title: Determination of terpenoid indole alkaloids in hairy roots of Rhazya stricta (Apocynaceae) by GC-MS.
abstract: BACKGROUND
  Rhazya stricta Decne. (Apocynaceae) is a medicinal plant rich in terpenoid indole alkaloids (TIAs), some of which possess important pharmacological properties. The study material including transgenic hairy root cultures have been developed and their potential for alkaloid production are being investigated.
  
  
  OBJECTIVE
  In this study, a comprehensive GC-MS method for qualitative and quantitative analysis of alkaloids from Rhazya hairy roots was developed.
  
  
  METHODS
  The composition of alkaloids was determined by using GC-MS. In quantification, the ratio between alkaloid and internal standard was based on extracted ion from total ion current (TIC) analyses.
  
  
  RESULTS
  The developed method was validated. An acceptable precision with RSD ≤ 8% over a linear range of 1 to 100 µg/mL was achieved. The accuracy of the method was within 94-107%. Analysis of hairy root extracts indicated the occurrence of a total of 20 TIAs. Six of them, pleiocarpamine, fluorocarpamine, vincamine, ajmalicine and two yohimbine isomers are reported here for the first time in Rhazya. Trimethylsilyl (TMS) derivatisation of the extracts resulted in the separation of two isomers for yohimbine and also for vallesiachotamine. Clearly improved chromatographic profiles of TMS-derivatives were observed for vincanine and for minor compounds vincamine and rhazine.
  
  
  CONCLUSIONS
  The results show that the present GC-MS method is reliable and well applicable for studying the variation of indole alkaloids in Rhazya samples.
authors.affiliation: VTT Technical Research Centre of Finland Ltd, P.O. Box 1000, Tietotie 2, 02044-VTT, Espoo, Finland.|Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, P.O. Box 56, FI-00014, Helsinki, Finland.
authors.forename: Amir
authors.initials: A
authors.lastname: Akhgari
authors.affiliation: Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, P.O. Box 56, FI-00014, Helsinki, Finland.
authors.forename: Into
authors.initials: I
authors.lastname: Laakso
authors.affiliation: VTT Technical Research Centre of Finland Ltd, P.O. Box 1000, Tietotie 2, 02044-VTT, Espoo, Finland.
authors.forename: Tuulikki
authors.initials: T
authors.lastname: Seppänen-Laakso
authors.affiliation: Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, P.O. Box 56, FI-00014, Helsinki, Finland.
authors.forename: Teijo
authors.initials: T
authors.lastname: Yrjönen
authors.affiliation: Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, P.O. Box 56, FI-00014, Helsinki, Finland.
authors.forename: Heikki
authors.initials: H
authors.lastname: Vuorela
authors.affiliation: VTT Technical Research Centre of Finland Ltd, P.O. Box 1000, Tietotie 2, 02044-VTT, Espoo, Finland.
authors.forename: Kirsi-Marja
authors.initials: KM
authors.lastname: Oksman-Caldentey
authors.affiliation: VTT Technical Research Centre of Finland Ltd, P.O. Box 1000, Tietotie 2, 02044-VTT, Espoo, Finland.
authors.forename: Heiko
authors.initials: H
authors.lastname: Rischer
mesh_terms: D000470:Alkaloids / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D029062:Apocynaceae / Q000737:chemistry* / Q000235:genetics; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D007536:Isomerism; D015394:Molecular Structure; D018517:Plant Roots / Q000737:chemistry* / Q000235:genetics; D030821:Plants, Genetically Modified; D015203:Reproducibility of Results; D046948:Secologanin Tryptamine Alkaloids / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D046509:Tissue Culture Techniques / Q000379:methods; D014297:Trimethylsilyl Compounds / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D014749:Vincamine / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D015016:Yohimbine / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D000470:Alkaloids; D046948:Secologanin Tryptamine Alkaloids; D014297:Trimethylsilyl Compounds; C114765:pleiocarpamine; D015016:Yohimbine; D014749:Vincamine
keywords: GC-MS; Rhazya stricta; extracted ion monitoring; terpenoid indole alkaloids; transformed hairy roots; trimethylsilyl derivatisation