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DEVELOPMENT AND VALIDATION OF AN HPLC-DAD METHOD TO ESTIMATE TOTAL KNOWN AND UNKNOWN FUROCOUMARIN CONTENT
EP25670
Poster Title: DEVELOPMENT AND VALIDATION OF AN HPLC-DAD METHOD TO ESTIMATE TOTAL KNOWN AND UNKNOWN FUROCOUMARIN CONTENT
Submitted on 03 Apr 2017
Author(s): Nicholas Housel (1), Ivan Domicio da Silva Souza (2), Rocco Burgo (1)
Affiliations: 1. INOLEX Inc.; 2. Rowan University
This poster was presented at PittCon 2017, Chicago, USA
Poster Views: 2,236
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Poster Information
Abstract: Furocoumarins are a family of natural chemicals with phototoxic potential, commonly found in plant extracts and fragrances. The European Cosmetics Directive 76/768/EEC limits the total content of furocoumarins in cosmetics to 1 ppm. Current methods define “total content” based on a certain number of external standards (markers), overlooking other unknown furocoumarins. An accurate total content analysis is unpractical, but we aimed to develop a method to estimate total content of both known and unknown furocoumarins. As a family of chemicals, different furocoumarins have similar UV absorption spectrum (see Fig. 1). Therefore, we propose an HPLC method based on external markers and the use of DAD-UV to identify unknown furocoumarins. Eight markers were studied: Bergapten; Bergamottin; Imperatorin; Isoimperatorin; Isopimpinellin; Oxypeucedanin; Xanthotoxin and 8-Geranyloxypsoralen. Elution gradient comprised water, acetonitrile and tetrahydrofuran through a C18 column. We obtained well resolved peaks (purity factors all above 999.9). Linearity proved to be satisfactory (R2= 0.9977 to 0.9998) in the range of 0.1 µg/mL to 2.5 µg/mL). Intra-day precision was highly noticed (RSD= 0.0865% to 1.73%) and inter-day precision was further confirmed (RSD= 0.0539% to 3.92%), except for Bergamottin and 8-Geranyloxypsoralen. Quantification limits ranged from 345.2 pg to 3421.4 pg. Percent recovery was determined using a plant extract not known to contain furocoumarins, and results were acceptable (85.68% to 114.03%), except for Bergamottin and 8-Geranyloxypsoralen. Robustness was studied by changing flow rate, oven temperature and concentration of mobile phase. The method was fully validated for 6 markers. DAD-UV detector allows us to identify unknown furocoumarins in between the markers and estimate their content based on linear regression of equivalent markers.Summary: Furocoumarins are commonly found in plant juices, extracts and oils, such as orange, lime, lemon, parsley, celery etc. These natural substances can cause burns to our skin if exposed to sunlight. Therefore, to ensure safety, the European Comission establishes a maximum limit of 1 ppm of furocoumarins in cosmetic formulation containing such extracts (EC Directive 76/768/EEC). This work aims to find a suitable solution for total furocoumarin determination.References: Frérot, E.; Decorzant, E. Quantification of Total Furocoumarins in Citrus Oils by HPLC Coupled
with UV, Fluorescence, and Mass Detection. J. Agric. Food Chem. 2004, 52, 6879–6886.
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