Spectral Library Assembly

The Dephy data-processing package offers several proprietary noise filtering options. We have not yet determined unequivocally whether any of these data treatments increase reproducibility or concentrate the taxonomic information content of the averaged spectra, although first results suggest that use of filtered data can improve spectral reproducibility and thus assist the ruggedness of sample classification. 

4 SPECTRAL LIBRARY ASSEMBLY 5.4.1 Causes of Spectral Drift 

Pyrolysis spectra become distorted with respect to their diagnostic features for two major sets of reasons. The first is variations in instrument operation (e.g., heat transfer efficiency from wire to sample, ion source temperature, MAB gas identity, analyzer calibration, tuning, and ion transmission discrimination attributable to contaminated optics). Most of these factors can be controlled 

Over a long time period it may well not be possible to duplicate library cell culture conditions. What happens when the lot of media used in the final culture step prior to pyrolysis has been consumed Can culture media suppliers assure nutritional identity between batches Media types for growth of fastidious strains invariably include natural products such as brewer s yeast, tryptic soy, serum, egg, chocolate, and/or sheep blood. Trace components in natural products cannot be controlled to assure an infinite, invariable supply. The microtiter plate wells used here do not hold much media. Even so, the day will come when all media supplies are consumed and a change in batch is unavoidable. When that happens, if there were no effective way to compensate spectra for the resulting distortions, it would be necessary to re-culture and re-analyze replicates for every strain in the reference library. Until recently the potential for obsolescence was a major disincentive for developing PyMS spectral libraries of bacteria. Why this is no longer an insurmountable problem is discussed in the next section. 

The major causes of spectral variation were (1) instrumental drift, as Goodacre and Kell realized, but also (2) sample history, as discussed above. In particular, variations in the supplier or even the batch of tryptic soy agar (TSA) used for cell culturing led to spectral variations that differed in degree among disparate species. This phenomenon was attributed to the differential metabolic capabilities of the species with respect to the changed nutrients. 

---

METHOD REPRODUCIBILITY AND SPECTRAL LIBRARY ASSEMBLY FOR RAPID BACTERIAL CHARACTERIZATION BY METASTABLE ATOM BOMBARDMENT PYROLYSIS MASS SPECTROMETRY... 

Electron ionization was the first ionization method developed for mass spectrometry, and it remains the most widely used. The most extensive mass spectral libraries assembled are those of electron ionization mass spectra recorded under a standard set of conditions. The ion-... 

Spectral drift compensation for initial and ongoing assembly as well as consultation of a PyMS spectral library. 

The El source has been the most widely used ion source over the past 60 years and continues to be the method of choice for the analysis (either qualitative or quantitative) of small- to medium-sized volatile organic compounds. The inherent reproducibility of the mass spectra has enabled the assembly of large spectral libraries. Computers associated with current generation instruments can efficiently (in a few seconds) search an unknown mass spectrum against tens of thousands of reference spectra in order to aid in the identification of an analyte. The general scheme of an El source includes the introduction of the vaporized analyte molecules into the ionization chamber, exposure of those molecules... 

Intelligent computer assisted interpretation of spectroscopic data should be based on the knowledge from large structure oriented data collections. Both the inspection of spectral features and the statistical evaluation of similar structures (from library searches) can provide a set of probability ranked substructures which are readily assembled to target structures. The idea of substructure analysis allows the chemist to combine the results of different interpretation strategies, different databases and different spectroscopic methods to yield the structural information desired. Thus in a multidimensional data system hke SPECINFO structural noise can be effectively suppressed, if all information available in the spectroscopic laboratory is combined in a central intelligent computer system. 

When applying infi-ared spectroscopy to the preliminary qualitative identification of polymers it is desirable to assemble a library of standard spectra. Razinger et al have described a minicomputer procedure for digitization, maintenance and retrieval of an infared spectral collection. 

---