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How to Determine Aquatic Humic Substances

June 28th, 2010

By Lynda Huckestein, Project Chemist, Kelso, WA

How to Determine Aquatic Humic Substances

Aquatic Humic Substances (AHS) result from the decomposition of plant and animal residues and are found in soil, sediment, and water. They are widespread in nature and are mostly comprised from naturally occurring dissolved organic matter in water.

AHSs are an important component in water systems, particularly as they relate to the potential fate of chemicals in the aquatic environment. For example, upon chlorination, AHS may form trihalomethanes (THMs). THMs formation is a concern in drinking water systems and thus a determination for the presence of AHS is useful to evaluate potential sources of THMs. AHS can change the toxicity or the benefits of materials by making them more available or less available to organisms. These are just two examples of the impact AHS has on the fate and transport of analytes and environmental contaminates.

The procedure followed by Columbia Analytical for the determination of AHS is described in Standard Methods for the Examination of Water and Wastewater SM 5510B and SM 5310C. Samples collected for this procedure should be collected in organic free glass containers and should be stored in the dark at 4°C.

The amount of AHS present is determined by concentrating the water sample, at a pH of six, on an anion exchange column of diethylaminoethyl (DEAE) cellulose. The testing is performed in duplicate to evaluate precision in each matrix. The AHSs are removed from the DEAE cellulose under basic conditions and the eluent is acidified to minimize carbon dioxide contamination from the air. Any carbon dioxide that may be present is removed via purging with nitrogen. The concentration of dissolved organic carbon is then determined via persulfate oxidation and infrared spectroscopy. The AHS present is calculated as two times the dissolved organic carbon concentration.

Determining the level of AHS in an aquatic system provides information that may be useful in understanding the potential formation of biotoxic components and the impact to organisms present in the ecosystem.

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5 Responses to “How to Determine Aquatic Humic Substances”

  1. Mark Weisberg Says:

    What is the difference between the analysis for AHS vs. the analysis for Humic Acid?

  2. dilip .patil Says:

    Dear Sir,

    We want to know the method of Ammoniacal notrogen for determination and how to process it to remove from of effluent,


    Dilip Patil

  3. Dee O'Neill Says:

    Hello Dilip!

    There are a number of analytical methods to test for ammonia as nitrogen. You can view most of them at However, you must probably search a little more on-line for consultants that can give you options for removing nitrogen from your processes.

  4. Dee O'Neill Says:

    Hello Mark;

    Humic Acid is a specific component of AHS. The analysis for AHS is more general in nature. While humic acid may be a portion of AHS, the method does not allow for specific identification of humic acid in the sample.

  5. Barry Smith Says:

    Hi Dee, after spending a lot of the time using DEAE cellulose in the late 1980s extracting gram quantities of AHS from groundwater it is good to see its been developed as an analytical method for AHS. We also looked at using small DEAE columns and DEAE cellulose based SPE for establishing metal species bound to AHS. We also did some work showing that extra cellular polysaccharides also are exchanged by DEAE in addition to what more traditionally would be considered to be humic and fulvic acids / DOM.



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