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Anti-epileptic Contaminants

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The Severity of Anti-epileptic Contamination

Anti-epileptic is one of the best-selling drugs. They can eliminate or reduce seizures in two ways. Anti-epileptic drugs include phenytoin, carbamazepine, ethosuximide, sodium valproate, carbamazepine, ethosuximide, vanilla aldehyde, etc. Scientists have found that anti-epileptic drugs have become one of the most common drugs in surface water. Like most other drugs, it is excreted from the body in its natural form or as a metabolite and enters the water circulation. In addition, a large number of unused drugs were flushed down the toilet, causing the concentration of anti-epileptic drugs in the sewage to increase. Scientists found carbamazepine with a concentration as high as 3.8 μg/L in the influent of the wastewater treatment plant. However, in ordinary sludge treatment plants, the amount of carbamazepine removed is very small, or even negligible, which makes the environmental pollution of anti-epileptic more and more serious.

The Necessity of Monitoring Anti-epileptic Contamination with ELISA Testing

Fig. 1 The chemical structure of carbamazepine

After entering the environment, the degradation of anti-epileptic is very slow, with a half-life of about 100 days. Scientists have discovered that anti-epileptic in water pose a serious threat to aquatic life. For example, carbamazepine can change the blood characteristics of aquatic organisms (such as hemoglobin, total red blood cell count, etc.) and serum biomarkers (such as alkaline phosphate, lactate dehydrogenase, cholesterol, etc.). It also significantly affects oxidative stress markers (such as superoxide dismutase and catalase). To understand the anti-epileptic contaminants to the environment and achieve the purpose of controlling and treating the contaminants, as a fast and efficient immunoassay method, ELISA has long been used to monitor the anti-epileptic contaminants in environment.

ELISA Type

Direct competitive ELISA

The Advantages of ELISA Testing

Easy to use
Lower cost
High-throughput detection and analysis
High sensitivity and specificity
High accuracy and strong stability

ELISA Procedure for Anti-epileptic Contaminants Testing

Microtiter plates were coated with polyclonal antibody against mouse IgG (1 mg/L, 200 μL per well) in PBS. After incubating overnight at room temperature on a plate shaker at 750 rpm, the plates were washed three times with PBST.

Monoclonal antibody against carbamazepine diluted in PBS (8.6 μg/L, 200 μL per well) was added and incubated at room temperature for 1 h.

To establish the calibration curve, a CBZ stock solution was prepared in methanol and then further diluted with ultrapure water to obtain calibrators.

After washing the plate, pipette 25 μL of sample buffer into each well. After that, add standards and samples (175 μL per well). Then add the enzyme conjugate (50 μL per well), and shake the plate at room temperature for 30 min.

After washing the plate three times with PBST, substrate solution was added (200 μL per well) and incubated at room temperature for 30 min.

Terminated by the addition of 100 μL/well of 1M H₂SO₄, the absorbance was read at 450 nm and referenced to 650 nm.

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References

Zhang, Y.; et al. Carbamazepine and diclofenac: removal in wastewater treatment plants and occurrence in water bodies. Chemosphere. 2008,73(8): 1151-1161.
Scheytt, T.J.; et al. Mobility of pharmaceuticals carbamazepine, diclofenac, ibuprofen, and propyphenazone in miscible-displacement experiments. J Contam Hydrol. 2006,83(1-2): 53-69.
Bound, J.P.; et al. Household disposal of pharmaceuticals as a pathway for aquatic contamination in the United Kingdom. Environ Health Perspect. 2005,113(12): 1705-1711.
Bahlmann, A.; et al. Monitoring carbamazepine in surface and wastewaters by an immunoassay based on a monoclonal antibody. Anal Bioanal Chem. 2009,395(6): 1809-1820.

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