7 Analytical Protocols in Carotenoid Analysis
171
technique and its powerful capability in selectivity, limit of detection, and strategies applied for structural elucidation. However, the reader should consider that the
best LC-MS configuration for MS analysis of phytoplankton pigments may include
UHPLC-DAD system hyphenated with APCI source and Q-TOF mass analyzer, to
acquire MS spectra with full detailed information in different dimensions (MS with
mass accuracy and isotopic pattern of protonated ions, and subsequently tandem MS
with mass accuracy and isotopic pattern of product ions).
7.2.6.2 Nuclear Magnetic Resonance (NMR)
The progress in the hardware developments achieved in recent years has finally
allowed the hyphenation of LC with fine structural spectrometric analysis such as
1 H
NMR, which were technically banned for hyphenation. Now it is possible to build
metabolomic platforms that join the separation power of LC, the online acquisition
of UV-visible and MS spectra and the final structural confirmation by NMR of the
individual analytes annotated previously. Hence, the intense labor work of isolation
and purification of the individual compounds to proceed with the NMR spectrometry
has been released, and the high-throughput analysis providing definitive structural
identification of the analytes is a reality. However, still these advances are constrained
to large research facilities aimed to resolve metabolomic issues from different fields
than phytoplankton profiling. Several studies have been published dealing with NMR
of carotenoids in microalgae by using the classical routine of isolation and purification
of the target compound, and subsequent acquisition of the NMR spectra either in 1D
or 2D (Holtin et al. 2009; Sivathanu and Palaniswamy 2012). The development
of probes and the increase of the magnetic field have decreased the handicaps of
this technique, requiring less quantities of material and the option of performing
sequences on many nuclei. Even the option of acquiring NMR spectra directly from
tissue is available (Gaysinski et al. 2015) by means of the development of highresolution magic-angle spinning probes, a technique at the interface of the liquid
phase and the solid-state NMR, which is successfully applied to the analysis of
interest compounds in phytoplankton biomass (Simon et al. 2015).
Acknowledgements The financial support of the Ministerio de Economía y Competitividad
(MINECO-CICYT, Spanish Government, project AGL2017-87884-R) is gratefully acknowledged.
Antonio Pérez-Gálvez is a tenured scientist at the Spanish National Agency for Research (CSIC).
171
technique and its powerful capability in selectivity, limit of detection, and strategies applied for structural elucidation. However, the reader should consider that the
best LC-MS configuration for MS analysis of phytoplankton pigments may include
UHPLC-DAD system hyphenated with APCI source and Q-TOF mass analyzer, to
acquire MS spectra with full detailed information in different dimensions (MS with
mass accuracy and isotopic pattern of protonated ions, and subsequently tandem MS
with mass accuracy and isotopic pattern of product ions).
7.2.6.2 Nuclear Magnetic Resonance (NMR)
The progress in the hardware developments achieved in recent years has finally
allowed the hyphenation of LC with fine structural spectrometric analysis such as
1 H
NMR, which were technically banned for hyphenation. Now it is possible to build
metabolomic platforms that join the separation power of LC, the online acquisition
of UV-visible and MS spectra and the final structural confirmation by NMR of the
individual analytes annotated previously. Hence, the intense labor work of isolation
and purification of the individual compounds to proceed with the NMR spectrometry
has been released, and the high-throughput analysis providing definitive structural
identification of the analytes is a reality. However, still these advances are constrained
to large research facilities aimed to resolve metabolomic issues from different fields
than phytoplankton profiling. Several studies have been published dealing with NMR
of carotenoids in microalgae by using the classical routine of isolation and purification
of the target compound, and subsequent acquisition of the NMR spectra either in 1D
or 2D (Holtin et al. 2009; Sivathanu and Palaniswamy 2012). The development
of probes and the increase of the magnetic field have decreased the handicaps of
this technique, requiring less quantities of material and the option of performing
sequences on many nuclei. Even the option of acquiring NMR spectra directly from
tissue is available (Gaysinski et al. 2015) by means of the development of highresolution magic-angle spinning probes, a technique at the interface of the liquid
phase and the solid-state NMR, which is successfully applied to the analysis of
interest compounds in phytoplankton biomass (Simon et al. 2015).
Acknowledgements The financial support of the Ministerio de Economía y Competitividad
(MINECO-CICYT, Spanish Government, project AGL2017-87884-R) is gratefully acknowledged.
Antonio Pérez-Gálvez is a tenured scientist at the Spanish National Agency for Research (CSIC).