Tailoring noise frequency spectrum between two consecutive second derivative filtering procedures to improve liquid chromatography-mass spectrometry determinations.

This paper reports a simple chemometric technique to alter the noise spectrum of a liquid chromatography-mass spectrometry (LC-MS) chromatogram between two consecutive second-derivative filter procedures to improve the peak signal-to-noise (S/N) ratio enhancement. This technique is to multiply one second-derivative filtered LC-MS chromatogram with another artificial chromatogram added with thermal noises prior to the other second-derivative filter. Because the second-derivative filter cannot eliminate frequency components within its own filter bandwidth, more efficient peak S/N ratio improvement cannot be accomplished using consecutive second-derivative filter procedures to process LC-MS chromatograms. In contrast, when the second-derivative filtered LC-MS chromatogram is conditioned with the multiplication alteration prior to the other second-derivative filter, much better ratio improvement is achieved. The noise frequency spectrum of the second-derivative filtered chromatogram, which originally contains frequency components within the filter bandwidth, is altered to span a broader range with multiplication operation. When the frequency range of this modified noise spectrum shifts toward the other regimes, the other second-derivative filter, working as a band-pass filter, is able to provide better filtering efficiency to obtain higher peak S/N ratios. Real LC-MS chromatograms, of which 5-fold peak S/N ratio improvement achieved with two consecutive second-derivative filters remains the same S/N ratio improvement using a one-step second-derivative filter, are improved to accomplish much better ratio enhancement, approximately 25-fold or higher when the noise frequency spectrum is modified between two matched filters. The linear standard curve using the filtered LC-MS signals is validated. The filtered LC-MS signals are also more reproducible. The more accurate determinations of very low-concentration samples (S/N ratio about 5-7) are obtained via standard addition procedures using the filtered signals rather than the determinations using the original signals.

Improving signal-to-noise ratios of liquid chromatography-tandem mass spectrometry peaks using noise frequency spectrum modification between two consecutive matched-filtering procedures.

This paper reports a simple chemometric technique to alter the noise spectrum of liquid chromatography-tandem mass spectrometry (LC-MS-MS) chromatogram between two consecutive matched filter procedures to improve the peak signal-to-noise (S/N) ratio enhancement. This technique is to multiply one match-filtered LC-MS-MS chromatogram with another artificial chromatogram added with thermal noises prior to the second matched filter. Because matched filter cannot eliminate low-frequency components inherent in the flicker noises of spike-like sharp peaks randomly riding on LC-MS-MS chromatograms, efficient peak S/N ratio improvement cannot be accomplished using one-step or consecutive matched filter procedures to process LC-MS-MS chromatograms. In contrast, when the match-filtered LC-MS-MS chromatogram is conditioned with the multiplication alteration prior to the second matched filter, much better efficient ratio improvement is achieved. The noise frequency spectrum of match-filtered chromatogram, which originally contains only low-frequency components, is altered to span a boarder range with multiplication operation. When the frequency range of this modified noise spectrum shifts toward higher frequency regime, the second matched filter, working as a low-pass filter, is able to provide better filtering efficiency to obtain higher peak S/N ratios. Real LC-MS-MS chromatograms containing random spike-like peaks, of which peak S/N ratio improvement is less than four times with two consecutive matched filters typically, are remedied to accomplish much better ratio enhancement approximately 16-folds when the noise frequency spectrum is modified between two matched filters.

Parametric studies of matched filters to enhance the signal-to-noise ratios of LC-MS-MS peaks.

Chromatographic parameters of reference signals employed in matched filter methods have been studied using numerical experiments to improve the signal-to-noise (S/N) ratios of small liquid chromatography (LC) peaks obtained with electrospray tandem mass spectrometers (MS-MS). These parameters include the width, shape, and S/N ratios of chromatographic peaks used as the reference signal profiles. Our results show the effect of reference peak widths on improving the S/N ratio of chromatographic peaks; the influence of reference peak shapes is negligible. To verify simulation results, various reference signals, including analyte peaks of high concentration standards, internal standard peaks, and artificial Gaussian peaks of different widths, have been employed to enhance signal peaks on real liquid chromatography-tandem mass spectrometry (LC-MS-MS) chromatograms via matched filter methods. Our experimental results demonstrate that the S/N ratio enhancement of chromatographic peaks agree with the simulation predictions. These findings, therefore, suggest that regardless of peak shape, a well-smooth peak with a width close to that of the analyte peak is an adequate reference signal, when matched filter methods are used to improve LC-MS-MS chromatograms. Nevertheless, all methods processed LC-MS-MS peaks in this study do not achieve the ideal improvement ratios estimated with simulation results. We attribute this deficiency to spike-like noise, which have considerable low frequency components riding on LC-MS-MS chromatograms. Matched filtering, which works as a low-pass filter in the frequency domain, cannot effectively eliminate low frequency flicker noise contributed by these spikes. In addition, simple median filtering does not provide adequate improvement despite being able to smooth out most spikes in the chromatograms.