555)

      Emmanuel Amukohe Shikanga, Alvaro M. Viljoen, Ilze Vermaak and Sandra Combrinck

      Version of Record online: 17 APR 2013 | DOI: 10.1002/pca.2431

      Sceletium tortuosum is the most popular species of the genus Sceletium, but displays several phytochemical and morphological similarities to S. crassicaule. Although chromatography is widely used for quality monitoring of phytomedicines, a combination of hyperspectral imaging and chemometrics proved to be superior for differentiating S. tortuosum from S. crassicaule specimens.

    7. Identification and Characterisation of Phenolics in Polygonum capitatum by Ultrahigh-Performance Liquid Chromatography with Photodiode Array Detection and Tandem Mass Spectrometry (pages 556–568)

      Shang-Gao Liao, Li-Juan Zhang, Fa Sun, Zhen Wang, Xun He, Ai-Min Wang, Yong-Jun Li, Yong Huang, Yan-Yu Lan, Bo-Li Zhang and Yong-Lin Wang

      Version of Record online: 23 MAY 2013 | DOI: 10.1002/pca.2432

      Polygonum capitatum is a well-known Chinese medicinal plant widely used by the Miao people for the treatment of various urologic disorders. The use of UPLC–PDA–MS/MS has led to the identification or structural characterization of an amino acid and 40 phenolics including a number of known and unknown flavonoid glycosides and polyphenolic glycosides in a tannin-free phenolic fraction. Results showed that polyphenolic glycosides were perhaps the most typical chemical markers of P. capitatum.

    8. A Simple and Rapid Single Kernel Screening Method to Estimate Amylose Content in Rice Grains (pages 569–573)

      Somanath Agasimani, G. Selvakumar, A. John Joel and Sundaram Ganesh Ram

      Version of Record online: 1 APR 2013 | DOI: 10.1002/pca.2433

      The Presently used standard protocols for the estimation of Amylose content (AC) are laborious and costly. A rapid screening protocol based on the amylose-iodine complex which uses time duration for colouration of the cut grain portion has been developed and standardized. Perfect linear relationship between the time taken for colouration by the cut grain dip (CGD) method and the AC values estimated through routine spectrophotometric method validated the efficiency of this method.

    9. Analysis of Cleistopholis patens Leaf and Trunk Bark Oils Using Combined GC- Flame Ionisation Detection, GC-Retention Index, GC–MS and 13C-NMR (pages 574–580)

      Zana A. Ouattara, Jean Brice Boti, Antoine Coffy Ahibo, Joseph Casanova, Félix Tomi and Ange Bighelli

      Version of Record online: 17 APR 2013 | DOI: 10.1002/pca.2435

      A sample of leaf oil and two samples of trunk bark oil from Ivorian Cleistopholis patens were submitted to GC-FID, GC-RI, GC-MS and 13C-NMR analysis. The compositions were dominated (>10%) by (E)-β-caryophyllene/germacrene D, germacrene B/α-phellandrene, and germacrene D/β -elemol, respectively. The true content of thermolabile compounds, germacrene A, germacrene B and germacrene C and the rearranged products β-elemene, γ-elemene and δ-element was evaluated by combination of GC-FID and 13C-NMR data.

    10. Validation of a Generic Quantitative 1H NMR Method for Natural Products Analysis (pages 581–597)

      Tanja Gödecke, José G. Napolitano, María F. Rodríguez-Brasco, Shao-Nong Chen, Birgit U. Jaki, David C. Lankin and Guido F. Pauli

      Version of Record online: 5 JUN 2013 | DOI: 10.1002/pca.2436

      Quantitative 1H NMR (qHNMR) is a suitable technique for quality control (QC) of natural products (NP) and botanical dietary supplements (BDS). The presented generic qHNMR protocol provides guidance for both purity assessment of NP and quantitation of marker compounds in complex BDS. Several qHNMR calibration approaches and post-acquisition processing strategies were examined with respect to their impact on method validation. The study revealed that acquisition, processing parameters, and processing software are equally important for building validating qHNMR methods.

    11. A Proposed Biosynthetic Pathway of Picrosides Linked through the Detection of Biochemical Intermediates in the Endangered Medicinal Herb Picrorhiza kurroa (pages 598–602)

      Varun Kumar, Hemant Sood, Manu Sharma and Rajinder Singh Chauhan

      Version of Record online: 21 MAY 2013 | DOI: 10.1002/pca.2437

      LC-ESI-MS/MS, was used to assemble the complete picrosides biosynthetic pathway in P. kurroa through the detection of catalpol and aucubin, two major backbone structures of picrosides along with intermediate metabolites; boschnaloside, bartsioside and mussaenosidic acid, which were confirmed in ESI-negative mode with pseudo-molecular ion peaks i.e. m/z 361, m/z 343, m/z 345, m/z 329 and m/z 375 ions and their fragmentation patterns.

    12. One-step Separation and Purification of Three Lignans and One Flavonol from Sinopodophyllum emodi by Medium-pressure Liquid Chromatography and High-speed Counter-current Chromatography (pages 603–607)

      Ping Wang, Yongling Liu, Tao Chen, Wenhua Xu, Jinmao You, Yongjun Liu and Yulin Li

      Version of Record online: 30 MAY 2013 | DOI: 10.1002/pca.2438

      Lignans and flavonols are the primary constituents of Sinopodophyllum emodi and have been used as cathartic, anthelmintic, chemotherapeutic and anti-hypertensive compounds. In this paper, three lignans and one flavonol, including 4′-demethyl podophyllotoxin, podophyllotoxin, deoxypodophyllotoxin and kaempferol, were firstly enriched by medium-pressure liquid chromatography(MPLC) and then separated by high-speed counter-current chromatography(HSCCC) in one step using a solvent system composed of n-hexane: ethyl acetate: methanol: water (1.75:1.5:1:0.75, v/v/v/v).

    13. Chemical and Functional Characterisation of Propolis Collected from East Andalusia (Southern Spain) (pages 608–615)

      Shigenori Kumazawa, Josep Serra Bonvehí, Cristina Torres, Ahn Mok-Ryeon and Francisco José Orantes Bermejo

      Version of Record online: 14 MAY 2013 | DOI: 10.1002/pca.2439

      Propolis is a complex mixture of natural resinous components produced by honeybees from plant materials. The objectives of this study were to chemically characterize the compounds and evaluate the antioxidant activity found in twenty eight Andalusian propolis samples. All propolis samples had strong antioxidant activity, accompanied by high total phenolic content. The most abundant compounds were flavonoids. Concerning the phenolic compounds content, our results showed that the 75% of the samples analyzed contained at least 80 mg/g of flavonoids.

    14. Saponin Inventory from Argania spinosa Kernel Cakes by Liquid Chromatography and Mass Spectrometry (pages 616–622)

      Max Henry, Mariusz Kowalczyk, Mariateresa Maldini, Sonia Piacente, Anna Stochmal and Wiesław Oleszek

      Version of Record online: 18 JUN 2013 | DOI: 10.1002/pca.2440

      Argania spinosa kernel cakes, byproducts of argan oil extraction process, are known to contain large amounts of saponins. Only a few of them were characterized previously. This was due to the choice of pure ethanol as extracting solvent. The aqueous 50% ethanol improved drastically the extraction of more polar saponins. Four GOTCAB saponins (Glucuronide oleanane-type triterpene carboxylic acid 3,28-bidesmosides) were structurally identified by NMR analyses in this extract. LC-MS analyses showed the presence of at least 19 additional polar saponins.

    15. New and Highly Efficient Column Chromatographic Extraction and Simple Purification of Camptothecin from Camptotheca acuminata and Nothapodytes pittosporoides (pages 623–630)

      Xue-Hua Zeng, Yue-Hong Li, Shan-Shan Wu, Rui-Lin Hao, Hui Li, He Ni, Han-Bing Han and Hai-Hang Li

      Version of Record online: 30 MAY 2013 | DOI: 10.1002/pca.2441

      Camptothecin was extracted from leaves and fruits of Camptotheca acuminata and from roots of Nothapodytes pittosporoides at a 98% rate with 15- or 18-fold solvent for content analysis through a column chromatographic extraction precedure, or at a 97% rate with five- or sixfold solvent for production through a cyclic column chromatographic extraction. Extracts were purified to 98% through simple precipitation, fractionation and crystallization. The overall recovery rates of camptothecin from plant materials to purified products reached 92% or higher.

    16. 1H-NMR Quantification of Major Saccharides in Açaí Raw Materials: a Comparison of the Internal Standard Methodology with the Absolute Intensity qNMR Method (pages 631–637)

      Cole Sterling, Ronald Crouch, David J. Russell and Angela I. Calderón

      Version of Record online: 23 MAY 2013 | DOI: 10.1002/pca.2442

      In this study, the validation of Agilent's absolute intensity qNMR utility is evaluated with ibuprofen, and its application is compared against the traditional internal standard approach by assessing differences in the major saccharide components of three types of açaí raw materials: organic, freeze-dried non-organic and processed non-organic açaí powders. The comparable results generated from these two quantitative techniques indicated the accuracy, precision, and reliability of the absolute intensity qNMR utility as a tool for 1H-NMR quantification.

    17. Development and Validation Method for Simultaneous Quantification of Phenolic Compounds in Natural Extracts and Nanosystems (pages 638–644)

      Sara Baptista da Silva, Ana Oliveira, Domingos Ferreira, Bruno Sarmento and Manuela Pintado

      Version of Record online: 28 MAY 2013 | DOI: 10.1002/pca.2446

      This study shows a new HPLC method validated to simultaneous quantification of rosmarinic acid and quercetin in natural extracts and chitosan nanoparticles. This method is also able for detection and quantification of other phenolic and flavanoid compounds in natural matrix.

    18. Development of an Eastern Blotting Technique for the Visual Detection of Aristolochic Acids in Aristolochia and Asarum Species by Using a Monoclonal Antibody Against Aristolochic Acids I and II (pages 645–653)

      Xiao-Wei Li, Osamu Morinaga, Min Tian, Takuhiro Uto, Jie Yu, Ming-Ying Shang, Xuan Wang, Shao-Qing Cai and Yukihiro Shoyama

      Version of Record online: 12 JUN 2013 | DOI: 10.1002/pca.2448

      A sensitive monoclonal antibody (MAb) which simultaneously recognises aristolochic acids I and II was obtained. As one of the application of MAb 2A10-10B, an Eastern blotting method was newly developed for the visual-screening of AA-I and AA-II in Aristolochia and Asarum species. The established Eastern blotting method was also applied to the immunohistolocalisation of AA-I and AA-II in dry plant tissues.

    19. Rapid Quantitation of Furanocoumarins and Flavonoids in Grapefruit Juice using Ultra-Performance Liquid Chromatography (pages 654–660)

      Karen M. VanderMolen, Nadja B. Cech, Mary F. Paine and Nicholas H. Oberlies

      Version of Record online: 18 JUN 2013 | DOI: 10.1002/pca.2449

      Rapid (<5.0 min) ultra-performance liquid chromatography (UPLC) methods were developed for the determination of two furanocoumarins (bergamottin and 6',7'-dihydroxybergamottin) and four flavonoids (naringin, naringenin, narirutin, and hesperidin) in five grapefruit juice products. R2 values for the calibration curves of all analytes were >0.999. Considerable between-juice variation in the concentrations of these compounds was observed, and the quantities measured were in agreement with the concentrations published in HPLC studies.

    20. A LC/QTOF–MS/MS Application to Investigate Chemical Compositions in a Fraction with Protein Tyrosine Phosphatase 1B Inhibitory Activity from Rosa Rugosa Flowers (pages 661–670)

      Dongyu Gu, Yi Yang, Mahinur Bakri, Qibin Chen, Xuelei Xin and Haji Akber Aisa

      Version of Record online: 28 JUN 2013 | DOI: 10.1002/pca.2451

      As a traditional herbal medicine, Rosa rugosa flowers possess many activities. In this study, a LC-MS method was established for investigation of chemical compositions in a fraction with the PTP 1B inhibitory activity from Rosa rugosa. And 75 chemical compositions in this fraction were identified or tentatively characterized. The fragmentation pathways of gallotannins and ellagitannins were proposed. Knowing the phytochemical profiles of PTP 1B inhibitory activity fraction of this plant gives insights to its potential application to promote health.

    21. Elution-extrusion Counter-current Chromatography Separation of Two New Benzyl Ester Glucosides and Three Other High-polarity Compounds from the Tubers of Pleione bulbocodioides (pages 671–676)

      Yang Wang, Shu-hong Guan, Rui-hong Feng, Jing-xian Zhang, Qing Li, Xiao-hui Chen, Kai-shun Bi and De-an Guo

      Version of Record online: 24 JUN 2013 | DOI: 10.1002/pca.2453

      The tubers of Pleione bulbocodioides (Franch.) Rolfe, as a traditional Chinese medicine, have been used for the treatment of various cancers and bacterial infections. Elution-extrusion counter-current chromatography (EECCC), a support-free counter-current chromatography technique, showed its great advantage in separation of high-polarity compounds. In this research, we successfully used EECCC as a useful tool for the separation of gastrodin and benzyl ester glucosides, including two new ones from this herbal medicine.

    22. Analysis of Multiple Constituents in Cong-Ming-Tang, a Chinese Herbal Formula for the Treatment of Amnesia, by High-performance Liquid Chromatography with Quadrupole Time-of-flight Mass Spectrometry (pages 677–688)

      Yun Ling, Zhixiong Li, Mingcang Chen, Zhaolin Sun, Mingsong Fan and Chenggang Huang

      Version of Record online: 9 JUL 2013 | DOI: 10.1002/pca.2454

      This analytical tool allowed the identification of 55 compounds from CMT formulae by comparing their retentionylpropanoids. Onjisaponin MF was tentatively elucidated as a new triterpene saponin based on the summarised fragmentation rules. n times and MS spectra with those of authentic compounds or literature data in both positive and negative ion modes, including 4 xanthone C-glycosides, 4 sucrose esters, 11 oligosaccharide multi-esters,15 triterpene saponins, 15 triterpene acids, 2 lignans and 4 phe.