performing only static extraction with the empty vessel. In this
step the amount of co-solvent can be increased in order to
achieve higher extraction yields of the more polar compounds.
3. During this step two important aspects must be considered:
(1) the time of the dynamic extraction mode sufficient to flush
the entire extraction vessel (consider both time and flow rate),
and (2) the extracted analytes need to be trapped on the head of
the analytical column in order to achieve a good peak shape and
resolution. In this step a compromise must be found between
the amount of CO 2 and modifier.
4. The makeup pump flow is connected between the analytical
column and the BPR (as shown in Fig. 2). The aim of this
additional pump is dual: (1) to improve the ionization, especially when the electrospray ionization (ESI) interface is
employed, and (2) to improve the peak shape especially if a
low amount of organic modifier is employed. In fact, when a
low amount of organic solvent is employed the expansion of
the CO 2 after the BPR can cause the clogging of the transfer
tube to the MS. An additional trick in order to improve the
peak shape is to limit both the internal diameter of the transferring tube between the BPR and the MS system and the use of
unions than can cause an expansion of the CO 2 .
References
1. Rivera SM, Canela-Garayoa R (2012) Analytical tools for the analysis of carotenoids in
diverse materials. J Chromatogr A 1224:1–10
2. Amorim-Carrolho KT, Cepeda A, Fente C,
Regal P (2014) Review of methods for analysis
of carotenoids. Trends Anal Chem 56:49–73
3. Mercadante AZ, Rodrigues DB, Petri FC, Mariutti LRB (2017) Carotenoid esters in foods—a
review and practical directions on analysis and
occurrence. Food Res Int 99:830–850
4. Giuffrida D, Donato P, Dugo P, Mondello L
(2018) Recent analytical techniques advances
in the carotenoids and their derivative determination in various matrices. J Agric Food Chem
66:3302–3307
5. Jumaah F, Plaza M, Abrahamsson V, Turner C
(2016) A fast and sensitive method for the
separation of carotenoids using ultra-high performance supercritical fluid chromatographymass spectrometry. Anal Bioanal Chem
408:5883–5894
6. Li B, Zhao H, Liu J, Liu W, Fan S, Wu G, Zhao
GR (2015) Application of ultra-high performance supercritical fluid chromatography for
the determination of carotenoids in dietary
supplements. J Chromatogr A 1425:287–292
7. Berger TA, Berger BK (2013) Separation of
natural food pigments in saponified and
un-saponified paprika oleoresin by ultra-highperformance supercritical fluid chromatography
(UHPSFC).
Chromatographia
76:591–601
8. Matsubara A, Uchikata T, Shinohara M,
Nishiumi S, Yoshida M, Fukusaki E, Bamba T
(2012) Highly sensitive and rapid profiling
method for carotenoids and their epoxidized
products using supercritical fluid chromatography coupled with electrospray ionization-triple
quadrupole mass spectrometry. J Biosci Bioeng
113:782–787
9. Wada Y, Matsubara A, Uchikata T, Iwasaki Y,
Morimoto S, Kan K, Okura T, Fukusaki E,
Bamba T (2011) Metabolic profiling of
β-cryptoxanthin and its fatty acid esters by
supercritical fluid chromatography coupled
with triple quadrupole mass spectrometry. J
Sep Sci 34:3546–3552
10. Matsubara A, Bamba T, Ishida H, Fukusaki E,
Hirata K (2009) Highly sensitive and accurate
profiling of carotenoids by supercritical fluid
chromatography coupled with mass spectrometry. J Sep Sci 32:1459–1464
218
Daniele Giuffrida et al.
step the amount of co-solvent can be increased in order to
achieve higher extraction yields of the more polar compounds.
3. During this step two important aspects must be considered:
(1) the time of the dynamic extraction mode sufficient to flush
the entire extraction vessel (consider both time and flow rate),
and (2) the extracted analytes need to be trapped on the head of
the analytical column in order to achieve a good peak shape and
resolution. In this step a compromise must be found between
the amount of CO 2 and modifier.
4. The makeup pump flow is connected between the analytical
column and the BPR (as shown in Fig. 2). The aim of this
additional pump is dual: (1) to improve the ionization, especially when the electrospray ionization (ESI) interface is
employed, and (2) to improve the peak shape especially if a
low amount of organic modifier is employed. In fact, when a
low amount of organic solvent is employed the expansion of
the CO 2 after the BPR can cause the clogging of the transfer
tube to the MS. An additional trick in order to improve the
peak shape is to limit both the internal diameter of the transferring tube between the BPR and the MS system and the use of
unions than can cause an expansion of the CO 2 .
References
1. Rivera SM, Canela-Garayoa R (2012) Analytical tools for the analysis of carotenoids in
diverse materials. J Chromatogr A 1224:1–10
2. Amorim-Carrolho KT, Cepeda A, Fente C,
Regal P (2014) Review of methods for analysis
of carotenoids. Trends Anal Chem 56:49–73
3. Mercadante AZ, Rodrigues DB, Petri FC, Mariutti LRB (2017) Carotenoid esters in foods—a
review and practical directions on analysis and
occurrence. Food Res Int 99:830–850
4. Giuffrida D, Donato P, Dugo P, Mondello L
(2018) Recent analytical techniques advances
in the carotenoids and their derivative determination in various matrices. J Agric Food Chem
66:3302–3307
5. Jumaah F, Plaza M, Abrahamsson V, Turner C
(2016) A fast and sensitive method for the
separation of carotenoids using ultra-high performance supercritical fluid chromatographymass spectrometry. Anal Bioanal Chem
408:5883–5894
6. Li B, Zhao H, Liu J, Liu W, Fan S, Wu G, Zhao
GR (2015) Application of ultra-high performance supercritical fluid chromatography for
the determination of carotenoids in dietary
supplements. J Chromatogr A 1425:287–292
7. Berger TA, Berger BK (2013) Separation of
natural food pigments in saponified and
un-saponified paprika oleoresin by ultra-highperformance supercritical fluid chromatography
(UHPSFC).
Chromatographia
76:591–601
8. Matsubara A, Uchikata T, Shinohara M,
Nishiumi S, Yoshida M, Fukusaki E, Bamba T
(2012) Highly sensitive and rapid profiling
method for carotenoids and their epoxidized
products using supercritical fluid chromatography coupled with electrospray ionization-triple
quadrupole mass spectrometry. J Biosci Bioeng
113:782–787
9. Wada Y, Matsubara A, Uchikata T, Iwasaki Y,
Morimoto S, Kan K, Okura T, Fukusaki E,
Bamba T (2011) Metabolic profiling of
β-cryptoxanthin and its fatty acid esters by
supercritical fluid chromatography coupled
with triple quadrupole mass spectrometry. J
Sep Sci 34:3546–3552
10. Matsubara A, Bamba T, Ishida H, Fukusaki E,
Hirata K (2009) Highly sensitive and accurate
profiling of carotenoids by supercritical fluid
chromatography coupled with mass spectrometry. J Sep Sci 32:1459–1464
218
Daniele Giuffrida et al.