This document summarizes research evaluating the use of Supercritical Fluid Chromatography coupled with diode array detection (SFC-DAD) and mass spectrometry (SFC-MS) for the simultaneous detection of 23 fragrance allergens in cosmetic and scented products. Initial screening found the best results using a silica column and acetonitrile as the co-solvent. The SFC-UV method provided good resolution of the allergens in under 16 minutes, though further optimization is possible. Coupling SFC to a quadrupole time-of-flight mass spectrometer could help resolve any co-eluting peaks and allow detection of additional allergens.

1. The Cosmetic regulation (EC) N°1223/2009 regulates 26 fragrance allergens at the moment with an exceeding stipulated cut-off level. Among
them, two compounds are now partially forbidden (Evernia Prunastri, Evernia Furfuracea) in cosmetic and perfume products. Previous studies
using LC-UV has been already published1 leading to acceptable qualitative and quantitative results. Nevertheless acquisition time is quite long (> 30 min) and several co-
elutions can be observed.
In order to decrease analysis time and improve specificity, Supercritical Fluid Chromatography (SFC) coupled with diode array detector (DAD) has been tested. First step
consists in stationary phase screening (Si, C18, pentafluorophenyl (PFP), cyano (CN), Hypercarb) using isocratic mode (95% CO2 / 5% different co-solvent).
Best results have been obtained with Si column and Acetonitrile as co-solvent. 23 allergens of the actual list were analyzed during this study (others were not available in our
laboratories).
Thus we tried to improve the chromatographic condition to obtain the best separation of various constituents.
SFC-UV and SFC-MS methods for the simultaneous detection of 23
fragrance allergens in cosmetic and scented products
Cyrille Santerre1, Nadine Vallet1, David Touboul2
(1) Institut Supérieur International Parfum Cosmétique Arômes, Plateforme scientifique, ISIPCA, 34-36 rue du parc de Clagny, 78000 Versailles, France
(2) CNRS, Institut de Chimie des Substances Naturelles, UPR2301, Avenue de la Terrasse, 91190 Gif-sur-Yvette, France
INTRODUCTION
MATERIAL AND METHOD
RESULTS
CONCLUSION & OUTLOOK REFERENCES
[1] Villa C1, Gambaro R, Mariani E, Dorato S., High-performance liquid chromatographic method for
the simultaneous determination of 24 fragrance allergens to study scented products, J Pharm
Biomed Anal., 2007 Jul 27, 44(3), 755-62.
[2] Scientific Committee on Consumer Safety (SCCS)(page consultée le 22 janvier 2015), Document
PDF, [En ligne]. Adresse URL : ec.europa.eu/health/scientific_committees/consumer_safety/docs /
sccs_o_102.pdf
csanterre@isipca.fr, nvallet@isipca.fr, david.touboul@cnrs.fr
UV conditions :
Wavelength : 210 nm Width: 4 nm
Wavelength : 254 nm Width : 4 nm
Wavelength : 280 nm Width : 4 nm
Reference : 360 nm Width : 50 nm
Chromatographic conditions:
BPR pressure : 150 bar
Column temperature : 30 °C
Flow : 2,5 mL/min
Solvent : CO2 / ACN
Analysis duration : 16 minutes and 3
minutes post-run
Solution concentration 0,1% w/w
Injection volume : 1 µL
Column RXSIL 150mm*4,6mm*5µmAllergens studied
Name 210 nm 254 nm 280 nm Formula CAS Number
1 Alpha isométhyl ionone +++ ++ 0 C14H22O 127-51-5
2 Amyl cinnamal +++ +++ ++++ C14H18O 122-40-7
3 Anisyl alcohol ++ + + C8H10O2
105-13-5
4 Benzyl acohol ++ 0 0 C7H8O 100-51-6
5 Benzyl benzoate +++ + + C14H12O2
120-51-4
6 Benzyl cinnamate +++ +++ +++ C16H14O2
103-41-3
7 Benzyl salicylate +++ ++ + C14H12O3
118-58-1
8 Lilial ++ + 0 C14H20O 80-54-6
9 Cinnamal +++ +++ +++ C9H8O 104-55-2
10 Cinnamyl alcohol +++ +++ + C9H10O 104-54-1
11 Citral +++ ++ 0 C10H16O 5392-40-5
12 Citronellol ++ 0 0 C10H20O 106-22-9
13 Coumarine +++ ++++ +++ C9H6O2
91-64-5
14 Eugenol ++ 0 + C10H12O2
97-53-0
15 Farnesol ++ 0 0 C15H26O 4602-84-0
16 Geraniol ++ 0 0 C10H18O 106-24-1
17 Hexyl cinnamal +++ +++ +++ C15H20O 101-86-0
18 Hydroxycitronellal + 0 0 C10H20O2
107-75-5
19 Lyral ++ 0 0 C13H22O2
31906-04-4
20 isoeugenol ++ ++ ++ C10H12O2
97-54-1
21 Limonène ++ 0 0 C10H16
5989-27-5
22 Linalool ++ 0 0 C10H18O 78-70-6
23 Methyl heptin caronate ++ 0 0 C9H14O2
111-12-6
Time (Min) %A %B Flow (ml/min)
0,00 98,0 2,0 2,500
6,00 98,0 2,0 2,500
10,00 80,0 20,0 2,500
11,00 80,0 20,0 2,500
14,00 98,0 2,0 2,500
 First results using SFC-UV provide a correct resolution.
 Chromatographic parameters could be optimized.
 Coupling SFC-UV-QTOF could solve co-elution problem.
 Extend to new 82 cosmetic allergens2
Column C18 150mm*2,0mm*1,8µm
Column CN 100mm*2,1mm*1,8µm
Column HYPERCARB 100mm*2,1mm*5µm
Column PFP 150mm*2,0mm*3µm