Simple and Cost-effective Enzymatic Detection of Cholesterol Using Flow Injection Analysis

Overview

Journal Anal Sci

Publisher Springer

Specialty Chemistry

Date 2020 Sep 10

PMID 32908068

Citations 2

Authors

Murugan Thiruppathi

Ching-Ying Tsai

Tzu-Wen Wang

Yu Tsao

Tsung-Hung Wu

Ja-An Annie Ho

Affiliations

Soon will be listed here.

Abstract

A flow-injection analytical (FIA) system was developed for the determination of cholesterol concentrations based on enzymatic reactions that occurred in a cholesterol oxidase (CHO)-immobilized, fused-silica capillary followed by electrochemical detection. The production of hydrogen peroxide from cholesterol in an enzymatic reaction catalyzed by CHO was subsequently oxidized electrochemically at an electrode. Our FlA system demonstrated its cost-effectiveness and utility at an applied potential of 0.6 V (vs. Ag/AgCl), a flow rate of 100 μL/min and, under optimal conditions, the resulting signal demonstrated a linear dynamic range from 50 μM to 1.0 mM with a limit of detection (LOD) of 12.4 μM, limit of quantification (LOQ) of 44.9 μM, and the coefficient of variation of 5.17%. In addition, validation of our proposed system using a reference HDL-cholesterol kit used for clinical diagnosis suggested our FIA system was comparable to commercial kits for the determination of the cholesterol incorporation amount in various aqueous liposomal suspensions. These good analytical features achieved by FIA could make the implementation of this methodology possible for on-line monitoring of cholesterol in various types of samples.

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References

Xu J, Jiang D, Qin Y, Xia J, Jiang D, Chen H . CN Nanosheet Modified Microwell Array with Enhanced Electrochemiluminescence for Total Analysis of Cholesterol at Single Cells. Anal Chem. 2017; 89(4):2216-2220. DOI: 10.1021/acs.analchem.6b04635. View

Sekretaryova A, Beni V, Eriksson M, Karyakin A, Turner A, Vagin M . Cholesterol self-powered biosensor. Anal Chem. 2014; 86(19):9540-7. DOI: 10.1021/ac501699p. View

Chang H, Ho J . Gold Nanocluster-Assisted Fluorescent Detection for Hydrogen Peroxide and Cholesterol Based on the Inner Filter Effect of Gold Nanoparticles. Anal Chem. 2015; 87(20):10362-7. DOI: 10.1021/acs.analchem.5b02452. View

Nagaoka T, Tokonami S, Shiigi H, Matsumoto H, Takagi Y, Takahashi Y . Development of an electrochemical cholesterol sensor system for food analysis. Anal Sci. 2012; 28(2):187-91. DOI: 10.2116/analsci.28.187. View

Zhao C, Wan L, Jiang L, Wang Q, Jiao K . Highly sensitive and selective cholesterol biosensor based on direct electron transfer of hemoglobin. Anal Biochem. 2008; 383(1):25-30. DOI: 10.1016/j.ab.2008.08.022. View

Khatun R, Hunter H, Magcalas W, Sheng Y, Carpick B, Kirkitadze M . Nuclear Magnetic Resonance (NMR) Study for the Detection and Quantitation of Cholesterol in HSV529 Therapeutic Vaccine Candidate. Comput Struct Biotechnol J. 2017; 15:14-20. PMC: 5484764. DOI: 10.1016/j.csbj.2016.10.007. View

Huang Y, Tan J, Cui L, Zhou Z, Zhou S, Zhang Z . Graphene and Au NPs co-mediated enzymatic silver deposition for the ultrasensitive electrochemical detection of cholesterol. Biosens Bioelectron. 2017; 102:560-567. DOI: 10.1016/j.bios.2017.11.037. View

Gahlaut A, Hooda V, Dhull V, Hooda V . Recent approaches to ameliorate selectivity and sensitivity of enzyme based cholesterol biosensors: a review. Artif Cells Nanomed Biotechnol. 2017; 46(3):472-481. DOI: 10.1080/21691401.2017.1337028. View

Haggerty L, Tran D . Cholesterol Point-of-Care Testing for Community Pharmacies: A Review of the Current Literature. J Pharm Pract. 2016; 30(4):451-458. DOI: 10.1177/0897190016645023. View

10.

Rambaldi D, Reschiglian P, Zattoni A, Johann C . Enzymatic determination of cholesterol and triglycerides in serum lipoprotein profiles by asymmetrical flow field-flow fractionation with on-line, dual detection. Anal Chim Acta. 2009; 654(1):64-70. DOI: 10.1016/j.aca.2009.06.016. View

11.

Wu L, Cheng C . Flow-injection enzymatic analysis for glycerol and triacylglycerol. Anal Biochem. 2005; 346(2):234-40. DOI: 10.1016/j.ab.2005.08.031. View

12.

Ho J, Hsu H, Huang M . Liposome-based microcapillary immunosensor for detection of Escherichia coli O157:H7. Anal Biochem. 2004; 330(2):342-9. DOI: 10.1016/j.ab.2004.03.038. View

13.

Ho J, Huang M . Application of a liposomal bioluminescent label in the development of a flow injection immunoanalytical system. Anal Chem. 2005; 77(11):3431-6. DOI: 10.1021/ac0484474. View

14.

Chang Y, Fu C, Chen Y, Jou A, Chen C, Chou C . Use of liposomal amplifiers in total internal reflection fluorescence fiber-optic biosensors for protein detection. Biosens Bioelectron. 2015; 77:1201-7. DOI: 10.1016/j.bios.2015.10.012. View

15.

Dencher N, Heyn M . Formation and properties of bacteriorhodopsin monomers in the non-ionic detergents octyl-beta-D-glucoside and Triton X-100. FEBS Lett. 1978; 96(2):322-6. DOI: 10.1016/0014-5793(78)80427-x. View

16.

Wisitsoraat A, Sritongkham P, Karuwan C, Phokharatkul D, Maturos T, Tuantranont A . Fast cholesterol detection using flow injection microfluidic device with functionalized carbon nanotubes based electrochemical sensor. Biosens Bioelectron. 2010; 26(4):1514-20. DOI: 10.1016/j.bios.2010.07.101. View

17.

Mokwebo K, Oluwafemi O, Arotiba O . An Electrochemical Cholesterol Biosensor Based on A CdTe/CdSe/ZnSe Quantum Dots-Poly (Propylene Imine) Dendrimer Nanocomposite Immobilisation Layer. Sensors (Basel). 2018; 18(10). PMC: 6209991. DOI: 10.3390/s18103368. View

18.

Phetsang S, Jakmunee J, Mungkornasawakul P, Laocharoensuk R, Ounnunkad K . Sensitive amperometric biosensors for detection of glucose and cholesterol using a platinum/reduced graphene oxide/poly(3-aminobenzoic acid) film-modified screen-printed carbon electrode. Bioelectrochemistry. 2019; 127:125-135. DOI: 10.1016/j.bioelechem.2019.01.008. View