Alcohol quantification: recent insights into amperometric enzyme biosensors

Figures & data

Table 1. Different methods other than enzymatic approaches used for the measurement of ethanol in various types of samples.

Sr. No.	Technique	Sample	Linear range	Detection limit	References
1.	GC/FID, direct injection	Biological fluids	0.1–5000 mg/l	0.1 ppm	[5]
2.	GC/FID, headspace injection	Blood	–	0.3 ppm	[6]
3.	FT-NIR spectrometry	Fuel and alcoholic beverages	–	0.05% (w/w)	[7]
4.	Pulse amperometry, Pt electrodes	Alcoholic beverages	0.02–1700 mM	0.02 mM	[8]
5.	NIR spectrophotometry, flow injection	Liquor	200–500 g/l	10 g/l	[9]
6.	HPLC/FTIR	Wine	–	1–10 mg/ml	[10]
7.	Photometry, Cr(VI) Cr(III) at 600 nm	Spirits	5–40%	5%	[11]

FID: flame-ionization detection; FT: Fourier transform; GC: gas chromatography; HPLC: high performance liquid chromatography; IP: infrared; NIR: near infrared.

Figure 1. Generalized schematic showing the key components of an enzymatic biosensor.

Figure 2. Schematics showing coupling of two enzymes for the detection of ethanol.

Table 2. AOX-based biosensors.

Sr. No.	Mode of detection	Transducer	Enzyme immobilization method	Minimum detection limit	Linearity	Response time	References
1.	Amperometric	Nafion/HRP-SWCNT/MWCNT-ZnO electrode	Crosslinking	10 μM	10 μM–750 μm	10 s	[24]
2.	Amperometric	Carbon film/PNR/AOX electrode	Crosslinking	29.7 ± 1.5 μM	–	–	[25]
3.	Amperometric	Graphite epoxy composite electrode (GECE)	Entrapment	–	2.5–25 μM	70 s	[20]
4.	Conductometric	poly(vinyl alcohol) membrane/interdigitated microelectrode	Entrapment	1 μM	70 μM	–	[26]
5.	Amperometric	Gas-diffusion working electrode	Crosslinking	9.9 μM	0.10–30 mM	69 s	[27]
6.	Amperometric	Eggshell membrane electrode	Entrapment	30 μM	60 μM–0.80 mM	60 s	[28]
7.	Amperometric	Multi-walled carbon nanotube (MWCNT) modified glassy carbon electrode	Layer-by-layer technique	2.3 × 10^−6 mol L^−1	5.0 × 10^−6–30 × 10^−4 mol L^−1	–	[29]
8.	Amperometric	Au/MWCNT/Nf/AOX/PEI bioelectrodes	Crosslinking	5 μM	8 μM–42 μM	55 s	[30]
9.	Amperometric	Graphite powder/an epoxy resin/AOX bicomposite electrode	Entrapment	–	0.5–29 mg/ml	–	[31]
10.	Amperometric	PEDOT/TBTD/AOX electrode	Crosslinking	203.70 nA/mM	0.02–1.7 mM	–	[32]

AOX: Alcohol oxidase; HRP: horseradish peroxidase; MWCNT: multiwall carbon nanotube; Nf: Nafion; PEI: polyethylenimine; PEDOT: 3, 4-ethylenedioxythiophene; SWCNT: single wall carbon nanotube; TBTD: 4, 7-dithien-2-yl-2, 1, 3-benzothiadiazole.

Figure 3. ADH catalyzed reaction for the detection of ethanol at biosensor surface.

Table 3. ADH-based biosensors.

Sr. No.	Mode of detection	Transducer	Enzyme immobilization method	Minimum detection limit	Linearity	Respone time	References
1.	Amperometric	GNPs–MWCNT–Nafion nanocomposite film modified screen printed electrode	Covalent bonding	5.0 × 10^−5 mol/L	2.0 × 10^−4–25 × 10^−3 mol/L	–	[40]
2.	Amperometric	Multi-wall carbon nanotube (MWCT)/Meldola's blue electrode	Crosslinking	–	0.05–10 mmol L^−1	–	[2]
3.	Amperometric	Poly(vinyl alcohol)–multiwalled carbon nanotube (PVA–MWCNT) composite	Physical adsorption	–	up to 1.5 mM	8s	[41]
4.	Amperometric	Silver wire electrode	Covalent bonding	less than 0.1 mM	0–12.5 mM	50 s–4 min	[42]
5.	Voltametric	Cellulose acetate (CA)/toluidine blue O (TBO)/glassy electrode	Crosslinking	5.0 × 10^−6 M	1 × 10^−5 M–4 × 10^−4 M	–	[43]
6.	Amperometric	Os-complex-modified poly(vinylimidazole) redox polymer	Crosslinking	1 μM	–	–	[35]
7.	Amperometric	MWCNTS/AuNPs/PNR/ADH modified screen printed carbon electrode	Crosslinking	96.1 μM	320.2 μM–1000 μM	–	[44]
8.	Amperometric	MDB/GMCs/ADH/NAD^+ modified screen printed electrode	Crosslinking	80 μM	0.5–15 mM		[45]
9.	Optical	RuSiNPs/ADH modified glassy carbon electrode	Crosslinking	5.0 × 10^−8 M	10^−7–10^−2 M		[46]
10.	Amperometric	GC/SWCNT-Polytry(oxidized)/ADH/NAF	Crosslinking	7.9 nM	–	–	[47]
11.	Amperometric	CNTP/Meldola Blue/ADH/NAD^+	Crosslinking	1 × 10^−7 mol/L	1 × 10^−7–4 × 10^−6 mol/L	–	[48]
12.	Amperometric	MWCNT/ADH/NAD^+/Teflon matrix	Crosslinking	0.1 mM	–	–	[49]

ADH: alcohol dehydrogenase; GC: glassy carbon; GMC: graphitized mesoporous carbon; GNP: Gold nanoparticles; MDB: Meldola's blue; MWCNT: multi wall carbon-nanotube; NAF: Nafion; RuSiNPS: Ru(bpy)(3)(2+)-doped silica nanoparticles; SWCNT: single wall carbon nanotube.

Figure 4. ADH catalyzed reaction for the detection of ethanol at biosensor surface.

Figure 5. Scheme showing the reaction sequence within the enzyme electrode using a mediator.

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