Sediment Phosphorus Extractants for Phosphorus‐31 Nuclear Magnetic Resonance Analysis

Ahlgren, Joakim; Brabandere, Heidi De; Reitzel, Kasper; Rydin, Emil; Gogoll, Adolf; Waldebäck, Monica

doi:10.2134/jeq2006.0235

Cited by 44 publications

(28 citation statements)

References 37 publications

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“…Phosphorus-31 nuclear magnetic resonance spectroscopy requires large sample sizes and may be insensitive to polyP tightly associated with metal cations in solid granules. Moreover, alkaline extractions may incompletely extract and partly hydrolyze polyP (30,31). Assays based on enzymes of polyP metabolism (either exopolyphosphatase followed by orthophosphate [PO 4 3Ϫ ] determination [32,33] or polyP kinase followed by ATP measurement [34,35]) entail purifying polyP, but purification procedures suffer from low yields (36,37) and there is no commercial source for the necessary enzymes.…”

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confidence: 99%

Fluorometric Quantification of Polyphosphate in Environmental Plankton Samples: Extraction Protocols, Matrix Effects, and Nucleic Acid Interference

Martin

Mooy

2013

Appl Environ Microbiol

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Polyphosphate (polyP) is a ubiquitous biochemical with many cellular functions and comprises an important environmental phosphorus pool. However, methodological challenges have hampered routine quantification of polyP in environmental samples. We tested 15 protocols to extract inorganic polyphosphate from natural marine samples and cultured cyanobacteria for fluorometric quantification with 4=,6-diamidino-2-phenylindole (DAPI) without prior purification. A combination of brief boiling and digestion with proteinase K was superior to all other protocols, including other enzymatic digestions and neutral or alkaline leaches. However, three successive extractions were required to extract all polyP. Standard addition revealed matrix effects that differed between sample types, causing polyP to be over-or underestimated by up to 50% in the samples tested here. Although previous studies judged that the presence of DNA would not complicate fluorometric quantification of polyP with DAPI, we show that RNA can cause significant interference at the wavelengths used to measure polyP. Importantly, treating samples with DNase and RNase before proteinase K digestion reduced fluorescence by up to 57%. We measured particulate polyP along a North Pacific coastal-to-open ocean transect and show that particulate polyP concentrations increased toward the open ocean. While our final method is optimized for marine particulate matter, different environmental sample types may need to be assessed for matrix effects, extraction efficiency, and nucleic acid interference. Inorganic polyphosphate (polyP) is a linear polymer of three to several hundred phosphate residues that appears to be found in cells of all organisms (1). It is required for many cellular functions, including bacterial virulence (2-4), biofilm formation (4, 5), quorum sensing (4), competitive fitness in the environment (6), microbial stress responses (3,7,8), and survival during stationary phase (7, 9-11). PolyP can also act as a phosphorus (P) store in microbes. PolyP breakdown allows cells to survive during P limitation (12, 13), although some microbes transiently accumulate polyP upon nitrogen and P stress (8,14,15). Massive accumulation of polyP by certain bacteria sequesters dissolved P during biological wastewater treatment (16)(17)(18).While polyP is studied in a range of microbiological and environmental fields, the lack of easy quantitative methods has hampered progress. For example, polyP has very rarely been measured in the oceans despite increasing interest in the physiological responses of marine microbes to the ultralow P concentrations frequently found at sea and their biogeochemical implications (14,(19)(20)(21)(22)(23)(24)(25). PolyP has been found at nanomolar concentrations, comprising around 10% of total particulate P (26-28). PolyP also plays a role in redox cycling and geological sequestration of P (26, 27) and potentially in phytoplankton iron storage (29).Common analytical approaches have significant drawbacks. Phosphorus-31 nuclear magnetic resonance ...

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Fluorometric Quantification of Polyphosphate in Environmental Plankton Samples: Extraction Protocols, Matrix Effects, and Nucleic Acid Interference

Martin

Mooy

2013

Appl Environ Microbiol

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“…To reduce interference of paramagnetic ions (Fe and Mn) in 31 P NMR measurements, 5% (v/v) bicarbonate-buffered dithionite (BD) solution (0.11 M NaHCO 3 + 0.11 M Na 2 S 2 O 4 ) was added to the concentrated extracts (Zheng et al, 2013). The BD solution can reduce Fe and Mn to states more ready to release adsorbed P compounds (Ahlgren et al, 2007). The extracts were then freeze-dried and re-dissolved in 0.5 mL of D 2 O and 0.2 mL of M NaOH.…”

Section: P Nmrmentioning

confidence: 99%

Pyrolysis temperature affects phosphorus transformation in biochar: Chemical fractionation and 31P NMR analysis

Zhang

Shao

et al. 2016

Science of The Total Environment

127

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Phosphorus (P) recycling or reuse by pyrolyzing crop residue has recently elicited increased research interest. However, the effects of feedstock and pyrolysis conditions on P species have not been fully understood. Such knowledge is important in identifying the agronomic and environmental uses of biochar. Residues of three main Chinese agricultural crops and the biochars (produced at 300°C-600°C) derived from these crops were used to determine P transformations during pyrolysis. Hedley sequential fractionation and 31 P NMR analyses were used in the investigation. Our results showed that P transformation in biochar was significantly affected by pyrolysis temperature regardless of feedstock (Wheat straw, maize straw and peanut husk). Pyrolysis treatment transformed water soluble P into a labile (NaHCO 3 -P i ) or semi-labile pool (NaOH-P i ) and into a stable pool (Dil. HCl P and residual-P). At the same time, organic P was transformed into inorganic P fractions which was identified by the rapid decomposition of organic P detected with solution 31 P NMR. The P transformation during pyrolysis process suggested more stable P was formed at a higher pyrolysis temperature. This result was also evidenced by the presence of less soluble or stable P species, such as such as poly-P, crandallite (CaAl 3 Contents lists available at ScienceDirectScience of the Total Environment j o u r n a l h o m e p a g e : w w w . e l s e v i e r . c o m / l o c a t e / s c i t o t e n v P forms at higher pyrolysis temperature although their solubility or stability requires further investigation. Results suggested that a relatively lower pyrolysis temperature retains P availability regardless of feedstock during pyrolysis process.

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“…Cade-Menun and Preston [5] found that an aqueous solution containing both NaOH and EDTA extracted the greatest diversity of P forms and the largest percentage of total phosphorus in soil. Recently, it has been shown that a 0.25 M NaOH and 0.05 M EDTA extracting solution was most efficient in solubilizing a maximum range of organic P forms from sediments [26,28], while minimizing the co-extraction of Fe (III) and Mn (II) metals, which may significantly affect the resolution and intensity of 31 P NMR spectra [14,1]. Addition of HCl or Chelex to extracts was also found to be useful in reducing interferences [13,24].…”

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confidence: 99%

Optimized procedure for the determination of P species in soil by liquid-state 31P-NMR spectroscopy

Mazzei

Cozzolino

et al. 2015

Chem. Biol. Technol. Agric.

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Sediment Phosphorus Extractants for Phosphorus‐31 Nuclear Magnetic Resonance Analysis

Cited by 44 publications

References 37 publications

Fluorometric Quantification of Polyphosphate in Environmental Plankton Samples: Extraction Protocols, Matrix Effects, and Nucleic Acid Interference

Fluorometric Quantification of Polyphosphate in Environmental Plankton Samples: Extraction Protocols, Matrix Effects, and Nucleic Acid Interference

Pyrolysis temperature affects phosphorus transformation in biochar: Chemical fractionation and 31P NMR analysis

Optimized procedure for the determination of P species in soil by liquid-state 31P-NMR spectroscopy

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