6. Integrate the chromatogram for peak areas, and calculate the
concentrations by using equations of corresponding calibration
curves (see Note 6) and by considering the weighed amount of
freeze-dried algal powder.
3.2.2 Fatty Acid Methyl
Esters by GC-MS
The GC-MS runs are controlled by MSD ChemStation 2.0 program (Agilent Technologies), and for identification the NIST Mass
Spectral Search Program (Version 2.0f) is used.
1. The oven temperature program is from 70
C (2 min) to
235
C at a rate of 10
C/min; total run time is 30 min.
2. The temperatures of the injector and MS source are 220 and
230
C, respectively.
3. The samples (2 μl) are injected with the Gerstel MPS injection
system.
4. The split ratio is 1:15–1:20.
5. The data are collected in electron impact (EI) mode (70 eV) at
a mass range of m/z 40–600.
6. Heptadecanoic acid methyl ester (C17:0 FAME) derived from
triheptadecanoate is used as internal standard for fatty acid
methyl esters.
3.2.3 Free Fatty Acids,
Sterols, and Other Polar
Compounds as TMS
Derivatives by GC-MS
Figure 2 shows a typical GC-MS total ion chromatogram (TIC)
obtained from algal lipid extract. In this case, the level of sterols and
squalene is very low. The hexadecatetraenoic acid (16:4n-3) is
known to appear in MGDG and DGDG.
1. The oven temperature program is from 70
C (1 min) to
270
C at a rate of 10
C/min; total run time is 30 min.
2. The temperatures of the injector and MS source are 220 and
230
C, respectively.
3. The samples (2 μl) are injected with the Gerstel MPS injection
system.
4. The split ratio is 1:15–1:20.
5. The data are collected in electron impact (EI) mode (70 eV) at
a mass range of m/z 40–600.
6. Trimethylsilyl ether of heptadecanoic acid (C17:0 FFA TMS) is
used as internal standard for trimethylsilyl derivatives (see Note 7).
4 Notes
1. Heptadecanoic acid (C17:0) as acyl group is used for quantification in UPLC-ELSD analyses of different lipid classes. For
calibration curves, five to nine concentration points
UPLC-ELSD Analysis of Algal Lipid Classes and Derivatization of Bound and. . .
229
concentrations by using equations of corresponding calibration
curves (see Note 6) and by considering the weighed amount of
freeze-dried algal powder.
3.2.2 Fatty Acid Methyl
Esters by GC-MS
The GC-MS runs are controlled by MSD ChemStation 2.0 program (Agilent Technologies), and for identification the NIST Mass
Spectral Search Program (Version 2.0f) is used.
1. The oven temperature program is from 70
C (2 min) to
235
C at a rate of 10
C/min; total run time is 30 min.
2. The temperatures of the injector and MS source are 220 and
230
C, respectively.
3. The samples (2 μl) are injected with the Gerstel MPS injection
system.
4. The split ratio is 1:15–1:20.
5. The data are collected in electron impact (EI) mode (70 eV) at
a mass range of m/z 40–600.
6. Heptadecanoic acid methyl ester (C17:0 FAME) derived from
triheptadecanoate is used as internal standard for fatty acid
methyl esters.
3.2.3 Free Fatty Acids,
Sterols, and Other Polar
Compounds as TMS
Derivatives by GC-MS
Figure 2 shows a typical GC-MS total ion chromatogram (TIC)
obtained from algal lipid extract. In this case, the level of sterols and
squalene is very low. The hexadecatetraenoic acid (16:4n-3) is
known to appear in MGDG and DGDG.
1. The oven temperature program is from 70
C (1 min) to
270
C at a rate of 10
C/min; total run time is 30 min.
2. The temperatures of the injector and MS source are 220 and
230
C, respectively.
3. The samples (2 μl) are injected with the Gerstel MPS injection
system.
4. The split ratio is 1:15–1:20.
5. The data are collected in electron impact (EI) mode (70 eV) at
a mass range of m/z 40–600.
6. Trimethylsilyl ether of heptadecanoic acid (C17:0 FFA TMS) is
used as internal standard for trimethylsilyl derivatives (see Note 7).
4 Notes
1. Heptadecanoic acid (C17:0) as acyl group is used for quantification in UPLC-ELSD analyses of different lipid classes. For
calibration curves, five to nine concentration points
UPLC-ELSD Analysis of Algal Lipid Classes and Derivatization of Bound and. . .
229