Dataset Information

Methodology and validation of a new tandem mass spectrometer method for the quantification of inorganic and organic 18O-phosphate species.

ABSTRACT: Phosphorus (P) fertilizers are crucial to achieve peak productivity in agricultural systems. However, the fate of P fertilizers via microorganism incorporation and the exchange processes between soil pools is not well understood. 18Oxygen-labelled phosphate (18O- Pi) can be tracked as it cycles through soil systems. Our study describes biological and geochemical P dynamics using a tandem mass spectrometry (MS/MS) method for the absolute quantification of 18O- Pi. Soil microcosms underwent three treatments: (i) 18O- Pi, (ii) unlabelled phosphate (16O- Pi) or (iii) Milli-Q control, dissolved in a bio-stimulatory solution. During a 6-week series the microcosms were sampled to measure P by Hedley sequential fractionation and DNA extraction samples digested to 3'-deoxynucleoside 5'-monophosphates (dNMP). A MS/MS attached to a HPLC analyzed each P-species through collision-induced dissociation. The resin-extractable and bicarbonate 18O- Pi and 16O- Pi fractions displayed similar precipitation and adsorption-desorption trends. Biotic activity measured in the NaOH and dNMP fractions rapidly delabelled 18O- Pi; however, the MS/MS measured some 18O that remained between the P backbone and deoxyribose sugars. After 6 weeks, the 18O- Pi had not reached the HCl soil pool, highlighting the long-term nature of P movement. Our methodology improves on previous isotopic tracking methods as endogenous P does not dilute the system, unlike 32P techniques, and measured total P is not a ratio, dissimilar from natural abundance techniques. Measuring 18O- Pi using MS/MS provides information to enhance land sustainability and stewardship practices regardless of soil type by understanding both the inorganic movement of P fertilizers and the dynamic P pool in microbial DNA.

SUBMITTER: Schryer A

PROVIDER: S-EPMC7039501 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Methodology and validation of a new tandem mass spectrometer method for the quantification of inorganic and organic 18O-phosphate species.

Schryer Aimée A Bradshaw Kris K Siciliano Steven D SD

PloS one 20200224 2

Phosphorus (P) fertilizers are crucial to achieve peak productivity in agricultural systems. However, the fate of P fertilizers via microorganism incorporation and the exchange processes between soil pools is not well understood. 18Oxygen-labelled phosphate (18O- Pi) can be tracked as it cycles through soil systems. Our study describes biological and geochemical P dynamics using a tandem mass spectrometry (MS/MS) method for the absolute quantification of 18O- Pi. Soil microcosms underwent three ...[more]

PMID: 32092104

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Project description:There is an immediate need for improved methods to systematically and precisely quantify large sets of peptides in complex biological samples. To date protein quantification in biological samples has been routinely performed on triple quadrupole instruments operated in selected reaction monitoring mode (SRM), and two major challenges remain. Firstly, the number of peptides to be included in one survey experiment needs to be increased to routinely reach several hundreds, and secondly, the degree of selectivity should be improved so as to reliably discriminate the targeted analytes from background interferences. High resolution and accurate mass (HR/AM) analysis on the recently developed Q-Exactive mass spectrometer can potentially address these issues. This instrument presents a unique configuration: it is constituted of an orbitrap mass analyzer equipped with a quadrupole mass filter as the front-end for precursor ion mass selection. This configuration enables new quantitative methods based on HR/AM measurements, including targeted analysis in MS mode (single ion monitoring) and in MS/MS mode (parallel reaction monitoring). The ability of the quadrupole to select a restricted m/z range allows one to overcome the dynamic range limitations associated with trapping devices, and the MS/MS mode provides an additional stage of selectivity. When applied to targeted protein quantification in urine samples and benchmarked with the reference SRM technique, the quadrupole-orbitrap instrument exhibits similar or better performance in terms of selectivity, dynamic range, and sensitivity. This high performance is further enhanced by leveraging the multiplexing capability of the instrument to design novel acquisition methods and apply them to large targeted proteomic studies for the first time, as demonstrated on 770 tryptic yeast peptides analyzed in one 60-min experiment. The increased quality of quadrupole-orbitrap data has the potential to improve existing protein quantification methods in complex samples and address the pressing demand of systems biology or biomarker evaluation studies.

Dataset Information

Methodology and validation of a new tandem mass spectrometer method for the quantification of inorganic and organic 18O-phosphate species.

Publications

Methodology and validation of a new tandem mass spectrometer method for the quantification of inorganic and organic 18O-phosphate species.

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