A comparative study of methyl ester trimethylsilyl, allyldimethylsilyl and tert-butyldimethylsilyl ethers for electron impact mass spectrometry of leukotrienes.

The gas chromatographic and mass spectrometric properties of leukotriene B4, 20-hydroxy-leukotriene B4 and 20-carboxy-leukotriene-B4 were investigated as their methyl ester trimethylsilyl, methyl ester allyldimethylsilyl and methyl ester tert-butyldimethylsilyl ethers. The gas chromatographic properties of the trimethylsilyl and tert-butyldimethylsilyl ether derivatives were good with respect to peak shape and sensitivity, whereas the allyldimethylsilyl ether derivative gave a lower sensitivity. The sensitivity defined as the quantity that could be passed through the gas chromatographic column. The three derivatives showed a mass spectrometric fragmentation pattern with cleavage of the C12-C13 bond as an important feature. Particularly, the allyldimethylsilyl ether derivative of the three compounds studied exhibited a high tendency for C12-C13 bond cleavage resulting in a fairly intense ion at m/z 435. However, the mass spectra indicated multiple fragmentation pathways due to the presence of double bonds, leading to decreased intensities of the high mass ions. A quantitative analysis by selected ion monitoring of the most intense high mass ions in the respective mass spectrum demonstrated that neither derivative would allow measurements in the low picogram range. Catalytic hydrogenation of the double bonds was performed and the methyl ester trimethylsilyl, methyl ester allyldimethylsilyl and methyl ester tert-butyldimethylsilyl ether derivatives of the reduced compounds were prepared. Saturation of the double bonds increased the gas chromatographic sensitivity for the three derivatives as well as the intensities of the high mass ions in their mass spectra. The high sensitivity that can be obtained by measurement of such high mass ions was demonstrated by quantification of leukotriene B4 in lung tissue samples by selected ion monitoring.