title: Quantitative analysis of amygdalin and prunasin in Prunus serotina Ehrh. using (1) H-NMR spectroscopy.
abstract: BACKGROUND
  Prunus serotina is native to North America but has been invasively introduced in Europe since the seventeenth century. This plant contains cyanogenic glycosides that are believed to be related to its success as an invasive plant. For these compounds, chromatographic- or spectrometric-based (targeting on HCN hydrolysis) methods of analysis have been employed so far. However, the conventional methods require tedious preparation steps and a long measuring time.
  
  
  OBJECTIVE
  To develop a fast and simple method to quantify the cyanogenic glycosides, amygdalin and prunasin in dried Prunus serotina leaves without any pre-purification steps using (1) H-NMR spectroscopy.
  
  
  METHODS
  Extracts of Prunus serotina leaves using CH3 OH-d4 and KH2 PO4 buffer in D2 O (1:1) were quantitatively analysed for amygdalin and prunasin using (1) H-NMR spectroscopy. Different internal standards were evaluated for accuracy and stability. The purity of quantitated (1) H-NMR signals was evaluated using several two-dimensional NMR experiments.
  
  
  RESULTS
  Trimethylsilylpropionic acid sodium salt-d4 proved most suitable as the internal standard for quantitative (1) H-NMR analysis. Two-dimensional J-resolved NMR was shown to be a useful tool to confirm the structures and to check for possible signal overlapping with the target signals for the quantitation. Twenty-two samples of P. serotina were subsequently quantitatively analysed for the cyanogenic glycosides prunasin and amygdalin.
  
  
  CONCLUSIONS
  The NMR method offers a fast, high-throughput analysis of cyanogenic glycosides in dried leaves permitting simultaneous quantification and identification of prunasin and amygdalin in Prunus serotina.
authors.affiliation: Departamento de Química, Instituto de Ciências Exatas, Universidade Federal de Minas Gerais, Av. Antônio Carlos 6627, Belo Horizonte, MG, 31270-901, Brazil; Natural Products Laboratory, Institute of Biology Leiden, Leiden University, 2300 RA, Leiden, The Netherlands.
authors.forename: Lúcia P
authors.initials: LP
authors.lastname: Santos Pimenta
authors.forename: Menno
authors.initials: M
authors.lastname: Schilthuizen
authors.forename: Robert
authors.initials: R
authors.lastname: Verpoorte
authors.forename: Young Hae
authors.initials: YH
authors.lastname: Choi
mesh_terms: D000678:Amygdalin / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D006027:Glycosides / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D006859:Hydrogen / Q000032:analysis; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D009570:Nitriles / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D010936:Plant Extracts / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D018515:Plant Leaves / Q000737:chemistry; D011422:Propionates / Q000592:standards; D027861:Prunus / Q000737:chemistry*; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors; D014297:Trimethylsilyl Compounds / Q000592:standards
chemical_list: D006027:Glycosides; D009570:Nitriles; D010936:Plant Extracts; D011422:Propionates; D014297:Trimethylsilyl Compounds; C007173:cyanogenic glycosides; C019063:prunasin; D000678:Amygdalin; D006859:Hydrogen
keywords: Prunus serotina.; amygdalin; cyanogenic glycosides; prunasin; quantitative NMR