Lipids
Plasma lipoproteins play an important role in the metabolism of lipids. This is an analysis using a GFC column,OHpak SB-805 HQ by mixed modes of GFC and reversed phase.
Sample : Lipoprotein (Sigma)
LDL
HDL
VLDL
Columns : Shodex OHpak SB-805 HQ (8.0mmID*300mm) x 2 Eluent : 50mM Tris + 0.3M NaCl Flow rate : 0.5mL/min Detector : UV(280nm) Column temp. : Room temp.
This shows an analysis using an affinity column, AFpak ADS-894, bonded with dextran sulfonate as a ligand. The column is suitable for the fractionation and purification of plasma lipoproteins.
Sample : Lipoprotein (Sigma)
LDL
HDL
VLDL
Column : Shodex AFpak ADS-894 (8.0mmID*50mm)
Eluent : (A); 0.05M Sodium phosphate buffer(pH7.4)
(B); (A) + 1.0M NaCl
Flow rate : 1.0mL/min
Detector : UV(280nm)
Column temp. : Room temp.
This shows an analysis performed using a multisolvent GPC column ( Asahipak GS-310 7G ) with acetone as an eluent.
GS-310 7G is an old type of GF-310 HQ.
Sample :
Triolein
Diolein
Monoolein
Oleic acid
Column : Shodex Asahipak GS-310 7G (7.5mmID*500mm) Eluent : Acetone Flow rate : 1.0mL/min Detector : RI Column temp. : 25deg-C
T. Nishio et al., Agric. Biol. Chem. 52(5) (1988) 1203
Fatty Acids were analyzed using RSpak DS-613 (a column for reversed phase chromatography).
Sample :
1. Caprylic acid, CH3(CH2)6COOH
2. Myristic acid, CH3(CH2)12COOH
3. Linoleic acid, C17H31COOH
4. Palmitic acid, CH3(CH2)14COOH
5. Stearic acid, CH3(CH2)16COOH
Column : Shodex RSpak DS-613 (6.0mmID*150mm) Eluent : H2O/CH3CN=7/95 Flow rate : 0.8mL/min Detector : Shodex RI Column temp. : 40deg-C
Fatty acids were analyzed using ODSpak F-411 (a column for reversed phase chromatography).
Sample :
1. EPA, Eicosapentaenoic acid)
2. DHA, Docosahexaenoic acid)
3. Arachidonic acid
4. Linoleic acid
Column : Shodex ODSpak F-411 (4.6mmID*150mm) Eluent : H2O/CH3CN=20/80 Flow rate : 0.7mL/min Detector : Shodex RI Column temp. : 30deg-C
Fatty acids were analyzed using RSpak DE-413 (a column for reversed phase chromatography).
Sample : CnH2n+1COOH, 40micro-L
1. Capric acid (n=9)
2. Lauric acid (n=11)
3. Myristic acid (n=13)
4. Palmitic acid (n=15)
5. Stearic acid (n=17)
6. Arachidic acid, Eicosanoic acid (n=19)
7. Behenic acid, Docosanoic acid (n=21)
Column : Shodex RSpak DE-413 (4.6mmID*150mm) Eluent : CH3CN/THF/H2O=45/20/35 Flow rate : 0.5mL/min Detector : Shodex RI Column temp. : 40deg-C
Fatty acids were analyzed using RSpak RP18-413 (a column for reversed phase chromatography). Capric acid could be eluted after V0 with addition of 0.1% H3PO4 to eluent.
Sample : CnH2n+1COOH, 30micro-L
1. Capric acid (n=9)
2. Lauric acid (n=11)
3. Myristic acid (n=13)
4. Palmitic acid (n=15)
5. Stearic acid (n=17)
6. Arachidic acid, Eicosanoic acid (n=19)
Column : Shodex RSpak RP18-413 (4.6mmID*150mm) Eluent : 0.1% H3PO4 aq./CH3CN/THF=30/50/20 Flow rate : 0.7mL/min Detector : Shodex RI Column temp. : 40deg-C
Fatty acids were analyzed using GPC KF-402HQ ((a semi-micro organic SEC (GPC) column)).
Sample : CnH2n+1COOH, 10micro-L
1. Melissic acid (n=29) 0.1%
2. Cerotic aid (n=25) 0.1%
3. Behenic acid (n=21) 0.1%
4. Arachidic acid (n=19) 0.1%
5. Stearic acid (n=17) 0.1%
6. Palmitic acid (n=15) 0.1%
7. Myristic acid (n=13) 0.1%
8. Lauric acid (n=11)
9. n-Decanoic acid (n=9) 0.1%
10. n-Octanoic acid (n=7) 0.1%
11. n-Hexanoic acid (n=5) 0.3%
12. n-Valeric acid (n=4) 0.3%
Columns : Shodex GPC KF-402HQ (4.6mmID*250mm) x 2 Eluent : THF Flow rate : 0.3mL/min Detetor : Shodex RI(small cell volume) Column temp. : 40deg-C
Fatty acid methyl esters were analyzed using a polymer-based reversed-phase column RSpak DS-413.
Sample : 0.2% each, 20micro-L
1. Methyl linoleate
2. Methyl palmitate
3. Methyl oleate
4. Methyl stearate
Column : Shodex RSpak DS-413 (4.6mmID*150mm) Eluent : CH3CN/THF/H2O=45/30/25 Flow rate : 1.0mL/min Detetor : Shodex RI Column temp. : 40deg-C
LTs (Leukotrienes) were analyzed using Asahipak ODP-50 6D ( a column for reversed phase chromatography ).
Sample : Leukotrienes
Column : Shodex Asahipak ODP-50 6D (6.0mmID*150mm) Eluent : 0.2M CH3COONa buffer(pH4.5 adjusted with CH3COOH)/CH3CN/CH3OH=50/25/25 Flow rate : 1.0mL/min Detector : UV(280nm) Column temp. : 30deg-C
Courtesy of Dr. Oishi and Dr. Yamaki, Kitazato University
LTs produced by polymorphonuclear leukocyte of rat were analyzed using Asahipak ODP-50 6D ( a column for reversed phase chromatography ).
Sample : Leukotrienes
Column : Shodex Asahipak ODP-50 6D (6.0mmID*150mm) Eluent : 0.2M CH3COONa buffer(pH4.5 adjusted with CH3COOH)/CH3CN/CH3OH=50/25/25 Flow rate : 1.0mL/min Detector : UV(280nm) Column temp. : 30deg-C
Courtesy of Dr. Oishi and Dr. Yamaki, Kitazato University
HETEs (hydroxyeicosatetraenoic acids) were analyzed using Asahipak ODP-50 6D ( a column for reversed phase chromatography ).
Sample : HETE
1. 15-HETE
2. 12-HETE
3. 5-HETE
Column : Shodex Asahipak ODP-50 6D (6.0mmID*150mm) Eluent : H2O/CH3COOH/CH3CN/CH3OH=50/0.1/50/10 Flow rate : 1.0mL/min Detector : UV(235nm) Column temp. : 30deg-C
Courtesy of Dr. Oishi and Dr. Yamaki, Kitazato University
HEETs produced by polymorphonuclear leukocyte of rat were analyzed using Asahipak ODP-50 6D ( a column for reversed phase chromatography ).
Sample : 5-HETE
Column : Shodex Asahipak ODP-50 6D (6.0mmID*150mm) Eluent : H2O/CH3COOH/CH3CN/CH3OH=50/0.1/50/10 Flow rate : 1.0mL/min Detector : UV(235nm) Column temp. : 30deg-C
Courtesy of Dr. Oishi and Dr. Yamaki, Kitazato University