Q1)

The toxicity process used was Phase I of TIE (Toxicity Identification Evaluation). This uses in vivo tests. It uses effluent (in this study: leachate
from landfill) at different concentration on test organisms (in this study: fish, prawns and tomato seeds). This method can also be used to
identify toxicants in sediments, ambient waters and other complex mixtures. The TIE method comprises three related phases; toxicants are
characterized in Phase I, identified in Phase II and their identity confirmed in Phase III. The TIE method comprises three related phases;
toxicants are characterized in Phase I, identified in Phase II and their identity confirmed in Phase III. With the approach of this method, the
number of constituents associated with the toxicants can be reduced before analyses began and some knowledge of physical/chemical
characteristics is gained. This approach simplifies the analytical problems and reduces cost.
Bioassay by using plant (phytotoxicity test) has the advantages of the seeds remaining viable on the shelf for a long time and of the test being
simple and not requiring major equipment.

Some of the things measured in this study were BOD, COD, DO, pH, TDS, TSS, conductivity, alkalinity, ammoniacal nitrogen,
nitrate\phosphate\sulphate concentrations. Other procedures performed were pH adjustments, filtration, aeration and C18 solid phase
extraction, EDTA chelation and oxidant reduction.
TIE gives fantastic data of ecological relevance, considers bioavailability and is good for potentially all toxic compounds. It is however limited in
its ability to identify the toxicant.

The other major toxicity process available is EDA (Effects-directed analysis), which uses in vitro endpoints. It emphasises organic contaminants
as the cause of observed toxicity. EDA uses biological effects to help narrow down the number of potential toxicants. With EDA, extracts are
performed right from the start (organic solvent, solid phases extraction, etc.) with a focus on organic compounds. (to the detriment of
potential non organic toxicants). Pre-concentrated and purified samples are used with the whole array of instrumentation. In vitro testing used
to narrow the range of possible compounds, which is cheaper and quicker.
Unlike TIE, EDA gives no specific consideration to bioavailability (what fraction of toxicant exists in a form that is able to be absorbed and
metabolised in an organism). EDA puts emphasis on organic compounds while TIE considers all potentially toxic chemicals. EDA uses organic
extracts from samples whereas TIE uses whole samples.

Relating to this study, if EDA was to be performed, organic compounds would be extracted from the leachate samples. We can ten check the
effect of the different fractions on mutagenicity, bioluminescence, algal growth and daphniid toxicity.

Q2)
Phase I (characterisation) of TIE was used because the researchers needed to characterize the toxicants of a Malaysian landfill leachate. They
didnt want to focus only on organic compounds and thus didnt choose EDA.

Q3)
OD was used to quantitatively determine the level of dissolved Oxygen in the samples.
pH was used to determine if the chemicals were mostly of basic or acidic nature. (in this case they were mostly basic)

Q4)
The Phase I tests indicated the presence of organic compounds. As part of Phase II, Chromatography could be performed and the eluted
substances tested individually for toxicity. There was high concentration of ammonia detected so as part of Phase II, the researchers could test
specifically for NH3.

Q5)
The results indicated that the leachate contained toxicants which are basic in nature, unstable under acidic pHs (possibly organic acids), some
oxidants as well as nonpolar organic compounds.

Q6)
Further test that are recommended are looking into various known biological outcomes of toxic materials (e.g. endocrine disruption, estrogen
activity, dioxin and dioxin-like compound activity, cytochrome P-450 activity, mutagenicity, bioluminescence, algal growth and daphnid
toxicity.