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Preparation and analysis of non-polar metabolites
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Nonpolar metabolites were hydrolyzed
and trimethylsilylated.
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Hydrolysis was performed with 0.5 mL
of 1.25 M HCl in methanol and 0.8 mL chloroform at 50°C
for five hours.
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Following hydrolysis, the solvents
and HCl were evaporated under nitrogen.
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The sample was then resuspended in
80mL
pyridine and 20mL
MSTFA + 1% TMCS and incubated for 1hr at 50°C.
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One mL
of the sample was injected at 1:1 split rationas described for polar
samples, with the exception that the oven temperature was held at 315°C
for 11 minutes.
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