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A rapid method for developing chlorophyll equations using alternative solvents: DMF (N,N-dimethylformamide)

  • Methodology
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Journal of Applied Phycology Aims and scope Submit manuscript

Abstract

Acetone although very good as a spectroscopic solvent is often found to be an unsatisfactory extractant of chlorophylls from photosynthetic organisms and hence often seriously underestimates chlorophyll content. Unfortunately, although spectroscopic equations for organisms with chlorophyll a + b are available in some other solvents, suitable equations for organism with different chlorophyll profiles are often not available and are very labour-intensive and need a lot of expertise to obtain. DMF (N,N-dimethylformamide) is a very good extractant of chlorophylls. DMF is more toxic than DMSO (dimethyl sulphoxide) but it offers the same advantages of very low volatility and limited flammability but has a much lower freezing point. We describe a simple method to obtain usable chlorophyll equations in DMF solvent using available Acetone-based formulae as benchmarks. The workings for the development of the equations is shown so the same appoach can be used for other solvents. The relative errors of the equations are all about ± 2% (see Table).

  

Absorption coefficients (mg L−1 cm−1) in DMF solvent

Pigment Composition

Chlorophyll

630 nm

647 nm

664 nm

691 nm

Chl a only

Chl a

  

11.12 (± 0.150)

 

Chl a + b

Chl a

 

–2.223 (± 0.0523)

12.42 (± 0.270)

 
 

Chl b

 

23.45 (± 0.552)

–5.73 (± 0.125)

 

Chl a + c2

Chl a

–0.657 (± 0.00921)

 

11.47 (± 0.1188)

 
 

Chl c2

27.80 (± 0.390)

 

–3.64 (± 0.0377)

 

Chl a + c1c2

Chl a

-0.538 (± 0.0168)

 

10.85 (± 0.159)

 
 

Chl c1 + c2

32.77 (± 1.021)

 

-3.53 (± 0.0516)

 

Chl d + a

Chl d

  

–0.1296 (± 0.00191)

11.74 (± 0.1694)

 

Chl a

  

11.43 (± 0.1654)

–1.731 (± 0.0255)

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Data availability

The graphs of the spectrophotometric data used for the project are included in the Supplementary Material. The data as EXCEL files are available upon request.

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Acknowledgements

The authors wish to thank The Faculty of Technology and Environments for access to their facilities for this project.

Funding

No specific funding was used for the project.

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Authors and Affiliations

  1. Techology and Environment, Prince of Songkla University – Phuket, Phuket, Thailand

    Raymond J. Ritchie & Suhaliar Sma-Air

Authors
  1. Raymond J. Ritchie
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  2. Suhaliar Sma-Air
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Contributions

RJR conceived the project, RJR & SSA did the experimental work together and wrote the manuscript and prepared the figures. Both authors reviewed the manuscript and checked the revised MS and figures.

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Correspondence to Raymond J. Ritchie.

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Ritchie, R.J., Sma-Air, S. A rapid method for developing chlorophyll equations using alternative solvents: DMF (N,N-dimethylformamide). J Appl Phycol 36, 3153–3161 (2024). https://doi.org/10.1007/s10811-024-03358-z

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