Small molecules dominate organic phosphorus in NaOH-EDTA extracts of soils as determined by 31P NMR.

Understanding the composition of organic phosphorus (P) in soils is relevant to various disciplines, from agricultural sciences to ecology. Despite past efforts, the precise nature of soil organic P remains an enigma, especially that of the orthophosphate monoesters, which dominate 31P NMR spectra of NaOH-EDTA extracts of soils worldwide. The monoester region often exhibits an unidentified, broad background believed to represent high molecular weight (MW) P. We investigated this monoester background using 1D 31P NMR and 2D 1H31P NMR, as well as 31P transverse relaxation (T2) measurements to calculate its intrinsic linewidth and relate it to MW. Analyzing seven soils from different ecosystems, we observed linewidths of 0.5 to 3 Hz for resolved monoester signals and the background, indicating that it consists of many, possibly >100, sharp signals associated with small (<1.5 kDa) organic P molecules. This result was further supported by 2D 1H31P NMR spectra revealing signals not resolved in the 1D spectra. Our findings align with 31P NMR studies detecting background signals in soil-free samples and modern evidence that alkali-soluble soil organic matter consists of self-assemblies of small organic compounds mimicking large molecules.

Biomarkers for cheese authentication by detailed and fast gas chromatographic profiling of triacylglycerol fatty acids.

Unsaturated fatty acid isomers and odd- and branched-chain fatty acids (OBCFAs) in milk triacylglycerols (TAGs) can be quantitated using gas chromatography (GC), providing access to biomarkers of animal species, breeds, diet, geographic origin, and environmental conditions. Such analysis requires expensive cyanopropyl siloxane or ionic liquid columns of at least 50 m in length, which increases the elution time. Aiming to use GC for cheese authentication and characterization while keeping the experiment time short and maintaining a good separation between fatty acid (FA) isomers, we considered using a 30 m polyethylene glycol-2-nitroterephthalate column. The FAs thus quantitated allowed the discovery of specific biomarkers for the origins of cheese varieties highly consumed in several countries. In addition, the simple and multivariate correlations we found between FAs in the cheese TAG matrix were alternative means for characterization and authentication purposes.

Cheese characterization and authentication through lipid biomarkers obtained by high-resolution 1H NMR profiling.

Food quality and safety are at the heart of consumers' concerns across the world. Dairy products, because of their large consumption, are fertile ground for fraudulent acts. This fact justifies the development of effective, accessible, and rapid analytical methods for their authentication. A high-resolution spectral treatment method previously developed by our team was applied to 1H NMR spectra of cheese triacylglycerols. 178 Peaks were thus quantitated and successfully used in the construction of multivariate models for the quantitation of individual fatty acids and for the classification of cheese samples according to the producing species, to their origin and variety. Besides, several peaks related to the amount and position of anteisopentadecanoic, butyric, α-linolenic, myristoleic, rumenic, and vaccenic acids were, among others, specific biomarkers of cheese groups. For the first time in 1H NMR, we were able to identify and to quantitate signals related to minor fatty acids within cheese triacylglycerols.

High-resolution 1H NMR profiling of triacylglycerols as a tool for authentication of food from animal origin: Application to hen egg matrix.

Metabolomics of complex biological matrices conducted by means of 1H NMR leads to spectra suffering from severe signal overlapping. Previously, we have developed a high-resolution spectral treatment method to help solving this issue in 1H NMR of triacylglycerols. In this work, we tested the potential of the developed method in the characterization and authentication of food products from animal origin using egg yolk as a model matrix. The approach consisted in a spectral deconvolution guided by the precision obtained on the deconvoluted peaks after reference lineshape adjustment of spectra. Thus, 135 peaks were quantitated and successfully used as biomarkers of origin, of hens breed, and of farming system. This required multivariate statistical analyses for classification. The same pool of variables allowed construction of multivariate quantitation models for individual fatty acids. Furthermore, minute amounts of conjugated fatty acids were quantitated and used as fingerprints of samples from backyard and free-range farming.

A precise and rapid isotopomic analysis of small quantities of cholesterol at natural abundance by optimized 1H-13C 2D NMR.

Cholesterol, the principal zoosterol, is a key metabolite linked to several health complications. Studies have shown its potential as a metabolic biomarker for predicting various diseases and determining food origin. However, the existing INEPT (insensitive nuclei enhanced by polarization transfer) 13C position-specific isotope analysis method of cholesterol by NMR was not suitable for very precise analysis of small quantities due to its long acquisition time and therefore is restricted to products rich in cholesterol. In this work, a symmetric and adiabatic heteronuclear single quantum coherence (HSQC) 2D NMR sequence was developed for the high-precision (few permil) analysis of small quantities of cholesterol. Adiabatic pulses were incremented for improving precision and sensitivity. Moreover, several strategies such as the use of non-uniform sampling, linear prediction, and variable recycling time were optimized to reduce the acquisition time. The number of increments and spectral range were also adjusted. The method was developed on a system with a cryogenically cooled probe and was not tested on a room-temperature system. Our new approach allowed analyzing as low as 5 mg of cholesterol in 31 min with a long-term repeatability lower than 2‰ on the 24 non-quaternary carbon atoms of the molecule comparing to 16.2 h for the same quantity using the existing INEPT method. This result makes conceivable the isotope analysis of matrices low in cholesterol. Graphical abstract.