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Anal Bioanal Chem. 2013 Jan;405(1):297-305.
Shimomura T, Sumiya T, Ono M, Ito T, Hanaoka TA.
Funai Electric Advanced Applied Technology Research Institute Inc. (FEAT), TCI 37A, 2-1-6, Sengen, Tsukuba-shi, Ibaraki 305-0047, Japan. [email protected].
Abstract
A disposable amperometric biosensor for ketone 3-{Beta}-hydroxybutyrate (3HB) has been developed successfully. The sensor is based on a screen-printed carbon electrode containing Meldola’s Blue (MB) and sensing components containing nicotinamide adenine dinucleotide (NAD(+)) and 3-{Beta}-hydroxybutyrate dehydrogenase (3HBDH) immobilized in mesoporous silica (FSM8.0) using an aqueous photo-cross-linkable polymer matrix of polyvinyl alcohol (O-391), and it requires only a small sample volume of 10 uL for the measurement. The behavior of a resulting biosensor, i.e., 3HBDH-FSM8.0/NAD(+)/MB-SPCE, was examined in terms of NAD(+) concentration for construction, pH, applied potential, operational range, selectivity, and storage stability. The sensor showed an optimum response at a pH of 7.6 and at an applied potential of -50 mV. The determination range and the response time for 3HB were from 30 uM to 8 mM and approximately 30 s, respectively. In addition, the sensor was quite stable and maintained >90% of its initial response after being stored for over 6 months. This result implies that our method provides a novel biosensor for ketone 3-{Beta}-hydroxybutyrate which is easy-to-use, cost-effective, and has good reproducibility, which are vital for commercial purposes.
Additional Information
This research is based on the following patents and previous articles:
[1] T. Shimomura, et al., ‘Enzyme Electrode And Enzyme Sensor’, WO 2008/153157. [2] T. Shimomura, et al., ‘Enzyme Electrode And Enzyme Sensor’, JP 5219040. [3] T. Shimomura, et al., ‘Electrochemical biosensor for the detection of formaldehyde based on enzyme immobilization in mesoporous silica materials’, Sens. Actuators B: Chem. 135 (2008) 268−275. [4] T. Shimomura, et al., ‘Amperometric determination of choline with enzyme immobilized in a hybrid mesoporous membrane’, Talanta 78 (2009) 217−220. [5] T. Shimomura, et al., ‘Amperometric biosensor based on enzymes immobilized in hybrid mesoporous membranes for the determination of acetylcholine’, Enzyme Microb. Technol. 45 (2009) 443−448. [6] T. Shimomura, et al., ‘Amperometric detection of phenolic compounds with enzyme immobilized in mesoporous silica prepared by electrophoretic deposition’, Sen. Actuators B: Chem. 153 (2011) 361−368. [7] T. Shimomura, et al., ‘Amperometric l-lactate biosensor based on screen-printed carbon electrode containing cobalt phthalocyanine, coated with lactate oxidase-mesoporous silica conjugate layer’, Anal. Chim. Acta. 714 (2012) 114−120. [8] T. Shimomura, et al., ‘An electrochemical biosensor for the determination of lactic acid in expiration’, Procedia Chemistry 6 (2012) 46−51.
So far, we have developed high-performance enzymatic sensors that are based on enzyme encapsulation by mesoporous silica material. For example, we have successfully developed various high performance aqueous and gaseous sensors (e.g. residual pesticides, phenolic compounds, alcohol and ammonia) which can detect ultra-low concentration of target substances quickly. Based on this highly advanced sensing technique, one can obtain a novel biosensor which is easy-to-use, cost-effective, and with good reproducibility, which are vital for the commercialization of the biosensor.
Figure Legend
Image. Schematic diagram of detection mechanism of our enzymatic sensor.
