A convenient method for determination of the C20-22 PUFA composition of glycerolipids in blood and breast milk.
A convenient method was developed for preparation of FAME in small amounts from glycerolipids of blood or breast milk. Initially, 0.04-0.06 mL blood or breast milk was spotted onto a small piece of filter paper (1.5 x 1.5 cm) that had been washed with acetone containing 0.05% 2,6-di-tert-butyl-p-cresol (BHT). Each piece, once it had dried, was put in a small test tube, to which 2 mL hexane and 0.2 mL 2 M KOH/methanol were added. After vigorous mixing or sonication for 2 min at room temperature, the solution was neutralized or acidified by the addition of a few drops of acetic acid. To the solution was added 2 mL H2O, and then the hexane layer that separated was concentrated to dryness in vacuo. The FAME obtained were analyzed by GC. The method was applicable to the analysis of a large number of blood and breast milk samples, and the arachidonate/(eicosapentaenoate + docosahexaenoate) ratios could be determined rapidly.
Ichihar
Kenichi
K
Biological Chemistry, Faculty of Agriculture, Kyoto Prefectural University, Shimogamo, Japan. ichihara@kpu.ac.jp
Waku
Kanako
K
Yamaguchi
Chiaki
C
Saito
Kazumi
K
Shibahara
Akira
A
Miyatani
Shuichi
S
Yamamoto
Kohei
K
D002849:Chromatography, Gas / Q000379:methods; D005231:Fatty Acids, Unsaturated / Q000032:analysis* / Q000097:blood*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008895:Milk, Human / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013010:Sonication
D005231:Fatty Acids, Unsaturated