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Quantitative Analysis of Amygdalin and Prunasin in Prunus serotina Ehrh. using 1 H‐NMR Spectroscopy
Author(s) -
Santos Pimenta Lúcia P.,
Schilthuizen Menno,
Verpoorte Robert,
Choi Young Hae
Publication year - 2013
Publication title -
phytochemical analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.574
H-Index - 72
eISSN - 1099-1565
pISSN - 0958-0344
DOI - 10.1002/pca.2476
Subject(s) - chemistry , amygdalin , nuclear magnetic resonance spectroscopy , spectroscopy , analytical chemistry (journal) , chromatography , organic chemistry , medicine , alternative medicine , pathology , physics , quantum mechanics
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 CH 3 OH‐ d 4 and KH 2 PO 4 buffer in D 2 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‐ d 4 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. Conclusion 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 . Copyright © 2013 John Wiley & Sons, Ltd.