Microchimica Acta

, Volume 174, Issue 3–4, pp 303–309 | Cite as

Biosensor based on a glassy carbon electrode modified with tyrosinase immmobilized on multiwalled carbon nanotubes

  • Jing Ren
  • Tian-Fang Kang
  • Rui Xue
  • Chao-Nan Ge
  • Shui-Yuan Cheng
Original Paper

Abstract

We describe a biosensor for phenolic compounds that is based on a glassy carbon electrode modified with tyrosinase immobilized on multiwalled carbon nanotubes (MWNTs). The MWNTs possess excellent inherent electrical conductivity which enhances the electron transfer rate and results in good electrochemical catalytic activity towards the reduction of benzoquinone produced by enzymatic reaction. The biosensor was characterized by cyclic voltammetry, and the experimental conditions were optimized. The cathodíc current is linearly related to the concentration of the phenols between 0.4 μM and 10 μM, and the detection limit is 0.2 μM. The method was applied to the determination of phenol in water samples.

Figure

A tyrosinase and carbon nanotubes (MWNTs) modified glassy carbon electrode was fabricated and used for the sensitive detection of phenol. The reduction peak of benzoquinone produced by enzymatic reaction of phenol was greatly enhanced due to the presence of MWNTs(c)

Keywords

Tyrosinase Multiwalled carbon nanotubes Electrochemical biosensors Phenols 

Introduction

Phenolic compounds often exist in the wastewaters of many industries. Many of them are very toxic, showing adverse effects on animal and plants [1]. High levels of phenols have detrimental effects on animal health. Prolonged oral or subcutaneous exposure causes damage to the lungs, liver, kidney and genitourinary tract [2]. Therefore, the determination of phenolic compounds is of great importance. Spectrophotometry and chromatography are commonly used to determine phenols [3, 4]. However, these methods are complicated in sample pretreatment and unsuitable for in situ monitoring. Biosensor based on tyrosinase (Tyr) have been proven to be a promising method for a simple, fast and sensitive detection of phenolic compounds [5, 6, 7, 8, 9, 10].

Tyrosinase can catalyse two reactions: ortho-hydroxylation of phenols to catechol and the further oxidation of catechols to ortho-quinones, both in the presence of molecular oxygen. The phenol can be detected via the reduction of the produced quinone at low potential [11, 12, 13].
$$ {\text{phenol}} + {\text{tyrosinase}}\left( {{{\text{O}}_2}} \right) \to {\text{catechol}} $$
(1)
$$ {\text{catechol}} + {\text{tyrosinase}}\left( {{{\text{O}}_2}} \right) \to {\text{o}} - {\text{quinone}} + {{\text{H}}_2}{\text{O}} $$
(2)
$$ {\text{o}} - {\text{quinone}} + {2}{{\text{H}}^{+} } + {2}{{\text{e}}^{-} } \to {\text{catechol}} $$
(3)

Recently many tyrosinase biosensors for phenols have been presented. Various materials such as magnetic nanoparticles [14], metal nanoparticles [15, 16] and polymers [12, 17] have been used to fabricate tyrosinase biosensors. Many methods have been employed for stabilizing and immobilizing the enzyme. Polymer entrapment [18, 19], electropolymerization [20, 21], sol–gels [22, 23, 24], self-assembled monolayers [25, 26], covalent linking [2, 27] and incorporation within carbon paste [11, 16, 28] have been used to construct tyrosinase biosensors. Although these materials have their own advantages in enzyme immobilization, they also have some drawbacks. Take silica sol–gel films as a example, it can retain the catalytic activities of enzymes to a large extent. But the fabrication of silica sol–gel films is time consuming and the silica sol–gel matrix is fragile, it easily shrinks, cracks and delaminates from the electrode surface [29]. Many methods which have been reported about biosensor fabrication are time consuming and consist of many steps, which result in difficulties in obtaining sensors with comparable sensitivity. Thus, more efforts are needed to develop a simple and reliable method to fabricate biosensors.

Nanomaterials possess extreme small size, a high specific surface area, a high surface-to-volume ratio and unique physicochemical characteristics. The use of nanomaterials superstructures for the creation of electrochemical devices is an extremely promising prospect. They hold important applications as catalysts. Carbon nanotubes (CNTs), as a new form of carbon, are of great interest for many applications in, for example, batteries and chemical sensors [30, 31, 32, 33]. Furthermore, high accessible surface area, low resistance and high stability suggest that CNTs are suitable for the potential catalyst supports [34, 35, 36]. After acid treatment, CNTs possess more active sites. CNTs can promote the electron transfer of the active site of biological molecules, and improve relative activity of the enzyme.

Recently, there has been growing interest in using CNTs to fabricate biosensors. Korkut et al. reported an amperometric biosensor based on horseradish peroxidase and CNTs/polypyrrole nanobiocomposite film on a gold surface for determination of phenol derivatives [37]. Yin et al. fabricated an amperometric biosensor based on tyrosinase immobilized onto multiwalled carbon nanotubes-cobalt phthalocyanine (MWNTs-CoPc) silk fibroin film to determine bisphenol A [38].

We describe sensitive biosensors for phenols here which based on tyrosinase immobilized onto MWNTs modified glassy carbon electrodes (Tyr/MWNTs/GCE). The novel characteristics of the biosensors are simple fabrication, excellent stability and high sensitivity for the determination of phonlic compounds. The characterization and performance of this tyrosinase biosensor was studied. Additionally, the biosensor has been successfully used for the detection of phenol in real samples.

Experimental

Reagents

Tyrosinase (EC 232-653-4, 3933 unit·mg−1 from mushroom) was purchased from Aldrich (http://www.sigmaaldrich.com, USA). Multiwalled carbon nanotubes (95%) were purchased from Shenzhen Nanotech Port Co. Ltd. (http://www.seasunnano.com, China). Pretreatment of MWNTs was performed by refluxing in HCl + HNO3 solution (3:1) at 110 °C for 8 h, followed by filtrating and washing with doubly distilled water till pH 7.0, dried at 110 °C for 2 h. After then, 1.0 mg MWNTs was dispersed in 10 mL DMF with the aid of ultrasonic agitation. 0.1 mg ·mL−1 treated MWNTs suspension was therefore obtained [34]. Phosphate buffer solution of pH 7.5, 0.1 M was prepared using K2HPO4 and KH2PO4 in redistilled water. All of the chemicals were of analytical reagent grade and all the solutions were prepared with redistilled deionized water.

Apparatus

All electrochemical measurements were performed with CHI842B electrochemical analyzer (Shanghai Chenhua Co., China) with a conventional three-electrode cell. A glassy carbon electrode (GCE) with diameter of 3 mm was used as working electrode, saturated calomel electrode (SCE) as reference electrode, and a platinum wire was used as auxiliary electrode.

Preparation of the electrodes and amperometric measurements

The bare GCE was polished successively with 0.5 μm Al2O3 slurry to a mirror, and ultrasonically cleaned in ethanol and water for 5 min, respectively, then washed with redistilled deionized water and dried in air before use. A 2.0 μL aliquot of the treated MWNTs solution was cast on the GCE surface, dried in air. The chemically modified electrode was denoted as MWNTs/GCE.

To dissolve 1.2 mg of tyrosinase (3,933 units·mg−1) and 3.6 mg of bovine serum albinum (BSA) in 0.2 mL phosphate buffer solution, 10 μL of this solution was mixed with 2 μL of 2.5% glutaraldehyde. 4 μL of this mixture was then deposited upon the MWNTs/GCE surface and allowed to cross-link to dryness at room temperature. This enzyme activity was calculated and it approximately equal to 80 units on each electrode surface. The electrode was denoted as Tyr/MWNTs/GCE. For comparison, Tyr/GCE was fabricated with the same method. When not in use, the biosensor was stored at 4 °C in a refrigerator.

Amperometric experiments were carried out in an electrochemical cell containing 10 mL of 0.1 M pH 7.5 phosphate buffer solution. All measurements were performed at room temperature.

Results and discussion

Electrocatalysis of MWNTs modified electrodes to quinone

The biosensor based on MWNTs with immobilized tyrosinase was first characterized using cyclic voltammetry (CV) to test its response to phenol. The immobilized tyrosinase in the presence of molecular oxygen can catalyse the oxidation of phenol to form benzoquinone. Cathodic potential scan was used to measure the reduction current of benzoquinone. The experimental results indicated that the electrochemical reduction peak of the quinone was sensitive. The reduction peak current increased with the amount of phenol added. Figure 1 shows cyclic voltammetric responses of Tyr/MWNTs/GCE in absence of phenol(a) and in presence of phenol (c). Moreover, cyclic voltammetric responses of phenol at Tyr/GCE (b) is also showed for comparison. No reduction peak for quinone was observed when the phenol is absence. After phenol solution was added into Phosphate buffer solution, benzoquinone, the enzymatic reaction product of phenol, at both Tyr/MWNTs/GCE and Tyr/GCE appeared as a reduction peak. But the reduction peak of benzoquinone at Tyr/MWNTs/GCE was much higher than that at Tyr/GCE, the peak potentials shifted positively from −0.08 V at Tyr/GCE to −0.05 V at Tyr/MWNTs/GCE, which could be attributed to that MWNTs possess excellent catalytic activity towards the electroreduction of benzoquinone. Tyr/MWNTs/GCE was, therefore, chosen as the working electrode in the following research.
Fig. 1

Cyclic voltammograms of a Tyr/MWNTs/GCE in absence of phenol in 0.1 M phosphate buffer solution (pH 7.5), b Tyr/GCE and c Tyr/MWNTs/GCE in 0.1 M Phosphate buffer solution (pH 7.5) containing 0.1 mM phenol at a scan rate of 50 mVs−1

The effect of the amounts of immobilizing tyrosinase and MWNTs on the response current

The effect of enzyme loading on the biosensor from 20 U to 120 U was investigated. As shown in Fig. 2, the current increased with the increase of the amount of tyrosinase immobilized on the electrode up to 80 U. The phenomena are similar to those previously reported [24]. When the amount of tyrosinase was more than 80 U, the current decreased with the increase of the enzyme amount. The facts can be attributed that the increase of the thickness of enzymatic film will lead to an increase of interface electron transfer resistance and make the electron transfer more difficult [39]. Therefore, 80 U tyrosinase immobilized on the modified electrode was chosen for all subsequent experiments.
Fig. 2

Effect of enzyme amount on the current response of the biosensor to 0.1 mM phenol in 0.1 M phosphate buffer solution (pH 7.5). Other conditions are defined in Fig. 1

The effect of MWNTs amount on the on reduction peak current of phenol in 0.1 M phosphate buffer solution (pH 7.5) was shown in Fig. 3. The amount of tyrosinase immobilizing on each electrode modified with MWNTs was 80 U. It can be seen that the current increased as the MWNTs amount from 0 μg to 0.2 μg, following an obvious decrease when the MWNTs amount was increased from 0.2 μg to 0.4 μg. The maximum response was obtained when the biosensor was prepared with 0.2 μg MWNTs. The reason may be that with the amount of MWNTs increasing, the enrichment active sites on electrode surface increase, leading to the current increases. However, when the amount is more than 0.2 μg, the peak current decreased, the reason may be that the thicker film of MWNTs on the electrode surface increases the diffusion distance of quinone, hinder the mass transport and charge-transfer rate at electrode. So 0.2 μg MWNTs was used to modified each electrode.
Fig. 3

Effect of multiwalled carbon nanotubes amount on the current response of the biosensor to 0.1 mM phenol at the biosensor. Other conditions are defined in Fig. 1

The effect of pH and temperature of the solution on the response current

The effect of pH value on the current response of the biosensor to 0.1 mM phenol in 0.1 M phosphate buffer solution was investigated at the pH range from 5.0 to 9.0 and the results were shown in Fig. 4. It can be seen that the reduction peak current gradually increased with an increasing pH value from 5.0 to 8.0. However, the reduction peak current decreased when the pH further increased to 9.0. At low pH range, the increase in the current response with an increasing pH could be attributed to the activity increase of tyrosinase. The optimum biosensor response was obtained at pH 8.0, which was similar to the optimum pH range of 5.0–8.0 reported for free tyrosinase [40]. This indicated that the immobilization procedure has not altered the activity of tyrosinase. Therefore, 0.1 M pH 7.5 phosphate buffer was chosen as the supporting electrolyte solution in this work.
Fig. 4

Effect of pH on the current response of the biosensor to 0.1 mM phenolin 0.1 M phosphate buffer solution at the biosensor. Other conditions are defined in Fig. 1

Since the catalytic activity of enzymes can be affected by temperature [41], the relationship between temperature and response current of the biosensor is examined in 0.1 M phosphate buffer solution (pH 7.5) containing 0.1 mM phenol. The results are displayed in Fig. 5. With an increasing temperature from 5 to 25 °C the response increased, afterwards the response decreased as the temperature further increased. The maximum response was obtained at 25 °C. Therefore, further measurements were performed at room temperature (25 °C).
Fig. 5

Effect of temperature on the current response of the biosensor to 0.1 mM phenol at the biosensor. Other conditions are defined in Fig. 1

Linear range and detection limit

Amperometry was used to estimate the detection limit of phenol. Figure 6 showed a typical amperometric current–time response of the biosensor under the optimum experimental conditions after successive addition of phenol to phosphate buffer solution under stirring every 150 s. With the increase of phenol concentration, the biosensor could achieve the steady state current within less than 20 s, which indicated a fast electron transfer process. The current linearly increases with the concentration of phenol from 4.0 × 10−7 to 1.0 × 10−5 M and the regression equation can be expressed as i(μA) = 0.2261c(μM)−0.0758, with a correlation coefficient of r = 0.9995. The detection limit was estimated to be 2.0 × 10−7 M (S/N = 3).
Fig. 6

Current–time response curve of the biosensor upon successive additions of phenol with different concentrations. Applied potential: −0.05 V

In order to know the response of the biosensor for other phenolic compounds, the experiments on other three phenolic compounds (hydroquinone, catechol and bisphenol A) were also performed by the Tyr/MWNTs/GCE under the same conditions. The experimental results showed that the biosensor we prepared has a good response to them. The linear range and detection limit of the compounds are shown in Table 1.
Table 1

Linear range and detection limit

Compound

Linear range (M)

Detection limit (M)

Correlation coefficient

Phenol

4.0 × 10−7 to 1.0 × 10−5

2.0 × 10−7

0.9995

Hydroquinone

2.0 × 10−7 to 6.0 × 10−5

2.0 × 10−7

0.9989

Catechol

2.0 × 10−7 to 1.0 × 10−5

2.0 × 10−7

0.9990

Bisphenol A

2.0 × 10−6 to 1.0 × 10−4

5.0 × 10−7

0.9982

Reproducibility, stability and interference

The repeatability of the current response of one enzyme electrode to 50 μM phenol was examined. The relative standard deviation (RSD) was 3.4% for eight successive assays. The electrode-to-electrode reproducibility was determined from the response to 50 μM phenol at five different enzyme electrodes, an acceptable reproducibility was obtained with a variation coefficient of 4.0%.

The enzyme electrode was stored in a dry state at 4 °C in a refrigerator when not in use. The enzyme electrode retained 72% of its original response after 1 month testing.

In order to evaluate the selectivity of the biosensor, the influence of some possible interfering substances was examined in phosphate buffer solution (pH 7.5) containing 20 μM phenol. The results suggested that 100-fold concentration of K+, Ca2+, Mg2+, Zn2+, Pb2+, Cu2+, Cl, Br, I, SO42−, PO43−, CO32−, NO3, 100-fold concentration of glucose, 50-fold concentration of L-cysteine and uric acid, 20-fold concentration of aminoacetic acid, 5-fold concentration of benzoic acid did not interfere with the determination of phenol.

Analytical applications

To study the real feasibility of the biosensor, we checked the biosensor in determination of phenol in real water samples (obtained from Beijing south moat). No special sample pretreatment was required, after the water samples were filtered with a 0.45 μm filter to eliminate particulate matters, the determination can be proceeded. The results were shown in Table 2. In addition, standard addition method was adopted to estimate the accuracy of the method, and the results were also presented in Table 2. It can be seen that the results were satisfactory.
Table 2

Recovery of tyrosinase biosensor (n = 6)

Sample

Found (M)

Phenol added (M)

Found (M)

Recovery (%)

1

3.98 × 10−7

4.00 × 10−7

4.14 × 10−7

103.5

2

3.70 × 10−7

4.00 × 10−7

4.22 × 10−7

105.5

3

3.55 × 10−7

4.00 × 10−7

3.77 × 10−7

94.3

Comparison of the biosensor for determination of phenol with others

Table 3 summarises some characteristics of recent phenol biosensors compared to our biosensor. As it can be observed, the analytical range and detection limit values of the biosensors fabricated by us are better than many methods reported in recent publications. The immobilization matrix is easy to prepare and cheaper than other materials like polyacrylamicrogels microgels [12], sol-gel silicate/Nafion composite [29] and PDDA [42]. The biosensor exhibited better long-term stability than that reported in references [24, 37, 43]. Moreover, the method can be applied to the determination of phenol in real water samples.
Table 3

Comparison of the biosensor fabricated by this paper for determination of phenol with others (GCE, glassy carbon electrode; CPE, carbon paste electrode; MWNTs, multiwalled carbon nanotubes; PDDA, poly(dimethyldiallylammonium) chloride; Tyr, tyrosinase)

Electrode

Modification

Stability

Immobilization technique

LOD(μM)

Analytical range(M)

Refs.

CPE

MgFe2O4-SiO2

70% after 30 days

Covalently immobilized to MgFe2O4-SiO2

0.60

1.0 × 10−6–2.5 × 10−4

[11]

GCE

Polyacrylamicrogels microgels

100% after 20 days

Cross-link

1.40

5.0 × 10−6–2.2 × 10−5

[12]

Carbon electrode

Alumina sol–gel

Loss 100% after 3 weeks

Mix Tyr and sol–gel solution

0.30

5.0 × 10−7–3.0 × 10−5

[24]

GCE

Sol–gel silicate/Nafion composite

74% after 2 weeks

MixTyr and sol–gel silicate/Nafion

1.00

5.0 × 10−6–1.0 × 10−4

[29]

GCE

MWNTs, PDDA

Layer by layer self-assembled

0.66

2.0 × 10−6–1.0 × 10−4

[42]

GCE

MWNTs, Nafion

Mix Tyr, MWNTs and Nation

0.13

1.0 × 10−6–1.9 × 10−5

[44]

GCE

Palygorskite

80% after 60 days

Mix Tyr and palygorskite

5.0 × 10−8–1.0 × 10−4

[45]

GCE

MWNTs

72% after 30 days

Cross-linking with glutaraldehyde

0.20

4.0 × 10−7–1.0 × 10−5

This work

Conclusions

We describe an amperometric biosensor here which was fabricated based on tyrosinase immobilized onto MWNTs film for the determination of phenolic compounds. The MWNTs film enhanced the direct electron transfer rate of benzoquinone produced by enzymatic reaction at the electrode. Compared with those biosensors reported in other literatures, the biosensors reported in this work possess main merits are simple fabrication method, good performances in terms of response rate, high sensitivity and operational stability.

Notes

Acknowledgements

This work was supported by the National Natural Science Foundation of China (No. 20247002), Beijing Natural Science Foundation (No.8102009), the Beijing Municipal Education Commission Scientific Technological Project Foundation (No. KZ201110005006) and Beijing Municipal institution of higher learning academic innovating group projects (No. PHR 201007105).

References

  1. 1.
    Li YF, Liu ZM, Liu YL, Yang YH, Shen GL, Yu RQ (2006) A mediator-free phenol biosensor based on immobilizing tyrosinase to ZnO nanoparticles. Anal Biochem 349:33–40CrossRefGoogle Scholar
  2. 2.
    Rajesh TW, Kaneto K (2004) Amperometric phenol biosensor based on covalent immobilization of tyrosinase onto an electrochemically prepared novel copolymer poly(N-3-aminopropyl pyrrole-co-pyrrole) film. Sens Actuators B Chem 102:271–277CrossRefGoogle Scholar
  3. 3.
    Lupetti KO, Rocha FRP, Fatibello-Filho O (2004) An improved flow system for phenols determination exploiting multicommutation and long pathlength spectrophotometry. Talanta 62:463–467CrossRefGoogle Scholar
  4. 4.
    Vanbeneden N, Delvaux F, Delvaux FR (2006) Determination of hydroxycinnamic acids and volatile phenols in wort and beer by isocratic high-performance liquid chromatography using electrochemical detection. J Chromatogr A 1136:237–242CrossRefGoogle Scholar
  5. 5.
    Portaccio M, Tuoro DD, Arduini F, Lepore M, Mita DG, Diano N, Mita L, Moscone D (2010) A thionine-modified carbon paste amperometric biosensor for catechol and bisphenol A determination. Biosens Bioelectron 25:2003–2008CrossRefGoogle Scholar
  6. 6.
    Mita DG, Attanasio A, Arduini F, Diano N, Grano V, Bencivenga U, Rossi S, Amine A, Moscone D (2007) Enzymatic determination of BPA by means of tyrosinase immobilized on different carbon carriers. Biosens Bioelectron 23:60–65CrossRefGoogle Scholar
  7. 7.
    Zhang J, Lei JP, Liu YY, Zhao JW, Ju HG (2009) Highly sensitive amperometric biosensors for phenols based on polyaniline–ionic liquid–carbon nanofiber composite. Biosens Bioelectron 24:1858–1863CrossRefGoogle Scholar
  8. 8.
    Notsu H, Tatsuma T, Fujishima A (2002) Tyrosinase-modified boron-doped diamond electrodes for the determination of phenol derivatives. J Electroanal Chem 523:86–92CrossRefGoogle Scholar
  9. 9.
    Abdullah J, Ahmada M, Heng LY, Karuppiah N, Sidek H (2006) Chitosan-based tyrosinase optical phenol biosensor employing hybrid nafion/sol–gel silicate for MBTH immobilization. Talanta 70:527–532CrossRefGoogle Scholar
  10. 10.
    Cosnier S, Ionescu RE, Holzinger M (2008) Aqueous dispersions of SWCNTs using pyrrolic surfactants for the electro-generation of homogeneous nanotube composites. Application to the design of an amperometric biosensor. J Mater Chem 18:5129–5133CrossRefGoogle Scholar
  11. 11.
    Liu ZM, Liu YL, Yang HF, Yang Y, Shen GL, Yu RQ (2005) A phenol biosensor based on immobilizing tyrosinase to modified core-shell magnetic nanoparticles supported at a carbon paste electrode. Anal Chim Acta 533:3–9CrossRefGoogle Scholar
  12. 12.
    Hervás Pérez JP, Sánchez-Paniagua López M, López-Cabarcos E, López-Ruiz B (2006) Amperometric tyrosinase biosensor based on polyacrylamide microgels. Biosens Bioelectron 22:429–439CrossRefGoogle Scholar
  13. 13.
    Shan D, Zhu MJ, Han E, Xue HG, Cosnier S (2007) Calcium carbonate nanoparticles: a host matrix for the construction of highly sensitive amperometric phenol biosensor. Biosens Bioelectron 23:648–654CrossRefGoogle Scholar
  14. 14.
    Alkasir RSJ, Ganesana M, Won YH, Stanciu L, Andreescu S (2010) Enzyme functionalized nanoparticles for electrochemical biosensors: a comparative study with applications for the detection of bisphenol A. Biosens Bioelectron 26:43–49CrossRefGoogle Scholar
  15. 15.
    Liu SQ, Yu JH, Ju HX (2003) Renewable phenol biosensor based on a tyrosinase-colloidal gold modified carbon paste electrode. J Electroanal Chem 540:61–67CrossRefGoogle Scholar
  16. 16.
    Xia W, Li YY, Wan YJ, Chen T, Wei J, Lin Y, Xu SQ (2010) Electrochemical biosensor for estrogenic substance using lipid bilayers modified by Au nanoparticles. Biosens Bioelectron 25:2253–2258CrossRefGoogle Scholar
  17. 17.
    Wang P, Liu M, Kan JQ (2009) Amperometric phenol biosensor based on polyaniline. Sens Actuators B Chem 140:577–584CrossRefGoogle Scholar
  18. 18.
    Wang G, Xu JJ, Ye LH, Zhu JJ, Chen HY (2002) Highly sensitive sensors based on the immobilization of tyrosinase in chitosan. Bioelectrochemistry 57:33–38CrossRefGoogle Scholar
  19. 19.
    Dempsey E, Diamond D, Collier A (2004) Development of a biosensor for endocrine disrupting compounds based on tyrosinase entrapped within a poly(thionine) film. Biosens Bioelectron 20:367–377CrossRefGoogle Scholar
  20. 20.
    Xue HG, Shen ZQ (2002) A highly stable biosensor for phenols prepared by immobilizing polyphenol oxidase into polyaniline–polyacrylonitrile composite matrix. Talanta 57:289–295CrossRefGoogle Scholar
  21. 21.
    Yildiz HB, Castillo J, Guschin DA, Toppare L, Schuhmann W (2007) Phenol biosensor based on electrochemically controlled integration of tyrosinase in a redox polymer. Microchim Acta 159:27–34CrossRefGoogle Scholar
  22. 22.
    Zhang T, Tian B, Kong J, Yang P, Liu B (2003) A sensitive mediator-free tyrosinase biosensor based on an inorganic–organic hybrid titania sol–gel matrix. Anal Chim Acta 489:199–206CrossRefGoogle Scholar
  23. 23.
    Yu JH, Liu SQ, Ju HX (2003) Mediator-free phenol sensor based on titania sol–gel encapsulation matrix for immobilization of tyrosinase by a vapor deposition method. Biosens Bioelectron 19:509–514CrossRefGoogle Scholar
  24. 24.
    Zejli H, Hidalgo-Hidalgo de Cisneros JL, Naranjo-Rodriguez I, Liu B, Temsamani KR, Marty JL (2008) Phenol biosensor based on Sonogel-Carbon transducer with tyrosinase alumina sol–gel immobilization. Anal Chim Acta 612:198–203CrossRefGoogle Scholar
  25. 25.
    Campuzano S, Serra B, Pedrero M, Manuel J, de Villena F, Pingarrón JM (2003) Amperometric flow-injection determination of phenolic compounds at self-assembled monolayer-based tyrosinase biosensors. Anal Chim Acta 494:187–197CrossRefGoogle Scholar
  26. 26.
    Tatsuma T, Sato T (2004) Self-wiring from tyrosinase to an electrode with redox polymers. J Electroanal Chem 572:15–19CrossRefGoogle Scholar
  27. 27.
    Anh TM, Dzyadevych SV, Soldatkin AP, Chien ND, Jaffrezic-Renault N, Chovelon JM (2002) Development of tyrosinase biosensor based on pH-sensitive field-effect transistors for phenols determination in water solutions. Talanta 56:627–634CrossRefGoogle Scholar
  28. 28.
    Rogers KR, Becker JY, Cembrano J (2000) Improved selective electrocatalytic oxidation of phenols by tyrosinase-based carbon paste electrode biosensor. Electrochim Acta 45:4373–4379CrossRefGoogle Scholar
  29. 29.
    Kim MA, Lee WY (2003) Amperometric phenol biosensor based on sol–gel silicate/Nafion composite film. Anal Chim Acta 479:143–150CrossRefGoogle Scholar
  30. 30.
    Li C (2007) Voltammetric determination of 2-chlorophenol using a glassy carbon electrode coated with multi-wall carbon nanotube-dicetyl phosphate film. Microchim Acta 157:21–26CrossRefGoogle Scholar
  31. 31.
    Zheng YQ, Yang CZ, Pu WH, Zhang JD (2009) Carbon nanotube-based DNA biosensor for monitoring phenolic pollutants. Microchim Acta 166:21–26CrossRefGoogle Scholar
  32. 32.
    Zhang H, Zhao JS, Liu HT, Liu RM, Wang HS, Liu JF (2010) Electrochemical determination of diphenols and their mixtures at the multiwall carbon nanotubes/poly (3-methylthiophene) modified glassy carbon electrode. Microchim Acta 169:277–282CrossRefGoogle Scholar
  33. 33.
    Li JH, Kuang DZ, Feng YL, Zhang FX, Liu MQ (2011) Voltammetric determination of bisphenol A in food package by a glassy carbon electrode modified with carboxylated multi-walled carbon nanotubes. Microchim Acta 172:379–386CrossRefGoogle Scholar
  34. 34.
    Jiang CM, Chen H, Yu C, Zhang S, Liu BH, Kong JL (2009) Preparation of the Pt nanoparticles decorated poly(N-acetylaniline)/MWNTs nanocomposite and its electrocatalytic oxidation toward formaldehyde. Electrochim Acta 54:1134–1140CrossRefGoogle Scholar
  35. 35.
    Li J, Jessica EK, Alan MC, Hua C, Tee NH, Qi Y, Wendy F, Jie H, Meyyapan M (2005) Inlaid multi-walled carbon nanotube nanoelectrode arrays for electroanalysis. Electroanalysis 17:15–27CrossRefGoogle Scholar
  36. 36.
    Huang JL, Tsai YC (2009) Direct electrochemistry and biosensing of hydrogen peroxide of horseradish peroxidase immobilized at multiwalled carbon nanotube/alumina-coated silica nanocomposite modified glassy carbon electrode. Sens Actuators B Chem 140:267–272CrossRefGoogle Scholar
  37. 37.
    Korkut S, Keskinler B, Erhan E (2008) An amperometric biosensor based on multiwalled carbon nanotube-poly(pyrrole)-horseradish peroxidase nanobiocomposite film for determination of phenol derivatives. Talanta 76:1147–1152CrossRefGoogle Scholar
  38. 38.
    Yin HS, Zhou YL, Xu J, Ai SY, Cui L, Zhu LS (2010) Amperometric biosensor based on tyrosinase immobilized onto multiwalled carbon nanotubes-cobalt phthalocyanine-silk fibroin film and its application to determine bisphenol A. Anal Chim Acta 659:144–150CrossRefGoogle Scholar
  39. 39.
    Sanz VC, Mena M, González-Cortés A, Yanez-Sedeno P, Pingarrón J (2005) Development of a tyrosinase biosensor based on gold nanoparticles-modified glassy carbon electrodes: application to the measurement of a bioelectrochemical polyphenols index in wines. Anal Chim Acta 528:1–8CrossRefGoogle Scholar
  40. 40.
    Vedrine C, Fabiano S, Tran-Minh C (2003) Amperometric tyrosinase based biosensor using an electrogenerated polythiophene film as an entrapment support. Talanta 59:535–544CrossRefGoogle Scholar
  41. 41.
    Wang J, Liu J, Cepra G (1997) Thermal stabilization of enzymes immobilized within carbon paste electrodes. Anal Chem 69:3124–3127CrossRefGoogle Scholar
  42. 42.
    Kong LM, Huang SS, Yue ZL, Peng B, Li MY, Zhang J (2009) Sensitive mediator-free tyrosinase biosensor for the determination of 2,4-dichlorophenol. Microchim Acta 165:203–209CrossRefGoogle Scholar
  43. 43.
    Diaconu M, Litescu SC, Radu GL (2011) Bienzymatic sensor based on the use of redox enzymes and chitosan–MWCNT nanocomposite. Evaluation of totalphenolic content in plant extracts. Microchim Acta 172:177–184CrossRefGoogle Scholar
  44. 44.
    Tsai YC, Chiu CC (2007) Amperometric biosensors based on multiwalled carbon nanotube—Nafion-tyrosinase nanobiocomposites for the determination of phenolic compounds. Sens Actuators B 125:10–16CrossRefGoogle Scholar
  45. 45.
    Chen J, Jin YL (2010) Sensitive phenol determination based on co-modifying tyrosinase and palygorskite on glassy carbon electrode. Microchim Acta 169:249–254CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Jing Ren
    • 1
  • Tian-Fang Kang
    • 1
  • Rui Xue
    • 1
  • Chao-Nan Ge
    • 1
  • Shui-Yuan Cheng
    • 1
  1. 1.College of Environmental and Energy EngineeringBeijing University of TechnologyBeijingChina

Personalised recommendations