because of its low polarity. In both SFE and SFC quite often a
proportion of an organic solvent is added to the mobile phase as
modifier in order to widen the affinity of the mobile phase for the
different compounds. Moreover little variations in the density of
the fluid are achieved by small changes in pressure or temperature
which can further ameliorate the extraction or separation. Moreover, additional benefits in SFC compared to LC, are the use of a
much more ecological mobile phase with the reduction of organic
solvent utilization and costs.
Although recently, supercritical fluid chromatography (SFC)
coupled to mass spectrometry has gained attention as a green, fast
and useful technology applied to the carotenoid analysis [5–11],
and reports are available on the supercritical fluid extraction of
carotenoids [12–16], only three reports are available in the literature on the direct online extraction and determination of carotenoids and apocarotenoids by a supercritical fluid extraction–
supercritical fluid chromatography–mass spectrometry (SFE-SFCMS) methodology in food samples and biological fluids [17–
19]. The use of a triple-quadrupole mass spectrometer detector
(QqQ/MS) online with the SFE-SFC system is mandatory especially for the detection and identification of apocarotenoids. This is
caused by their usual very low abundance in food matrices (usually
in the order of 1–2% of the parent carotenoid) and the possibility to
perform multiple reaction monitoring (MRM) experiments which
greatly help in compounds identifications. Moreover, usually, in the
carotenoid field the mass spectrometer is operated using an atmospheric pressure chemical ionization (APCI) source with both the
positive and negative ionization modes. The purpose of this contribution was to provide an updated description of the SFE-SFCQqQ/MS methodology applied to the carotenoid and apocarotenoid analysis in various matrices, although with a particular focus
on apocarotenoid applications.
2 Materials
1. Carbon dioxide (CO 2 ), purity 99.9% or more.
2. Methanol (MeOH) and chloroform (CHCl 3 ), both LC-MS
grade.
3. 0.2 mL extraction vessel
4. Digital homogenizer.
2.1 Standards
1. Carotenoid commercial standards: β-carotene, zeaxanthin,
lutein,
β-cryptoxanthin,
capsanthin,
capsorubin,
β-apo-8
0 -carotenal, and physalein. Standards are prepared at a
concentration level of 10,000 mg L
À1 in a solution of 90%
MeOH and 10% CHCl 3 and stored in dark vials at À20
C.
210
Daniele Giuffrida et al.
proportion of an organic solvent is added to the mobile phase as
modifier in order to widen the affinity of the mobile phase for the
different compounds. Moreover little variations in the density of
the fluid are achieved by small changes in pressure or temperature
which can further ameliorate the extraction or separation. Moreover, additional benefits in SFC compared to LC, are the use of a
much more ecological mobile phase with the reduction of organic
solvent utilization and costs.
Although recently, supercritical fluid chromatography (SFC)
coupled to mass spectrometry has gained attention as a green, fast
and useful technology applied to the carotenoid analysis [5–11],
and reports are available on the supercritical fluid extraction of
carotenoids [12–16], only three reports are available in the literature on the direct online extraction and determination of carotenoids and apocarotenoids by a supercritical fluid extraction–
supercritical fluid chromatography–mass spectrometry (SFE-SFCMS) methodology in food samples and biological fluids [17–
19]. The use of a triple-quadrupole mass spectrometer detector
(QqQ/MS) online with the SFE-SFC system is mandatory especially for the detection and identification of apocarotenoids. This is
caused by their usual very low abundance in food matrices (usually
in the order of 1–2% of the parent carotenoid) and the possibility to
perform multiple reaction monitoring (MRM) experiments which
greatly help in compounds identifications. Moreover, usually, in the
carotenoid field the mass spectrometer is operated using an atmospheric pressure chemical ionization (APCI) source with both the
positive and negative ionization modes. The purpose of this contribution was to provide an updated description of the SFE-SFCQqQ/MS methodology applied to the carotenoid and apocarotenoid analysis in various matrices, although with a particular focus
on apocarotenoid applications.
2 Materials
1. Carbon dioxide (CO 2 ), purity 99.9% or more.
2. Methanol (MeOH) and chloroform (CHCl 3 ), both LC-MS
grade.
3. 0.2 mL extraction vessel
4. Digital homogenizer.
2.1 Standards
1. Carotenoid commercial standards: β-carotene, zeaxanthin,
lutein,
β-cryptoxanthin,
capsanthin,
capsorubin,
β-apo-8
0 -carotenal, and physalein. Standards are prepared at a
concentration level of 10,000 mg L
À1 in a solution of 90%
MeOH and 10% CHCl 3 and stored in dark vials at À20
C.
210
Daniele Giuffrida et al.