Malaysian Journal of Analytical Sciences Vol 21 No 3 (2017): 518 - 526

DOI: https://doi.org/10.17576/mjas-2017-2103-01

 

 

 

ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA)-BASED-SENSOR FOR DETERMINATION OF BENZO[A]PYRENE IN RIVER WATER USING SCREEN-PRINTED GOLD ELECTRODE

 

(Sensor-berasaskan Ujian Enzim-Pengerap Immuno (ELISA) untuk Pengesanan Benzo[a]pirina dalam Air Sungai Menggunakan Elektrod Emas Skrin Tercetak)

 

Noor Sheryna Jusoh, Alyza Azzura Abd Rahman Azmi, Azrilawani Ahmad*

 

School of Marine and Environmental Sciences,

Universiti Malaysia Terengganu, 21030 Kuala Terengganu, Terengganu, Malaysia

 

*Corresponding author: azrilawani.ahmad@umt.edu.my

 

 

 

Received: 16 August 2016; Accepted: 15 April 2017

 

 

Abstract

This paper described the surface modification of screen-printed gold electrode using thiol self-assembled monolayer, in combination with enzyme-linked immunosorbent assay (ELISA) format for determination of benzo[a]pyrene (BaP) in water sample. All biomolecules were immobilized on the surface of working electrode to increase the sensitivity of the developed biosensor by applying different types of ELISA. Electrochemical characterizations using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were carried out to study the conductivity behavior of the developed biosensor. Based on the results, biomolecules were successfully immobilized and characterized optically and electrochemically. Optical immunoassay with indirect competitive ELISA were developed for the determination of BaP compound which showed the best performance compared to other assay. In optical measurement, normal assay exhibits higher absorbance, implying high bio complex formation. Modification of electrode using 11-MUA was successful due to highest blocking effect for electron transfer process. Determination of BaP in real water samples shows that there no BaP was detected in all samples collected, due to no presence of BaP in all samples.

 

Keywords:  surface modification, screen-printed gold electrode, benzo[a]pyrene, water sample

 

Abstrak

Kertas kerja ini menerangkan pengubahsuaian permukaan elektrod emas skrin tercetak menggunakan lapisan mono tepasang menggunakan tiol, dengan kombinasi format ujian enzim-pengerap imuno (ELISA) untuk penentuan benzo[a]pirina (BaP) dalam sampel air. Semua molekul bio telah dipegunkan di atas permukaan elektrod bekerja untuk meningkatkan kepekaan biosensor yang dibangunkan dengan menggunakan pelbagai jenis ELISA. Pencirian elektrokimia menggunakan voltametri kitaran (CV) dan spektroskopi impedans elektrokimia (EIS) telah dilakukan untuk mengkaji tingkah laku kekonduksian biosensor yang dibangunkan. Berdasarkan keputusan yang diperolehi, molekul bio telah berjaya dipegunkan dan dicirikan secara optik dan elektrokimia. Ujian imuno optik dengan ELISA bersaing secara tak langsung telah dibangunkan untuk penentuan sebatian BaP menunjukkan prestasi terbaik berbanding ujian lain. Dalam pengukuran optik, ujian normal mempamerkan keserapan lebih tinggi, menandakan pembentukan kompleks bio yang tinggi. Pengubahsuaian elektrod menggunakan 11-MUA berjaya disebabkan oleh kesan halangan tertinggi bagi proses pemindahan elektron. Penentuan BaP dalam sampel air sebenar menunjukkan tiada BaP dikesan dalam semua sampel yang diambil, kerana ketiadaan BaP dalam semua sampel.  

 

Kata kunci:  pengubahsuaian permukaan, elektrod emas skrin tercetak, benzo[a]pirina, sampel air

 

References

1.       World Health Organisation Regional Office for Europe (2000). Polynuclear aromatic hydrocarbons (PAH). Air Quality Guidelines for Europe. Second Edition, Copenhagen: pp. 1 – 24.

2.       Van Emon, J. M. (2007). Immunoassay and other bioanalytical techniques. Florida, USA: CRC Press.

3.       Moina, C. and Ybarra, G. (2012). Fundamentals and applications of immunosensors. In N. H. L. Chiu (Ed.), Advances in immunoassay technology. Rijeka, Croatia: pp. 65 – 80.

4.       Lewis, J. G., Clifford, J. K. and Elder, P. A. (1990). Monoclonal antibodies to pregnanediol-3-glucuronide: Application to a direct enzyme-linked immunosorbent assay of urine. Steroids, 55: 314 –318.

5.       Chung, M. K., Riby, J., Li, H., Iavarone, A. T., Williams, E. R., Zheng, Y. and Rappaport S. M. (2010). A sandwich enzyme-linked immunosorbent assay for adducts of polycyclic aromatic hydrocarbons with human serum albumin. Analytical Biochemistry, 400: 123 – 129.

6.       Chung, M. K., Regazzoni, L., McClean, M., Herrick, R. and Rappaport, S. M. (2013). A sandwich ELISA for measuring benzo[a]pyrene-albumin adducts in human plasma. Analytical Biochemistry, 435: 140 – 149.

7.       Gui, W. J., Liu, Y. H., Wang, C. M., Liang X. and Zhu, G. N. (2009). Development of a direct competitive enzyme-linked immunosorbent assay for parathion residue in food samples. Analytical Biochemistry, 393: 88 – 94.

8.       Song, J., Yang, H., Wang, Y., Si, W. and Deng, A. (2012). Direct detection of 3-amino-5-methylmorpholino-2-oxazolidinone (AMOZ) in food samples without derivatisation step by a sensitive and specific monoclonal antibody based ELISA. Food Chemistry, 135: 1330 – 1336.

9.       Perrotta, P. R., Arevalo, F. J., Vettorazzi, N. R. and Zon, M. A. (2012). Development of a very sensitive electrochemical magneto immunosensor for the direct determination of ochratoxin A in red wine. Sensors and Actuators B: Chemical, 162: 327 – 333.

10.    Van Emon, J. M., Chuang, J. C., Lordo, R. A., Schrock, M. E., Nichkova, M., Gee, S. J. and Hammock, B. D. (2008). An enzyme-linked immunosorbent assay for the determination of dioxins in contaminated sediment and soil samples. Chemosphere, 72: 95 – 103.

11.    Barcelo, D., Oubina, A., Salau, J. S. and Perez, S. (1998). Determination of PAHs in river water samples by ELISA. Analytica Chimica Acta, 376: 49 – 53.

12.    Matschulat, D., Deng, A., Niessner, R. and Knopp, D. (2005). Development of a highly sensitive monoclonal antibody based ELISA for detection of benzo[a]pyrene in potable water. Analyst, 130: 1078 – 1086.

13.    Li, K., Woodward, L. A., Karu, A. E. and Li, Q. X. (2000). Immunochemical detection of polycyclic aromatic hydrocarbons and 1-hydroxypyrene in water and sediment samples. Analytica Chimica Acta, 419: 1 – 8.

14.    Fahnrich, K. A., Pravda, M. and Guilbault, G. G. (2003). Disposable amperometric immunosensor for the detection of polycyclic aromatic hydrocarbons (PAHs) using screen-printed electrodes. Biosensors and Bioelectronics, 18: 73 – 82.

15.    Ahmad, A., Lee, P. K. and Jusoh, N. S. (2016). A comparative study of thiols self-assembled monolayers on gold electrode. Middle-East Journal of Scientific Research. 24(6):2152 – 2158.

16.    Ahmad, A. and Moore. E. (2012). Electrochemical immunosensor modified with self-assembled monolayer of 11-mercaptoundecanoic acid on gold electrodes for detection of benzo[a]pyrene in water. Analyst, 137: 5839 – 5844.

17.    Mishra, S. K., Kumar, D. and Biradar, A. M. (2012). Electrochemical impedance spectroscopy characterisation of mercaptopropionic acid capped ZnS nanocrystal based bioelectrode for the detection of the cardiac biomarker-myoglobin. Bioelectrochemistry, 88: 118 – 126.