title: HPLC analysis of oxindole alkaloids in Uncaria tomentosa: sample preparation and analysis optimisation by factorial design.
abstract: BACKGROUND
  Uncaria tomentosa ("cat's claw") is widely used for the treatment of some infectious and inflammatory diseases. Oxindole alkaloids are regarded as the most important components responsible for the biological activities attributed to the plant. Their analysis require efficient sample preparation and suitable reference standards but few are commercially available.
  
  
  OBJECTIVE
  To develop and validate a HPLC analytical method for oxindole alkaloids in Uncaria tomentosa with emphasis on sample preparation.
  
  
  METHODS
  Factorial experimental designs were used for the optimisation of both sample preparation and chromatographic separation. The optimised sample preparation involved extraction with aqueous ethanol, and the granulometry of the powdered plant material significantly influenced extraction yields. Mitraphylline was used as a calibration reference for the determination of total alkaloids.
  
  
  RESULTS
  The method was fully validated and showed good selectivity, linearity (r²  ≥ 0.9996), accuracy (≥ 96%) and precision (RSD < 2.4%). Detection and quantification limits for mitraphylline were 0.8 and 2.4 ppm, respectively.
  
  
  CONCLUSIONS
  The optimised chromatographic method, using organic buffer in the mobile phase, provided baseline separation of tetracyclic and pentacyclic alkaloids in the samples. Calibration using mitraphylline provided more accurate estimates of total alkaloid content when compared to other available reference alkaloids.
authors.affiliation: Departamento de Química, CP19081, Universidade Federal do Paraná, CEP 81531-990, Curitiba-PR, Brazil.
authors.forename: Gustavo
authors.initials: G
authors.lastname: Bertol
authors.forename: Luzia
authors.initials: L
authors.lastname: Franco
authors.forename: Brás Heleno de
authors.initials: BH
authors.lastname: Oliveira
mesh_terms: D000470:Alkaloids / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D002021:Buffers; D002138:Calibration; D020948:Cat's Claw / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D000431:Ethanol / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D026121:Indole Alkaloids / Q000032:analysis / Q000737:chemistry; D007211:Indoles / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D015394:Molecular Structure; D000078183:Oxindoles; D015203:Reproducibility of Results; D013696:Temperature; D013997:Time Factors; D014867:Water / Q000737:chemistry
chemical_list: D000470:Alkaloids; D002021:Buffers; D026121:Indole Alkaloids; D007211:Indoles; D000078183:Oxindoles; D014867:Water; C022960:2-oxindole; C509328:mitraphylline; D000431:Ethanol