Recent trends and perspectives in enzyme based biosensor development for the screening of triglycerides: a comprehensive review

Figures & data

Figure 1 Reference range of triglycerides and various disorders associated with high level of TGs in human body.

Table 1. Various conventional methods for triglyceride determination with their merits and demerits.

Conventional Methods	Merits	Demerits	References
1. Chemical methods
Titrimetric method	Reproducible results, easy to use	Non-specific, interference by other biological compounds	[5]
Micro method	Good linearity range	Expensive, requirement of many devices for analysis, less reliable	[8]
Autoanalyzer method	Speedy results, high reproducibility, low variation in results	Necessity of trained and competent personnel, poorly used in clinical laboratories	[15,16]
2. Chromatographic methods
HPLC method	Faster, cost-effective, sensitive	Time consuming, costly equipments, requirement of expertise handling	[17]
Thin layer chromatographic method	Nanogram concentrations of triglyceride could be measured, high sensitivity	Difficulty in reproducing TLC results, faint and uneven spotting	[18]
Capillary ion-column gas chromatography method	Cost-effective, high signal to noise ratio, fast response time	Only compounds having high vapor pressures can be analyzed	[19]
3. Mass fragmentographic method	Sensitive	Requirement of expensive equipments	[20]

HPLC: High performance liquid chromatography.

Figure 2 Schematic diagram showing basic principles of the biosensors.

Figure 3 Schematic representation of electrochemical reactions involved in response measurement of enzyme based amperometric TG biosensor.

Table 2. Membrane and DO metric based amperometric TG biosensors and comparison of their analytical properties.

Mode of Detection	Transducer	Immobilization Method	Linearity	Minimum detection limit	Storage stability	Responsetime	References
Amperometric	LIP/GK/GPO/HRP/PVC	Crosslinking	0.56–2.25 mM	0.11 mM	40 days	30 s	[15]
Amperometric	LIP/GK/GPO/GCE/ gelatin membrane	Crosslinking	0.14–1.125 mM	0.14 mM	30 days	NR	[28]
Amperometric	LIP/GK/GPO/Egg shell membrane/Au	Physical adsorption	0.56–2.25 mM	0.28 mM	200 times used within 70 days	10 s	[29]
Amperometric	LIP/GK/GPO/HRP/PVC	Crosslinking	0.1–15 mM	0.1 mM	50% decay after 200 uses	NR	[30]
Amperometric	Diaphorase/GDH/ Collagen membrane	Physical adsorption	0.11–5.6 mM	0.1 mM	27 days	900 s	[31]
Amperometric	LIP/GK/GPO/POD/glass beads	Crosslinking	0.20–1.38 mM	0.20 mM	50% decay after 150 uses	1200 s	[16]
DO	LIP/GK/GPO/CA	Crosslinking	0.2–3.5 mM	0.2 mM	150 times used within 25 days	40 s	[16]
DO	LIP/GDH/Oxygen electrode	NA	0.05–0.3 mM	0.05 mM	NR	300 s	[32]
DO	LIP/GK/GPO/HRP/PVA	Crosslinking	0.56–2.25 mM	0.21 mM	50% decay in 50 days	2 s	[17]
DO	LIP/GK/GPO/PUM	Physical adsorption	0.005–1 mM	0.005 mM	NR	90 s	[33]
DO	LIP/GK/GPO/PVC	Crosslinking	5–20 mM	0.35 mM	NR	10 s	[34]

LIP: Lipase; GK: glycerol kinase; GPO: glycerol-3-phosphate oxidase; GDH: glycerol dehydrogenase; HRP: horse radish peroxidase; GCE: glassy carbon electrode; PVC: polyvinyl chloride; PUM: poly urethane membrane; PVA: polyvinyl alcohol.

Table 3. Potentiometric /ion selective electrode based TG biosensors and comparison of their analytical properties.

Mode of detection	Transducer	Immobilization method	Linearity	Minimum detection limit	Storage stability	Response time	References
Potentiometric	Psi based electrode	Physical adsorption	5.9–21 mM	NR	180 − days	900 s	[38]
Potentiometric	Mesoporous Psi matrix	Physical adsorption	<5.6 mM	NR	17 hours working time within 60 days	NR	[39]
Potentiometric	EISCAP	NR	5–15 mM	5 mM	NR	NR	[40]
Potentiometric	Crystalline silica based miniaturized EISCAP	Covalent coupling	5 mM	NR	NR	30 s	[41]
Potentiometric	Microcantilever sensor	NR	0.024–0.48 mM	0.01 mM	NR	1200 s	[42]
Potentiometric	Silicon nitride layer of EISCAP	Covalent coupling	1–7 mM	5 mM	NR	2700 s	[42]
Potentiometric	EISCAP	NR	0.1–18 mM	0.1 mM	NR	1800 s	[43]
Potentiometric	Miniturized EISCAP	Covalent coupling	0.56–1.68 mM	0.56 mM	NR	300 s	[44]
Potentiometric	LIP/Pt wire (pH-FET)	Physical adsorption	0.6–3 mM	0.00009 mM	44 days	120 s	[45]
Potentiometric	Gate surface of a FET	Physical adsorption	5–30 mM	5 mM	45% decay after 14 days	<900 s	[46]

FET: Field effect transisitors; Psi: porous silicon; EISCAP: electrolyte insulator semiconductor capacitors.

Figure 4 Diagrammatic representation of TG biosensor based on porous silicon.

Figure 5 Schematic diagram of enzyme immobilized mini-EISCAP.

Table 4. Conducting polymer, nanostructured metal oxides (NMOs) and ENPs based TG biosensors and comparison of their analytical properties.

Mode of detection	Transducer	Immobilization method	Linearity	Minimum detection limit	Storage stability	Response time	References
Amperometric	LIP/GDH/PANI/SWCNT/ITO	Covalent coupling	0.56–4.52 mM	0.56 mM	91 days	12 s	[50]
Amperometric	LIP/GK/GPO	Covalent coupling	0.17–2.26 mM	0.17 mM	NR	NR	[51]
Amperometric	LIP/GK/GDH/IrNPs/CPE	Covalent coupling	0–10 mM	0.1 mM	NR	NR	[55]
Amperometric	LIP/Nano-CeO2/ITO	Covalent coupling	0.56–5.65 mM	0.37 mM	84 days	NR	[56]
Amperometric	LIP/GK/GPO/CHIT/ZnONPs	Covalent coupling	0.56 7.35 mM	0.23 mM	50% decay in 210 days	6 s	[57]
Amperometric	LIP/NPG/GCE	Covalent coupling	0.56–7.91 mM	0.23 mM	50% decay after 100 uses in 210 days	4 s	[58]
Amperometric	LIP/GK/GPO/PtNPs/PPY/ITO	Covalent coupling	0.56–5.65 mM	0.28 mM	270 days	NR	[30]
Amperometric	NiONPs-CHIT/ZnO-ZnHCF/LIP/GK/GPO/Au	Covalent coupling	0.56–7.91 mM	0.11 mM	50% decay in 180 days	4 s	[59]
Amperometric	LIP/GK/GPO/Inkjet-printed Au/PEDOT-PSS/SPCE	Covalent coupling	0–6 mM	0.09 mM	40% decay decay in 30 days	30 s	[20]
Amperometric	LIP/NanoCeO2-SiO2/ITO	Covalent coupling	0.565–4.52 mM	0.565 mM	NR	12 s	[60]
Amperometric	LIP/GK/GPO/MNP-CHIT/ZnO-ZnHCF/GCE	Covalent coupling	0.56–2.83 mM	0.03 mM	NR	NR	[61]
Amperometric	LIP/GK/GPO/MNP-CHIT/ZnO-ZnHCF/GCE	Covalent coupling	0.11–11.3 mM	0.11 mM	NR	NR	[62]
Amperometric	LIP/GDH/TB/ERGO/ITO	Covalent coupling	0.56–3.39 mM	0.56 mM	60 days	NR	[63]
Amperometric	LIP/GK/GPO/CHIT-nano-ZrO2/ITO	Covalent coupling	0.0001–0.05 mM	0.000155 mM	85% decay in 77 days	45 s	[64]
Amperometric	LIPNPs/GKNPs/GPONPs/Au	Covalent coupling	0.11–5.65 mM	0.001 mM	90 days	5 s	[65]
Amperometric	LIPNPs/GKNPs/GPONPs/PGE	Physical adsorption	0.1–45	10^-7 mM	20% decay in 240 days	2.5 s	[66,67]

LIP: lipase; GK: glycerol kinase; GPO: glycerol-3-phosphate oxidase; ITO: indium-tin- oxide; GDH: glycerol dehydrogenase; CHIT: chitosan; GCE: glassy carbon electrode; CeO₂: cerium oxide; ZnHCF: zinc hexacyanoferrate; TB: tributyrin; IrNPs: iridium nanoparticles; ERGO: electrochemically reduced graphene oxide; ZnO: zinc oxide; ZnONPs: zinc oxide nanoparticles; NPG: nanoporous gold; PPY: polypyrrole.

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