Biosensors and Bioelectronics 24 (2009) 10831089

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Biosensors and Bioelectronics

journal homepage: www.elsevier.com/locate/bios

Review

The evolution of commercialized glucose sensors in China

Jamie Hu
Shanghai Bioscan Inc., 203, Gumei Business Center, 1905 Long Ming Road, Shanghai 201101, China

a r t i c l e

i n f o

a b s t r a c t
The glucose monitor with a screen-printed carbon sensor has been in commercial production since 1994. In last 15 years, around 10 companies have been involved in manufacturing and marketing the meters and glucose test strips and are being strong competitors of the companies which import these products. Comparison of early stage glucose meters and glucose test strips with latest fabrications showed a large increase in production volume and improved functional features. It also showed technological development of glucose monitors including circuit improvement, as more integrated computer processor units (CPU) are now being used. The technology of mass-production of disposable screen-printed test strips has been widely used in local industries mainly for the production of blood glucose test strips. The opportunities and challenges in local diabetes market are discussed in this paper. 2008 Elsevier B.V. All rights reserved.

Article history: Received 31 March 2008 Received in revised form 20 August 2008 Accepted 28 August 2008 Available online 10 September 2008 Keywords: Enzyme sensors Screen-printed carbon-based electrodes Mediators Glucose monitors

Contents 1. 2. 3. 4. 5. 6. 7. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mediator-modied SPCEs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Glucose enzyme-modied SPCEs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Evolution of glucose testing meter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The local market share for blood glucose monitoring systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The challenges for local companies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1083 1084 1084 1087 1087 1088 1088 1089 1089

1. Introduction According to the mid of 2006 report from Ministry of Health, nearly 40 million people suffer from diabetes in China (Chin CDC). Since diabetic patients need to control their blood glucose levels carefully, the importance of self-monitoring of blood glucose has been widely recognized as an effective method of measuring blood sugar not only in clinics but also at home and in the working place. The rst research on ampreometric determination of blood glucose using a redox couple-mediated, glucose oxidase (GOx)-catalyzed reaction was demonstrated by Williams et al. (1970). But this study did not lead to the rapid application of amperometry in self-monitoring blood glucose in the home.

Tel.: +86 21 51560919; fax: +86 21 64800290. E-mail address: jh@bioscan.net.cn. 0956-5663/$ see front matter 2008 Elsevier B.V. All rights reserved. doi:10.1016/j.bios.2008.08.051

The rst electrochemical blood glucose monitor for selfmonitoring diabetic patients was pen-sized and was launched in 1987 as ExacTech by Medisense Inc. Many studies on the development of blood glucose sensors have been carried out since the 1980s (Cass et al., 1984; Newman et al., 1992; Wang et al., 1995; Cui et al., 2000; Gao et al., 2005; Newman and Turner, 2005; Heller and Feldman, 2008). With regard to the huge local requirement of millions of diabetes suffers, the production of glucose sensors and test meters is still growing fast and many of the methods being developed have been based on electrochemical techniques such as the use of carbon electrodes, and are particularly attractive for this have been based on the use of electrochemical techniques such as carbon electrodes are particularly attractive for this application due to their high sensitivity, selectivity, small size and low cost (Hu, 1989). Screen-printing of carbon ink for the fabrication of electrochemical sensors has realized commercial success in glucose sensing eld. In 1993, Hu was the rst to use screen-printing tech-

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nology for the mass production of extremely inexpensive, highly reproducible and reliable single-use glucose sensors in China. From this point on printing technology in the production of disposable low-cost enzyme sensors for the amperometric determination of blood glucose has been widespread growth in this country. Several hundreds of basic research papers have been published in China on electrochemical glucose sensors since 1990s (Dong et al., 1991; Hu and Turner, 1991; Jin et al., 1994; Zhang and Rechnize, 1994). Because of this large number of publications, a full review of the literature, even of the most recent advances, is difcult to achieve. Nevertheless this paper intends to acquaint the reader with the fundamentals of the electrochemistry of glucose and provide a perspective of the evolution of the electrochemical glucose assay and monitors in China where 40 million people suffer from diabetes. 2. Mediator-modied SPCEs Rapid and low interference analysis of blood sugar is of great interest in the diagnosis of diabetes. A large number of the disposable enzyme sensors used in the analysis of glucose are based on electrochemical determination of enzyme-generated hydrogen peroxide as shown in the following reaction: Glucose + O2 + Glucoseoxidase Gluconicacid + H2 O2 In this reaction, oxidation and reduction of H2 O2 generally requires a high potential at the bare metal electrodes, which implies a large interference. For this reason, most of the portable glucose testing systems with glucose sensors use mediators that enable the reduction at low potential, thereby decreasing the electrochemical interference. Due to the simple technological processing and low cost, graphite products are preferred to be the base material for screen-printed carbon electrodes for glucose determination (Hu and Ge, 1999). Different carbon inks obtained from Acheson, DuPont, Gwent and Ercon have been compared for the production of base electrodes in glucose test strips. Fig. 1 shows the cyclic voltammograms of the ferri/ferrocyanide redox couple at the commercial carbon strip electrodes screen-printed with the four carbon inks mentioned above. Of the commercial sensors, On-Call-Plus using DuPont carbon ink yielded a best reversibility as Fig. 1(a) but the other commercial electrodes were also found to be effective. Fig. 1(c) and (d) shows the effect of commercial electrodes from Yicheng and Jinque which showed high peaks at the 400 and 300 mV, respectively, and were designed using silver as the conducting tracks. The rst enzymatic carbon screen-printed glucose sensor using Acheson inks was developed by Hu in early of 1994 (Hu and Ge, 1999). In this sensor, glucose oxidase was adsorbed onto the surface of the carbon electrode and benzoquinone was used as a mediator. The reactions at the electrode are as follows: Glucose + Quinone + Glucoseoxidase Gluconicacid + Hydroquinone Hydroquinone Quinone + 2H+ + 2e The rst blood glucose testing system was launched in China in 1994 and was based on the results mentioned above. Later, Hu and Zhang used ferricyanide as the mediator because ferricyanide is more soluble and sensitive than benzoquinone in the water phase. This reaction is as follows: Glucose + Ferricyanide + Glucoseoxidase Gluconicacid + Ferrocyanide + H 2 O2

H2 O2 2H+ + 2e Initial cyclic voltammetric investigations of ferricyanide adsorbed onto a carbon-based electrode showed promising results for the electro-catalytic oxidation of glucose and glucose oxidase. (Zhang et al., 1999; Hu and Ge, 1999). In response to the growing market demand, researchers undertook studies to nd new mediators with fast reaction times and lower interference. Yicheng recently tested SPCEs with hexaamineruthenium(III) chloride mediator as the electron shuttle. The company found that the use of this mediator could eliminate interference from other oxidizable species in the blood, providing improved analytical results in the determination of blood glucose and leading to a precise test strip which required only a 0.5 l sample of blood and produced the results in 5 s (Yicheng Bioelectronics Ltd., www.yichengbioelectronics.com.cn.2007/) (Chinese). Another mediator osmium (II)-poly-pyridine was also used in the construction of screen-printed carbon-based electrodes, and was used mixed with the enzyme glucose oxidase to form a solution which was dispensed onto the SPCEs creating a thin layer after drying at 55 C for 5 min. A company in Guilin has shown that the test strip containing the osmium mediator could be successfully applied to the analysis of blood glucose in clinical practice. (Guilin Zhonghui Technology Co., Ltd., www.glzhkj.cn2007/) (Chinese) 3. Glucose enzyme-modied SPCEs Glucose oxidase is an oxygen-dependent enzyme which has long been used in the production of glucose sensors. However, there are some problems associated with the use of this enzyme for glucose monitoring. As the enzyme uses oxygen as an electron acceptor, there is a competition between the articial mediator and oxygen mediations. Thus, if the blood oxygen level is high, measurement of blood glucose may appear lower than the true value. For this reason, some other enzymes such as pyrroloquinoline-quinonedependent glucose dehydrogenase (PQQ-GDH) have been used in the construction of disposable glucose test strips. In this system the blood oxygen level does not affect reaction, as the enzyme does not use oxygen as an electron acceptor. The PQQ-GDH enzyme is also superior in its reaction speed to glucose oxidase. According to the recent research and substantiated in some cases, the pyrroloquinoline-quinone-dependent glucose dehydrogenase (PQQ-GDH) is not sufciently substrate specic as it also reacts with maltose, galactose and maltotriose (Tsujmura et al., 2006; Tamadaka and Soda, 2007). This enzyme prole has led to a report of errors in the results when the PQQ-GDH enzyme glucose test strip was used to monitor blood glucose in diabetic patients. Recently, the enzyme called avin adenine dinucleotidedependent glucose dehydrogenase (FAD-GDH) (Amano Coxmpany Report, 2007, www.amano.com.jp/) was shown to have thermostability and high substrate specicity. This FAD-dependent glucose dehydrogenase would therefore be superior to other glucose oxidoreductase tested to date in the construction of an O2 -insensitive amperometric glucose sensor. The two enzymes mentioned above have been of great interest to local companies and have been used for pilot scale production of glucose sensors (Hu, in press). Fig. 2 shows the measurement of blood glucose at a concentration of 400 mg/dl using FADGDH/ferricyanide-modied glucose test strips and an operating potential of 300 mV. The current peak was reached in about 0.7 s. In order to improve the properties of the disposable glucose sensors use glucose oxidase and glucose dehyrogenase, new materials and improved screen-printing techniques have been developed in recent years. Hu demonstrated the procedure of fabricating a disposable amperometric glucose sensor using water-based enzyme

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Fig. 1. Cyclic voltammograms for different electrodes in 50 mM ferri/ferrocyanide and 100 mM KCl support electrolyte. Scan rate at 100 mV/s. (a) DuPont carbon ink, (b) Acheson carbon ink, (c) Gwent carbon ink and (d) Ercon carbon ink.

Fig. 2. Current response of the glucose sensor based on time, the FADGDH/ferricyanide was screen-printed onto the working area of the strip. A potential of +300 mV vs. the reference/counter electrodes was applied for 3 s. The sample contained whole blood glucose with a glucose concentration of 400 mg/dl which had been calibrated by YSI-2300 plus, the hematocrit was adjusted to 40%.

ink containing binder, stabilizer, mediator and surfactant. This type of the enzyme ink presents a great advantage: it avoids problems of enzyme denaturation as it is not necessary to employ organic solvents, and the curing temperature is therefore lower. It enables the production of glucose test strip to be increased to half-million pieces per day, which leads to a considerable reduction in operation cost. This processing technology for large-scale production of enzyme sensor has been successfully commercialized in local industries (Hu CN Patent, 2007). Since 1993, glucose test strips made in this country have been updated to produce several generations of this product; the following gures show the recent history of test strip development (Hu, 2004). Fig. 3 shows the test strips made in 1990s, in this case, a carbonworking electrode and a silver/silver chloride reference electrode system was designed, the operation voltage was set at 400 mV. Enzyme and mediator were deposited on the surface of electrode with a dispenser. A large volume of blood (20 l sample) was required and the test time was around 2025 s and the sample had to be applied to the top centre of test strip surface.

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Fig. 3. Products made in early of 1990s, blood glucose test strips were designed with one carbon working electrode and one Ag/AgCl reference electrode and the reaction area was covered by a mesh to ensure uniform dispersion of the blood sample.

Fig. 5. Test strips made in 2005, the size of the capillary chamber had been further reduced (1 l). The strip chemistry consisting of an enzyme, a new redox mediator and other components were screen-printed over the surface of electrodes and often on an entire side of the capillary chamber.

Fig. 4 shows the test strip made in late of 1999 which also uses two electrodes; however, in this case both electrodes consisted of a carbon-based material. The operation voltage was set at 300 mV and the blood sample was automatically lled into reaction area at edge side of strip by a capillary mechanism. The reaction area was dened by a spacer and a hydrophilic membrane cover, thus the sample volume entering the reaction site was reduced to 3 l. Test time was also greatly reduced to 10 s.

Fig. 5 shows an example made in mid-2000; although it was a two-electrode design the quality of screen-printing was much improved, the sample volume was further reduced to 0.52 l, and the test time was shortened to 5 s. New mediators were used in attempt to eliminate the interference from the blood. Fig. 6 shows an example of the latest test strips which contain more electrodes (three electrodes) that are adapted and printed on the support, showing that local manufacturers are aiming at improving test accuracy and minimizing interference from blood sample. In this design, one electrode acts as a trigger which gives an exact timing signal to the meter. The sample has been repositioned at the centre of the top edge as shown in the gure, facilitating use by patient and avoiding operational errors. Several ingenious combinations of elements have been considered: (1) a plastic substrate with thermostability was selected to give improved curing of carbon and silver pastes; (2) working and reference/counter electrodes

Fig. 4. Carbon/carbon test strip design, a small volume (3 l) capillary chamber was formed over the plastic substrate and the electrodes were attached often by means of a spacer such as a pressure sensitive adhesive and a hydrophilic cover layer.

Fig. 6. Test strips designed with three electrodes; R/C, W, and F indicate the reference/counter, working and ll electrodes respectively.

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Fig. 7. Commercial meters designed and manufactured in the early 1990s; the test time was over 25 s and more than 20 l of the blood sample was required. From the left to right, devices produced by Yicheng, Jinque and Jiaji respectively.

Fig. 8. Latest designs of glucose test meters with test times around 5 s and a sample size requirement of only 1 l blood. Devices manufactured by Sannuo, Acon and Yicheng are shown from left to right respectively.

were positioned close proximity to decrease the resistance of the system solution in order to include on-strip hematocrit compensation; (3) the screen-printable water-based enzyme paste was used to lower the production costs; (4) ll detection triggers which are vestigial electrodes were employed to enable the meter to detect when the strip is sufciently lled with blood and to initiate the assay as soon as the chamber is lled; and (5) very thin spacer design allowed the use of near- or sub-microliter blood samples. 4. Evolution of glucose testing meter A portable glucose testing meter should be functional, compacted, user friendly and produced at low cost. Fortunately with the rapid development of the microelectronic industry in the local area, these requirements are easily fullled in the scale-up production of glucose meters. Figs. 7 and 8 show the various stages in the manufacture of glucose meters by local companies. Handheld glucose meters, as shown in Fig. 7, have been under development in China since the early 1990s. At this time there was less choice of computer processor unit (CPU) chips, the circuitry was complex and less integrated and the nished machine was rather bulky. Fig. 7 shows some of the machines manufactured in 1994 and in these devices all the components including the CPU, LCD Driver, A/D converter, amplier, charge pump were separated. These meters were capable of only simple functions and could store a maximum of 10 readings, and were therefore unable to provide a screen display of the time, date and average blood test results in 7-, 14- and 28-days periods (Hu, 2007a). Fig. 8 shows latest design, which has a more integrated CPU with the liquid display driver (LCD), amplier, analog/digital (A/D)
Table 1 Features of local brand glucose monitoring systems Meters Coding Sample volume (l) Length of test time (s) Range (mmol/l) Precision (%) Memory Data Average Data download Control solution Enzyme deposit Enzyme Mediator Battery Strip Package Strip expires (months) Yicheng Build-in 0.5 5 2.227.8 CV < 7.5 10 stored No No No Screen-printed GOx Ferricyanide Lithium Single 12 Sino Build-in 3 25 2.227.8 CV < 7.5 50 stored Yes No Yes Dispense GOx Ferricyanide AAA Single 12

converter and charge pump incorporated into a single chip. This design resulted in a compact easily manageable device and led to greatly reduced production cost. A smart CPU named HOLTEK 56*64 was selected, it is 8-bit, high performance, RISC architecture microcontroller device specically designed for A/D product applications that interface directly to analog signals and which require an LCD Interface. The HT56*64 mask version device is 64pin and functionally compatible. The advantages of low power consumption, I/O exibility, timer functions, oscillator options, 12channel A/D converter, pulse width modulation function, power down and wake-up functions, in addition to a exible and congurable LCD interface enhance the versatility of the device to control a wide range of applications requiring analog signal processing and LCD interfacing, especially for blood glucose metering (Hu, 2007b; www.aconlab.com.cn2007/) 5. The local market share for blood glucose monitoring systems Blood glucose monitors have the appearance of uncomplicated devices about the size of a cell phones, uncomplicated devices. However, they are extremely intricate machines that use the latest technological advances (both in meters and strips) to provide an accurate and easy manage test of blood glucose levels. Competition between manufacturers of these products was the driving force that led to the changes in design. Any new innovation or useful feature of the meter or strip that was developed by one company was analyzed by other companies to establish whether it was worth emulating. Companies strive to gain a competitive advantage by continuing updating their product line in order to attract purchasers. As a result

Jinque Build-in 3 20 2.227.8 CV < 7.5 30 stored No No No Dispense GOx Ferricyanide AAA Bottle 12

Jiaji Build-in 3 20 2.227.8 CV < 7.5 30 stored No No No Dispense GOx Ferricyanide Lithium Bottle 12

Acon Code chip 1 10 1.133.3 CV < 5.5 300 stored Yes Yes Yes Dispense GOx Ferricyanide Lithium Bottle 18

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Fig. 9. Local market share of glucose monitoring systems; total 19.3 million (USD) in 2006 (The Market survey of glucose monitoring system in China, 2007. www.allmyinfo.com/eng/report). (For interpretation of the references to color in the artwork, the reader is referred to the web version of the article.)

Fig. 10. shows the local sales market in 20052006 and estimates the sales from 20072010 (The Market survey of glucose monitoring system in China, 2007. www.allmyinfo.com/eng/report). *Values in million RMB; 1U$ = 7 RMB.

of this rapidly changing market, a meter that is state-of-the-art in 2007 may be outdated by 2008. There are about 10 companies in local area producing glucose meters and strips. Table 1 lists ve of ten local brand blood glucose testing systems and product functions are also listed. These companies shared only 510% of the local market. The total sales of glucose monitoring system in 2007 amounted to around $400 million. This gure still represents low sales of these devices considering that there are 40-million diabetes in mainland China as compared with the US and European countries. Fig. 9 shows the market shares of local brands in 2006, with Yicheng Co., Ltd. having 35%, Sino and Acon both around 20%, Jinque only 3% and others accounting for 22% of the total. Although the sales volume of local brands is small at present, they have the advantages of low prices and good services and therefore are seen as serious competition to foreign importers in the market. Local manufacturers increased their sales by 2030% over the last 3 years and hopefully local products will account 1015% of market sales by the end of 2010 and will take an even bigger share of market by 2015. Fig. 10 shows the survey of the market for locally produced glucose monitoring systems. 6. The challenges for local companies From the technological point of view, the local companies are still facing some hurdles which need to be overcome. The screenprinting processing technology for mass production of test strips is still in need of improvement and lacks strict quality control checks throughout the production line, as automatic manufacturing and conveyers are not widely used and most production processes are still carried out by hand. This results in low accuracy and poor reproducibility of test strips.
Table 2 Domestic products compared with some imports on technological issues Brand/features Sample volume (l) Test time (s) Precision (%) Hematocrit (%) Correlation efcient ( ) Enzymes used Mediators Enzyme loading Strip need coding CE/FDA Approval Free style 0.3 5 CV 3 2070 0.98 GDH Os complex Screen-printing Yes Yes Bayer 0.6 5 CV 5.3 1565 0.98 GDH Ferricyanide Screen-printing No Yes

The management is poor, some international recognised standards such as ISO13485; ISO 151972003 and also good manufacture practise (GMP) are not adhered by these companies. To day only three of 10 Chinese companies have achieved CE Certication and none has been given US Food and Drug Administration (FDA) approval. This explains why the Chinese glucose testing products are rarely found in the international market. Table 2 compares the technical characters of local brands with those of imported products found in local market. It also shows that international brand blood glucose testing systems have better precision than local products. 7. Conclusions In recent years, there has been rapid development in the technology required to produce screen-printed carbon-based electrodes for application to the analysis of blood sugar as evidenced by the large volume of research that has been reported. The types of sensors or strips described in this work will contribute to satisfying the growing demand for rapid and accurate in situ analyses and the development of compact and portable devices. This study has endeavoured to review the current situation concerning the development and production of glucose monitoring systems in China. Although local manufacturers are facing some problems with regard to both technological issues and management, signicant improvements in their products are expected to be achieved mainly by incorporating new processing technology, new printed materials and improving printing technology to lead to the enhancement of reproducibility and sensitivity of screen-printed carbon-based strips. With this aim, new research to modify the working and counter electrodes is ongoing, with the focus on new

Lifescan 1 5 CV 4.4 2070 0.98 GOx Ferricyanide Screen-printing Yes Yes

Yicheng 0.5 5 CV < 7.5 3555 0.95 GOx Ferricyanide Screen-printing Yes No

Acon 1 10 CV < 5.5 3555 0.97 GOx Ferricyanide Dispensing Yes CE only

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enzymes, new mediators, conducting polymers and nanostructure materials. Acknowledgements The work was partially supported by local manufacturer Yicheng Bioelectronics Ltd. in Beijing. The author has been engaged in useful discussion with Acon Lab in Hangzhou. Thanks are also due to Dr. Shaohua Zuo for data collection. References
Cass, A.E.G., Davis, G., Francis, G.D., Hill, H.A.O., Aston, W.J., Higgins, I.J., Plotkin, E.V., Scott, L.D.L., Turner, A.P.F., 1984. Anal. Chem. 56, 667671. Center for Disease Control (Ministry of Health, China www.eds.org.cn/) 2005, Guideline for Management of Chinese Diabetes. Cui, G., Kim, S.J., Choi, S.H., Nam, H., Cha, G.S., Paeng, K.J., 2000. Anal. Chem. 72, 1925. Dong, S., Wang, B., Liu, B., 1991. Biosens. Bioelectron. 7, 215220.

Enzyme Wave 2007, 10. www.amano-enzyme.co.jp/. Gao, Z.Q., Xie, F., Shariff, M., Arshad, M., Yin, J.Y., 2005. Sens. Actuators B111, 339345. Heller, A., Feldman, B., 2008. Chem. Rev. 108, 24822505. Hu, J., 1989. Chin. J. Biotechnol. 6, 328331. Hu, J., Ge, B.X., 1999. Adv. Biosens. 4, 315325. Hu, J., Turner, A.P.F., 1991. Anal. Lett. 24, 1524. Hu, J., 2004. Chin. Chem. Sens. 24, 18. Hu, J., 2007a. Chin. Sens. World 6, 1013. Hu, J., 2007b. Chin. Chem. Sens. 27, 2126. Hu, J., 2007. CN Patent 200710037552X (Chinese). Hu, J., in press. Chin. Chem. Sens. (Chinese). Jin, L., Mao, Y., Sun, X., Fang, Y., 1994. Anal. Lab. 13, 1317. Newman, J.D., Turner, A.P.F., 2005. Biosens. Bioelectron. 20, 24352453. Newman, J.D., Turner, A.P.F., Marrazza, G., 1992. Anal. Chim. Acta 262, 1316. Tamadaka, H., Soda, K., 2007. J. Diab. Sci. Technol. 1, 2836. Tsujmura, S., Kojima, S., Kano, K., Ikeda, T., Sato, M., Sanada, H., Omura, H., 2006. Bios. Biotech. Biochem. 70, 654659. Wang, J., Chen, Q.A., Pedrero, M., Pingarro, J.M., 1995. Anal. Chim. Acta 300, 111. Williams, D.L., Doig A.R.Jr., Korosi, A., 1970. Anal. Chem. 42, 118123. Zhang, X., Rechnize, G.G., 1994. Electroanalysis 6, 361370. Zhang, Z.R., Zhang, G.X., Zhu, J.Z., Hu, J., 1999. Adv. Biosens. 4, 273287.