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Molecules 2018, 23(8), 1851; https://doi.org/10.3390/molecules23081851

UHPLC Analysis of Saffron (Crocus sativus L.): Optimization of Separation Using Chemometrics and Detection of Minor Crocetin Esters

1
Dipartimento di Scienze Fisiche e Chimiche, Università degli Studi dell’Aquila, Via Vetoio, 67100 L’Aquila, Italy
2
Hortus Novus srl, Viale Aldo Moro 28 D, 67100 L’Aquila, Italy
*
Author to whom correspondence should be addressed.
Received: 29 June 2018 / Revised: 20 July 2018 / Accepted: 22 July 2018 / Published: 25 July 2018
(This article belongs to the Special Issue Qualitative and Quantitative Analysis of Bioactive Natural Products)
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Abstract

Ultra-high performance liquid chromatography (UHPLC) coupled with diode array detection (DAD) was applied to improve separation and detection of mono- and bis-glucosyl esters of crocetin (crocins), the main red-colored constituents of saffron (Crocus sativus L.), and other polar components. Response surface methodology (RSM) was used to optimise the chromatographic resolution on the Kinetex C18 (Phenomenex) column taking into account of the combined effect of the column temperature, the eluent flow rate and the slope of a linear eluent concentration gradient. A three-level full-factorial design of experiments was adopted to identify suitable combinations of the above factors. The influence of the separation conditions on the resolutions of 22 adjacent peaks was simultaneously modelled by a multi-layer artificial neural network (ANN) in which a bit string representation was used to identify the target analytes. The chromatogram collected under the optimal separation conditions revealed a higher number of crocetin esters than those already characterised by means of mass-spectrometry data and usually detected by HPLC. Ultra-high performance liquid chromatography analyses carried out on the novel Luna Omega Polar C18 (Phenomenex) column confirmed the large number of crocetin derivatives. Further work is in progress to acquire mass-spectrometry data and to clarify the chemical structure to the newly found saffron components. View Full-Text
Keywords: saffron; crocins; UHPLC analysis; separation optimisation; artificial neural network; response surface methodology saffron; crocins; UHPLC analysis; separation optimisation; artificial neural network; response surface methodology
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).
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D’Archivio, A.A.; Di Donato, F.; Foschi, M.; Maggi, M.A.; Ruggieri, F. UHPLC Analysis of Saffron (Crocus sativus L.): Optimization of Separation Using Chemometrics and Detection of Minor Crocetin Esters. Molecules 2018, 23, 1851.

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