7 Analytical Protocols in Carotenoid Analysis
159
Table 7.1
Selected chromatographic methods applied for the analysis of phytoplankton pigments
Sample type
Targeted
pigments for
analysis
Stationary phase
Mobile phase
Detection/Identification References
Unialgal cultures
Carotenoids
and
chlorophylls
C
18 -5
μm, 250
× 4.6 mm
Gradient; A: 80:20 methanol: 0.5 M
aqueous ammonium acetate, pH 7.2
(v/v); B: 90:10 acetonitrile:water (v/v);
1 mL/min
UV/Vis: 436 nm
Wright et al.
(1991)
Marine phytoplankton
Carotenoids
and
chlorophylls
C
8 -3.5
μm, 150
× 4.6 mm
Gradient; A: 50:25:25
methanol:acetonitrile:0.25 M aqueous
pyridine solution (v/v/v); B:
20:60:20methanol:acetonitrile:acetone
(v/v/v); 1 mL/min
UV/Vis: 350–720 nm
Zapata et al.
(2000)
Microbial mat and
Chlorobium
phaeobacterioides
culture
Carotenoids
and
chlorophylls
Two in-line
SpherisorbODS2-C
18 -3
μm
colums,150
× 4.6 mm
Gradient; A: acetonitrile; B: methanol;
C: 0.01 M aqueous ammonium acetate;
D: ethyl acetate; 0.7 mL/min
UV/Vis:
300–800 nm—coupled
to ion trap and APCI a
source
Airs et al.
(2001, 2005)
Microalgae and
cyanobacteria samples
Carotenoids
and
chlorophylls
C
18 -3.5
μm, 150
× 4.6 mm
Gradient; A: 70:30 methanol:28 mM
aqueous ammonium acetate (v/v); B:
methanol; 0.8 mL/min
UV/Vis:
450 nm—coupled to
Q-ion trap and APCI
source/ESI interface
Frassanito
et al. (2005,
2008)
Natural phytoplankton
populations and
microalgal cultures
Carotenoids
and
chlorophylls
C
18 -5
μm, 150
× 3.9 mm
Gradient; A: 85:15 methanol:0.5 M
aqueous ammonium acetate (v/v); B:
90:10 acetonitrile:water (v/v); C: ethyl
acetate; 0.8 mL/min
UV/Vis: 400–750 nm
van Leeuwe
et al. (2006)
Chlorella pyrenoidosa
Carotenoids C
30 -5
μm, 250
× 4.6 mm
Gradient; A: 84:14:2
methanol:acetonitrile:water (v/v/v); B:
methylene chloride; 1 mL/min
UV/Vis: 450 nm
Inbaraj et al.
(2006)
(continued)