Summary
While most other related OECD guidelines (e.g., 301, 302, 303, and 310) for aqueous aerobic biodegradability tests are only for freshwater environments, OECD 306 was developed due to the growing awareness of the need to protect the marine environment. Although the methods are similar to some of the OECD 301 methods in terms of system setup and parameter analysis, OECD 306 is not a test for ready biodegradability since no inoculum is added in addition to the microorganisms already present in the seawater. Neither do the tests simulate the marine environment since nutrients (mineral medium) are added and the concentration of test substance is much higher than would be present in the sea. There are two specified methods: Shake Flask Method and Closed Bottle Method. The Shake Flask Method allows for a maximum of 60 days, while the Closed Bottle method suggests a maximum of 28 days.
OECD 306 Testing Method
This standard contains two testing methods for examining the potential of marine biodegradation:
- The Shake Flask Method measures the dissolved organic carbon (DOC) removal during aerobic biodegradation in a shaking flask setup. The degradation percentage is calculated as the ratio of DOC removal to the initial DOC dosed into the test medium. 70% dissolved organic carbon removal indicates that there is a potential for biodegradation of the sample in the marine environment.
- The Closed Bottle Method measures the dissolved oxygen (DO) consumption during aerobic biodegradation in a closed bottle setup without head space. The degradation percentage is calculated as the ratio of DO consumption to the theoretical oxygen demand (ThOD). If the ThOD cannot be obtained, the chemical oxygen demand (COD) may be experimentally determined and used as a less satisfactory alternative. 60% dissolved oxygen consumption indicates that there is a potential for biodegradation of the sample in the marine environment.
The Shake Flask Method is capable of testing samples that are highly soluble, non-volatile, and non-adsorbing; while, the Closed Bottle Method is capable of testing samples that are highly soluble, volatile, and adsorbing.
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