Methylation methods for the quantitative analysis of conjugated linoleic acid (CLA) isomers in various lipid samples.
Precise methylation methods for various chemical forms of conjugated linoleic acid (CLA), which minimize the formation of t,t isomers and allylmethoxy derivatives (AMD) with the completion of methylation, were developed using a 50 mg lipid sample, 3 mL of 1.0 N H(2)SO(4)/methanol, and/or 3 mL of 20% tetramethylguanidine (TMG)/methanol solution(s). Free CLA (FCLA) was methylated with 1.0 N H(2)SO(4)/methanol (55 degrees C, 5 min). CLA esterified in safflower oil (CLA-SO) was methylated with 20% TMG/methanol (100 degrees C, 5 min), whereas CLA esterified in phospholipid (CLA-PL) was methylated with 20% TMG/methanol (100 degrees C, 10 min), followed by an additional reaction with 1.0 N H(2)SO(4)/methanol (55 degrees C, 5 min). Similarly, CLA esterified in egg yolk lipid (CLA-EYL) was methylated by base hydrolysis, followed by reaction with 1.0 N H(2)SO(4)/methanol (55 degrees C, 5 min). These results suggest that for the quantitative analysis of CLA in lipid samples by GC, proper methylation methods should be chosen on the basis of the chemical forms of CLA in samples.
Park
Sook J
SJ
Division of Applied Life Sciences, and Institute of Agriculture and Life Sciences, Graduate School, Gyeongsang National University, Chinju 660-701, Korea.
Park
Cherl W
CW
Kim
Seck J
SJ
Kim
Jung K
JK
Kim
Young R
YR
Park
Kyung A
KA
Kim
Jeong O
JO
Ha
Yeong L
YL
D002849:Chromatography, Gas / Q000379:methods; D007202:Indicators and Reagents; D007536:Isomerism; D008041:Linoleic Acids / Q000032:analysis* / Q000737:chemistry; D008055:Lipids / Q000737:chemistry*; D000432:Methanol; D008745:Methylation; D010743:Phospholipids / Q000737:chemistry
D007202:Indicators and Reagents; D008041:Linoleic Acids; D008055:Lipids; D010743:Phospholipids; D000432:Methanol