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Authentication of herbal medicines and nutraceuticals is growing in importance, especially as the global economy grows and products are shipped worldwide. A useful approach is chemical profiling or spectral fingerprinting of plant extracts [1, 2, 3, 4, 5]. Although less sensitive than mass spectrometry (MS), nuclear magnetic resonance (NMR) spectroscopy provides a more reproducible complementary technique for the identification and quantification of metabolites in plant extracts [6].
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NMR is a key method for metabolomics and the number of papers has been growing exponentially as demonstrated by a nice review [7]. However, much of this growth has been in targeted analysis for which sets of metabolites are identified and quantified in the NMR spectrum. For authentication and screening, especially in industry, a faster and easier untargeted analysis approach is provided by chemical profiling which is also known as spectral fingerprinting. These approaches avoid the inherent problems in selecting and quantifying peaks in complex NMR spectra. Chemical profiling is an untargeted analysis for which the individual components of the botanical material are not identified or quantified; instead, the spectra are compared point by point using chemometric classifiers. The use of NMR for untargeted profiling coupled to chemometrics is a burgeoning and important application area. Here are some nice reviews on the topic of NMR metabolic profiling [8, 9, 10, 11, 12].
Typically for NMR spectroscopy the real spectral component of the Fourier transform of the free induction decay (FID) is used. After phase correction, the real absorbance spectrum has sharp and symmetric peaks. However, additional information in the imaginary dispersion spectrum is only used for phase-correcting the real spectrum. Because the rotating magnetization vector is modeled in the complex plane by using only the real spectrum some of the analytical signal is unused. The use of the magnitude or amplitude spectrum is proposed because this spectrum although less visually appealing will have greater signal-to-noise and reproducibility compared to the real absorbance spectrum. Reproducibility is important for classification or pattern recognition approaches to work effectively. The increase of signal in the magnitude spectrum results from the greater peak areas of the wider peaks than those found in the real spectrum. This finding is not surprising because it is a trading rule between signal and resolution [13].
NMR was used to profile four diverse sets of extracts. The samples were classified using six different classification methods. The average classification rates were statistically compared between the real NMR absorbance spectra and the magnitude spectra obtained from the absolute value of the complex spectrum. All the validations used 100 bootstrapped Latin partitions (BLPs) [14].
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