Quantification of selected furocoumarins by high-performance liquid chromatography and UV-detection: Capabilities and limits

► HPLC with UV detection is the usual method to quantify furocoumarins, with contradictory reported quantification limits (0.019–61mg/l). ► This collaborative test shows that this method is only applicable to simple cases, above a limit of 10mg/l. ► To be valid, each quantitative result should be su...

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Bibliographic Details
Published in:	Journal of Chromatography A Vol. 1257; pp. 34 - 40
Main Authors:	Macmaster, Angus P., Owen, Neil, Brussaux, Sylvain, Brevard, Hugues, Hiserodt, Richard, Leijs, Hans, Bast, Nikola, Weber, Berthold, Loesing, Gerd, Sherlock, Alan, Schippa, Christine, Vey, Matthias, Frérot, Eric, Tissot, Emeline, Chaintreau, Alain
Format:	Journal Article
Language:	English
Published:	Netherlands Elsevier B.V 28-09-2012
Subjects:	absorption Accuracy bias Chromatography Chromatography, High Pressure Liquid - methods Complex Mixtures - chemistry Confidence Confidence intervals Essential oils Furocoumarin Furocoumarins - analysis high performance liquid chromatography HPLC–UV Limit of quantification Mathematical analysis odors Oils, Volatile - chemistry psoralens Quantification Reproducibility of Results Sensitivity and Specificity Similarity Spectra Spectrophotometry, Ultraviolet - methods Validation Furocoumarin Validation Accuracy Quantification HPLC–UV Limit of quantification
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Summary:	► HPLC with UV detection is the usual method to quantify furocoumarins, with contradictory reported quantification limits (0.019–61mg/l). ► This collaborative test shows that this method is only applicable to simple cases, above a limit of 10mg/l. ► To be valid, each quantitative result should be supported by the positive identification of the analyte comparing its UV spectrum to that of a authentic substance. The performance of HPLC–UV as a means of quantifying selected furocoumarins in essential oils has been evaluated, based on a ring test validation approach. Accuracy profiles were generated, to determine bias and statistical confidence associated with determination at different concentrations, along with lower limits of quantification (LOQ). From these findings, it can be concluded that the method described may only be used in simple cases (essential oils), to measure individual furocoumarin compounds at concentrations greater than 10mg/l; the non compound-specific nature of detection by absorption in the UV range is unable to overcome the effect of interferences arising from chromatographic coelutions, such as those encountered in the analysis of complex commercial fragrance mixtures. The use of an algorithmically calculated ‘spectral similarity’ function, with reference to authentic standards, may be used to improve reliability in assignment and quantification.
Bibliography:	http://dx.doi.org/10.1016/j.chroma.2012.07.048 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0021-9673 1873-3778
DOI:	10.1016/j.chroma.2012.07.048