The triglycerides of human plasma lipoproteins were separated with high performance reversed‐phase liquid chromatography. An octadecyl bonded 5‐μ silica column was used with a mobile phase of acetonitrile/acetone. Individual triglyceride types and critical pairs may be easily separated and identified.
The electron impact (EI) mass spectra of the permethyl, peracetyl and per(trifluoroacetyl) derivatives of hydroxylated benzo[a]pyrene (B[a]P) metabolites were determined and the fragmentation chemistry producing the spectra elucidated. The metabolites investigated were: 3-hydroxy-B[a]P; 7,8-dihydro-7,8-dihydroxy-B[a]P; 7,8,9,10-tetrahydro-7,8,9-trihydroxy-B[a]P; and 7,8,9,10-tetrahydro-7,8,9,10-tetrahydroxy-B[a]P. In addition, the positive and negative methane chemical ionization spectra were determined for the derivatives of the BP-tetrol. The EI fragmentation patterns of the methylated metabolites that contained partially saturated rings were complex and, in the case of the di- and trimethoxy compounds, included apparent violations of the even-electron rule. The permethylated triol and tetrol cleaved through a retro-Diels-Alder reaction. The EI spectra of the peracetates were dominated by losses of acetic acid and ketene. The per(trifluoroacetyl) species fragmented by losing elements of trifluoroacetic acid, trifluoracetate radical and trifluoroacetyl. The spectra obtained from the permethylated tetrol permitted accurate prediction of the corresponding permethylated derivatives of tetrol metabolites of chrysene and benz[a]anthracene. The ability to predict spectra may be useful in trace analysis of hydrocarbon metabolites in biological samples.
Cholesterol and cholesteryl esters were separated according to their carbon number and number of double bonds by high performance reversed-phase chromatography (HPRC) using acetonitrile/chloroform/methanol (1:1:1, v/v) as a mobile phase. It was found that within the same equivalent carbon number (ECN) category, cholesteryl esters with the highest number of double bonds eluted ahead of those with a lower number of double bonds, and with the cis isomers eluting ahead of their trans partners. Thus, cholesteryl oleate (C27-18:1c) elutes ahead of cholesteryl palmitate (C27-16:0) and ahead of cholesteryl elaidate (C27-18:1t). Human lipoprotein, as well as rat liver cholesteryl esters, were separated using this technique.
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