By David W. Russell
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Example text
This mass spectrometer has the added benefit of providing structural information when operated in the MS-MS mode that can be compared with putative synthetic standards of the candidate ligands. Electron-Impact Mass Spectrometry Electron-impact (EI) fragmentation spectra are highly reproducible and predictive with specific functional groups and substructures yielding distinctive fragmentation patterns. Therefore, the ability to generate such spectra on the target compounds can be highly beneficial.
Samples dissolved in as little as 10 l can be used with conical autosampler vials. GC analysis is particularly well suited for the analysis of small volatile lipophilic molecules, but does have important limitations with respect to certain types of natural product characterizations. Thermal instability and excessive fragmentation may prevent collection of useful spectra. , those with more than one hydroxyl group, may be difficult to volatize. Carboxylic acids are refractory generally to analysis unless derivatized.
The standard curve is obtained by analyzing mixtures of varying amounts of unlabeled 4b-hydroxycholesterol with a constant amount of [2H6]4bhydroxycholesterol, preferably the same amount that was added to the biological sample. It is not necessary to run the standards through the chromatography step before analysis; however, it is advisable to make a new standard curve for each batch of analyses. Typically the standards are run prior to the samples, and both the plotting of the curve and the subsequent analysis of samples is done automatically by the chromatography software.
