Lab-on-a-chip for analysis of triglycerides based on a replaceable enzyme carrier using magnetic beads.
In this paper an enzyme-carrier-based microfluidic chip coupled with a gold nanoband microelectrode as electrochemical detector for Triglyceride (TG) determination was developed by co-immobilized lipase, Glycerokinase (GK) and glycerol-3-phosphate oxidase (GPOx) on chitosan/Fe(3)O(4) composite nanoparticles with a shell-core structure, which combined the advantageous features of microfluidic chips technology with magnetic beads. This procedure enabled the easy renewal of the microchip enzyme carrier after each determination in a highly reproducible manner. Several operational parameters such as working potential, buffer pH, adenosine triphosphate concentrations (ATP, mM), separation voltage and temperature were evaluated and optimized. The performance of enzyme-carrier-based microfluidic chip for TG determination was modulated by changing the length of enzyme carrier from 1.0 to 3.0 cm, and the linear ranges were changed from 0-4.0 mM to 0-10.0 mM with the detection limits from 15 μM to 6.0 μM. The enzyme carrier remained its 70% activity after 40 days storage. This system was successfully employed for on-line detection of TG in serums. The experimental results demonstrated that this enzyme carrier using magnetic beads based microfluidic chip provided a relatively simple, sensitive, miniature, and replaceable means for the accurate determination of TG in serum.
Chen
Shao-Peng
SP
Key Laboratory of Analytical Chemistry for Life Sciences, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210093, China.
Yu
Xiao-Dong
XD
Xu
Jing-Juan
JJ
Chen
Hong-Yuan
HY
D048271:Chitosan / Q000737:chemistry; D004563:Electrochemistry; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism; D052203:Ferrosoferric Oxide / Q000737:chemistry; D005991:Glycerol Kinase / Q000737:chemistry* / Q000378:metabolism; D005993:Glycerolphosphate Dehydrogenase / Q000737:chemistry* / Q000378:metabolism; D006046:Gold / Q000737:chemistry; D056656:Lab-On-A-Chip Devices; D008049:Lipase / Q000737:chemistry* / Q000378:metabolism; D008280:Magnetics; D008839:Microelectrodes; D053758:Nanoparticles / Q000737:chemistry; D014280:Triglycerides / Q000032:analysis*
D004800:Enzymes, Immobilized; D014280:Triglycerides; D006046:Gold; D048271:Chitosan; D005993:Glycerolphosphate Dehydrogenase; C098831:glycerol-3-phosphate oxidase; D005991:Glycerol Kinase; D008049:Lipase; D052203:Ferrosoferric Oxide