24. Avoid using injection volumes higher than 20 μL for preventing peak distortion during chromatography.
25. If xanthophyll esters are present in the sample and saponification has not been performed then the last step of the gradient
elution at 100% A should be increased to 10 min to allow
separation and detection of the very nonpolar diesters.
26. In general, 450 nm is a detection wavelength that can be used
for most carotenoids. However, for some particular carotenoids the detection wavelength needs to be adequate to their
chromophore properties. For instance, this is the case of phytoene and phytofluene with absorption maxima at 286 and
348 nm, respectively.
27. Calibration curves can be used for the estimation of other
carotenoids such as the case of cis isomers (or Z- isomers),
which are usually quantified using the calibration curve of the
trans (or all-E isomers) counterpart.
Acknowledgments
The author thanks to the Spanish Government (projects
AGL2017-85368P and CaRed Network BIO2015-71703-REDT
and BIO2017-90877-REDT) for financial support.
References
1. Britton G, Liaaen-Jensen S, Pfander H (2008)
Carotenoids handbook. Birkh€ auser Verlag,
Basel
2. Britton G, Hornero-Me ´ndez D (1997) Carotenoids and colour in fruit and vegetables. In:
Toma ´s-Barbera ´n FA, Robins RJ (eds) Phytochemistry of fruit and vegetables. Clarendon
Press, Oxford, pp 11–27
3. Rodriguez-Concepcion M, Avalos J, Bonet
ML, Boronat A, Gomez-Gomez L, HorneroMendez D, Limon MC, Mele ´ndez-Martı ´nez
AJ, Olmedilla-Alonso B, Palou A, Ribot J,
Rodrigo MJ, Zacarias L, Zhu C (2018) A
global perspective on carotenoids: metabolism,
biotechnology, and benefits for nutrition and
health. Prog Lipid Res 70:62–93. https://doi.
org/10.1016/j.plipres.2018.04.004
4. Britton G, Liaaen–Jensen S, Pfander H (2009)
Carotenoids, Nutrition and health, vol
5. Birkh€ auser Verlag, Basel, Switzerland
5. Ettre LS, Sakodynskii KI (1993) M. S. Tswett
and the discovery of chromatography I: early
work (1899–1903). Chromatographia 35
(3):223–231.
https://doi.org/10.1007/
BF02269707
6. Ettre LS, Sakodynskii KI (1993) M. S. Tswett
and the discovery of chromatography II: completion of the development of chromatography
(1903–1910).
Chromatographia
35
(5):329–338.
https://doi.org/10.1007/
BF02277520
7. Stewart I, Wheaton TA (1971) Continuous
flow separation of carotenoids by liquid chromatography. J Chromatogr 55:325–336
8. Khachik F (2009) Analysis of carotenoids in
nutritional
studies.
In:
Britton
G,
Liaaen–Jensen S, Pfander H (eds) Carotenoids,
vol 5: Nutrition and health, vol 5. Birkh€ auser
Verlag, Basel, Switzerland, pp 7-44
9. Mı ´nguez-Mosquera MI, Hornero-Me ´ndez D
(1993) Separation and quantification of the
carotenoid pigments in red peppers (Capsicun
annuum L.), paprika and oleoresin by
reversed-phase HPLC. J Agric Food Chem
41:1616–1620
10. Atienza SG, Ballesteros J, Martin A, HorneroMe ´ndez D (2007) Genetic variability of carotenoid concentration and degree of esterification
among
tritordeum
(xTritordeum
Ascherson et Graebner) and durum wheat
132
Da ´ maso Hornero-Me ´ ndez
25. If xanthophyll esters are present in the sample and saponification has not been performed then the last step of the gradient
elution at 100% A should be increased to 10 min to allow
separation and detection of the very nonpolar diesters.
26. In general, 450 nm is a detection wavelength that can be used
for most carotenoids. However, for some particular carotenoids the detection wavelength needs to be adequate to their
chromophore properties. For instance, this is the case of phytoene and phytofluene with absorption maxima at 286 and
348 nm, respectively.
27. Calibration curves can be used for the estimation of other
carotenoids such as the case of cis isomers (or Z- isomers),
which are usually quantified using the calibration curve of the
trans (or all-E isomers) counterpart.
Acknowledgments
The author thanks to the Spanish Government (projects
AGL2017-85368P and CaRed Network BIO2015-71703-REDT
and BIO2017-90877-REDT) for financial support.
References
1. Britton G, Liaaen-Jensen S, Pfander H (2008)
Carotenoids handbook. Birkh€ auser Verlag,
Basel
2. Britton G, Hornero-Me ´ndez D (1997) Carotenoids and colour in fruit and vegetables. In:
Toma ´s-Barbera ´n FA, Robins RJ (eds) Phytochemistry of fruit and vegetables. Clarendon
Press, Oxford, pp 11–27
3. Rodriguez-Concepcion M, Avalos J, Bonet
ML, Boronat A, Gomez-Gomez L, HorneroMendez D, Limon MC, Mele ´ndez-Martı ´nez
AJ, Olmedilla-Alonso B, Palou A, Ribot J,
Rodrigo MJ, Zacarias L, Zhu C (2018) A
global perspective on carotenoids: metabolism,
biotechnology, and benefits for nutrition and
health. Prog Lipid Res 70:62–93. https://doi.
org/10.1016/j.plipres.2018.04.004
4. Britton G, Liaaen–Jensen S, Pfander H (2009)
Carotenoids, Nutrition and health, vol
5. Birkh€ auser Verlag, Basel, Switzerland
5. Ettre LS, Sakodynskii KI (1993) M. S. Tswett
and the discovery of chromatography I: early
work (1899–1903). Chromatographia 35
(3):223–231.
https://doi.org/10.1007/
BF02269707
6. Ettre LS, Sakodynskii KI (1993) M. S. Tswett
and the discovery of chromatography II: completion of the development of chromatography
(1903–1910).
Chromatographia
35
(5):329–338.
https://doi.org/10.1007/
BF02277520
7. Stewart I, Wheaton TA (1971) Continuous
flow separation of carotenoids by liquid chromatography. J Chromatogr 55:325–336
8. Khachik F (2009) Analysis of carotenoids in
nutritional
studies.
In:
Britton
G,
Liaaen–Jensen S, Pfander H (eds) Carotenoids,
vol 5: Nutrition and health, vol 5. Birkh€ auser
Verlag, Basel, Switzerland, pp 7-44
9. Mı ´nguez-Mosquera MI, Hornero-Me ´ndez D
(1993) Separation and quantification of the
carotenoid pigments in red peppers (Capsicun
annuum L.), paprika and oleoresin by
reversed-phase HPLC. J Agric Food Chem
41:1616–1620
10. Atienza SG, Ballesteros J, Martin A, HorneroMe ´ndez D (2007) Genetic variability of carotenoid concentration and degree of esterification
among
tritordeum
(xTritordeum
Ascherson et Graebner) and durum wheat
132
Da ´ maso Hornero-Me ´ ndez