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Updated: May 17, 2026
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Assaying for Inorganic Polyphosphate in Bacteria
Published on: January 21, 2019
Fluorometric quantification of polyphosphate in environmental plankton samples: extraction protocols, matrix effects,
Patrick Martin1, Benjamin A S Van Mooy
1Department of Marine Chemistry and Geochemistry, Woods Hole Oceanographic Institution, Woods Hole, Massachusetts, USA.
Applied and Environmental Microbiology
|October 30, 2012
Summary
Quantifying environmental polyphosphate (polyP) is challenging. A new method using boiling and proteinase K digestion, with DNase/RNase treatment, improves polyP measurement in marine samples.
Area of Science:
- Environmental science
- Biochemistry
- Analytical chemistry
Background:
- Polyphosphate (polyP) is a crucial biochemical and phosphorus pool.
- Methodological challenges hinder polyP quantification in environmental samples.
Purpose of the Study:
- To develop and validate a robust method for inorganic polyphosphate extraction and quantification in marine environments.
- To address challenges in polyP measurement, including matrix effects and nucleic acid interference.
Main Methods:
- Tested 15 protocols for polyP extraction from marine samples and cyanobacteria.
- Utilized fluorometric quantification with DAPI, including DNase and RNase treatments.
- Assessed matrix effects using standard addition and evaluated extraction efficiency.
Main Results:
- A protocol combining brief boiling and proteinase K digestion proved superior.
- Three successive extractions were necessary for complete polyP recovery.
- RNA interference was significant; DNase/RNase treatment reduced fluorescence by up to 57%.
- Matrix effects caused up to 50% over- or underestimation of polyP.
Conclusions:
- The optimized method enhances polyP quantification in marine particulate matter.
- Further validation is needed for diverse environmental sample types.
- Understanding matrix effects and nucleic acid interference is critical for accurate polyP measurements.
