title: Potentiometric determination of α-L-fucosidase enzyme by using 2-chloro-4-nitrophenol-rhodamine B ion pair chemical recognition in PVC membrane sensor.
abstract: The activity of the α-L-fucosidase (AFU) enzyme represents an excellent test for diagnosis of hepatocellular carcinoma (HCC) and fucosidosis recognized in inborn disorder of metabolism and increases the sensitivity of detection to 95.5% in patients with HCC. Therefore, the determination of the activity of AFU enzyme is very important and can be used as a screening tool for the early diagnosis of tumors for HCC patients. A simple, accurate, and sensitive potentiometric method was developed for measuring the activity of AFU. The method was based upon measuring the concentration of 2-chloro-4-nitrophenol (2-chloro-4-NP) using a 2-chloro-4-NP-rhodamine B ion pair in a PVC membrane sensor. The electrode shows a linear, reproducible, and stable potentiometric response with an anionic Nernstian slope of -51.13 ± 0.6 mV/decade over a wide range of concentrations 10(-5)-10(-2) M and a detection limit of 1.0 × 10(-6) M of 2-chloro-4-NP. The membrane exhibits a fast response time of 30 s, over a pH range of 4.0-6.5. The selectivity coefficients indicate excellent selectivity for 2-chloro-4-NP over a number of interfering species, e.g., chloride, nitrate, sulfate, chromate urea, albumin, glucose, uric acid, and total protein. The prepared sensor has been used successfully for the determination of 2-chloro-4-NP produced from the hydrolysis of 2-chloro-4-NP-α-L-fucopyranoside substrate. It was also applied for the determination α-L-fucosidase enzyme of 33 serum samples of healthy subjects and patients. The average recoveries ± RSD for the healthy subjects, cirrhosis of chronic hepatitis C and B, and HCC serum samples were 102.6 ± 1.01%, 101.5 ± 0.95%, and 100.1 ± 1.1%, respectively. The results obtained are in good agreement with those obtained by standard methods.
authors.affiliation: Genetic Engineering and Biotechnology institute, Menoufia University, Sadat City, Egypt. hameed.mahmoud61@hotmail.com
authors.forename: Abdelhameed M
authors.initials: AM
authors.lastname: Othman
authors.forename: M E
authors.initials: ME
authors.lastname: El-Houseini
authors.forename: Mohamed S
authors.initials: MS
authors.lastname: El-Sofy
authors.forename: Hassan Y
authors.initials: HY
authors.lastname: Aboul-Enein
mesh_terms: D000328:Adult; D000368:Aged; D015374:Biosensing Techniques / Q000191:economics / Q000379:methods*; D006528:Carcinoma, Hepatocellular / Q000201:enzymology*; D005260:Female; D019698:Hepatitis C, Chronic / Q000201:enzymology; D006801:Humans; D057230:Limit of Detection; D008297:Male; D008567:Membranes, Artificial; D008875:Middle Aged; D009596:Nitrophenols / Q000737:chemistry* / Q000378:metabolism; D011143:Polyvinyl Chloride / Q000737:chemistry* / Q000378:metabolism; D011199:Potentiometry / Q000191:economics / Q000379:methods; D012235:Rhodamines / Q000737:chemistry*; D055815:Young Adult; D005644:alpha-L-Fucosidase / Q000097:blood* / Q000378:metabolism
chemical_list: D008567:Membranes, Artificial; D009596:Nitrophenols; D012235:Rhodamines; D011143:Polyvinyl Chloride; D005644:alpha-L-Fucosidase; C029773:rhodamine B; C548837:2-chloro-4-nitrophenol