Fresh broad (Vicia faba) tissue homogenate-based biosensor for determination of phenolic compounds

Figures & data

Schema 1. Concept of process.

Process steps	Process
1	200 mg of fresh broads were weighed and homogenized with 500 μL working phosphate buffer (66 mM, pH 7). The mixture of 270 μl fresh broad homogenate and 10 mg gelatin was incubated at 38°C for 15 min to dissolve gelatin.
2	30 μL Gelatin-fresh broad tissue homogenate was dispersed over the cleaned GCE surface.
3	It was allowed to dry at 4 •C for 30 min.
4	For cross linking with glutaraldehyde, the probe carrying bioactive layer was immersed into 1.25% (v/v) glutaraldehyde solution and was allowed to stay for 5 min.
5	The biosensor was washed with distilled water to remove excess glutaraldehyde and it was ready to use.

Figure 1. Effect of the fresh broad amount on the biosensor response.

Figure 2. Effect of the gelatin amount on the biosensor response.

Figure 3. Effect of the glutaraldehyde percent on the biosensor response.

Figure 4. Effect of the pH on the biosensor activity.

short-legendFigure 5.

Figure 6. Calibration curve for catechol.

Table I. Inhibition studies.

Substances^a,d	Inhıbition (%)^e	Inhıbition (%)^f
Cysteine	12.9	10.7
Hg^2+b	21.2	18.7
Sulfide	9.3	5.2
Bisulfide	9.5	6.2
Cu^2+c	1.3	0.9
Zn^2+c	1.1	1.0
Benzoic acid	4.3	4.2
Thiourea	3.5	3.0

^aConcentrations of substances tested and catechol used as a substrate were 100 and 200 μM, respectively.

^bHgCl₂ was used for Hg²⁺ ions.

^cCuSO₄ and ZnSO₄ were used for Cu²⁺ and Zn²⁺ ions.

^dMean of three determination.

^eInhibition values for banana peel-based biosensor (our previous study (Ozcan and Sagıroglu 2010)).

^fInhibition values for fresh broad-based biosensor.

Table II. Results obtained from the measurements of some phenolic compounds.

Phenolic compound	Activity(%)	Linear Range (μM)	R^2	Y
Catechol	100.00	5–60	0.999	108.9x + 15.02
Pyrogallol	87.63	10–60	0.996	209.1x + 8.71
P-Cresol	136.08	10–50	0.990	224x + 16.06
Hydroqinone	121.13	10–60	0.997	157.2x + 16.96
phenol	68.56	10–40	0.996	80x + 10.05
L-Dopa	68.04	20–60	0.997	105x + 8.94
Gallic Acid	65.98	10–70	0.998	122.3x + 7.97
Caffeic Acid	133.50	10–60	0.993	120.4x + 20.82
Rutin	59.28	10–40	0.995	70x + 8.65
Cinnamic Acid	110.10	10–50	0.998	172x + 25.76
Resorsine	69.59	10–70	0.999	242.3x + 3.75
Quersetine	65.46	20–60	0.988	31x + 11.4
Ascorbic Acid	98	10–60	0.998	241.5x + 5.63

Table III. Comparison of the performance of banana peel and fresh broad biosensor for determination of different phenolic compounds.

Phenolic compound	Activity(%)^a	Activity(%)^b	Linear Range (μM)^a	Linear Range (μM)^b
Catechol	100	100.00	20–80	5–60
Pyrogallol	117.7	87.63	10–60	10–60
P-Cresol	96	136.08	10–50	10–50
Hydroqinone	106.7	121.13	10–70	10–60
phenol	63.6	68.56	10–40	10–40
L-Dopa	50	68.04	20–60	20–60
Gallic Acid	83.3	65.98	10–80	10–70
Caffeic Acid	139.9	133.50	20–70	10–60
Rutin	55.6	59.28	20–50	10–40
Cinnamic Acid	108.1	110.10	10–60	10–50
Resorsine	65.1	69.59	10–40	10–70
Quersetine	55.1	65.46	20–60	20–60
Ascorbic Acid	96	98	20–80	10–60

^aBanana peel- based biosensor (Ozcan and Sagıroglu 2010).

^bFresh broad-based biosensor.

Table IV. Phenolic compound measurements in some drink samples.

Substances^a	Included amount (μM)	Calculated amount of calibration curve (μM)	Recovery (%)
Apple Juice (Pınar %100)	22 50	22.08 ± 0.46 50.68 ± 0.82	100.36 101.36
CocaCola	32 54	32.32 ± 0.50 54.18 ± 0.77	101.00 100.34
Fanta	27 50	27.36 ± 0.65 70.09 ± 0.46	101.34 100.18
Grape Juice (Tamek %100)	22 54	22.58 ± 0.77 54.54 ± 0.53	104.55 101.10
Beer (Efes Pilsen)	12 50	11.93 ± 0.41 49.58 ± 0.81	99.42 99.18
White Wine (Guzel Marmara)	16 46	15.98 ± 0.49 46.10 ± 0.82	99.87 100.22

^aMean of five determimation.

Ozcan HM, Sagiroglu A. 2010. A novel amperometric biosensor based on banana peel (Musa cavendish) tissue homogenate for determination of phenolic compounds. Artif Cells Blood Substitutes Biotechnol. 38:208–214.

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