LETTER TO THE EDITOR

Determination of Fatty Acids in Position 3 of Triglycerides

Sir: In the most frequently used method of
stereospecific triglyceride analysis, 1,2(2,3)-di-
27_p h
glycerides (obtained from triglycerides with + phospholipase A 2
pancreatic lipase or by Grignard deacylation)
are converted to l(3)-phosphatidylphenol, of - - Ph t-3
which only the sn-3-isomer is hydrolyzed by
phospholipase A 2 (Brockerhoff, H., Lipids pan-
6:942 [1971]). The fatty acids in positions 1 + 2 + creatic._ 2
and 2 can be directly analyzed, fatty acid 2 in
- Ph t-3 lipase +3
two different products, but the fatty acid
composition in 3 must be calculated by differ-
ence. Although two independent ways are open
for this calculation and the accuracy of the T h e conventional stereospecific analysis is fol-
analysis can be judged by the agreement of the lowed, but in an additional step the sn-l-phos-
results, a method to analyze the fatty acids in 3 pholipid, isolated by thin layer chromatog-
directly would still be desirable, not so much raphy, is hydrolyzed in position 3 by pancreatic
for additional confirmation of the analysis but lipase. Slotboom et al. (Chem. Phys. Lipids
because of the uncertainty involved in the 4:15 [ 197 0] ) have shown that this reaction can
quantitation of minor and trace fatty acids in 3 proceed to completion. Crude pancreas powder
merely by difference. A direct analysis of could be used, since the sn-l-phospholipid
position 3 is possible with a method starting would not be digested by pancreatic phospho-
from 1,3-diglyceride, but this intermediate can- lipase A 2. Esterification of the free phosphate
not be prepared from triglyceride except con- hydroxyl of the phospholipid, e.g., with diazo-
taminated with 6-10% isomerized methane, would accelerate the final lipolytic
1,2(2,3)-diglyceride (Brockerhoff, H., Lipids reaction.
6:942 [ 1971 ] ). A different method to analyze
position 3 directly is proposed in the following H. BROCKERHOFF
scheme (Q~) = phosphate; Ph = phenol). Fisheries Research Board of Canada
Halifax Laboratory
Halifax, Nova Scotia, Canada
2 lipase o r + 2 C6HsOPOC12
Grignard
3 3 H20 [ Received February 12, 1973]

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