Construction of an amperometric triglyceride biosensor using PVA membrane bound enzymes.
OBJECTIVE
A method is described for construction of an amperometric triglyceride (TG) biosensor using PVA membrane bound enzymes.


METHODS
A mixture of commercial lipase, glycerol kinase, glycerol-3-phosphate oxidase and horseradish peroxidase was co-immobilized onto polyvinyl alcohol (PVA) membrane through glutaraldehyde coupling. The biosensor measures current when polarized at 0.4 V.


RESULTS
The sensor showed optimum response within 2 s at pH 7.0 and 25 degrees C. The current (mA) was in proportion to concentration of TG in the range 0.56-2.25 mM. The minimum detection limit of the method was 0.21 mM. The analytic recovery of added TG was 94.3%. Within batch and between batch coefficients of variations (CV) were <5.85% and <4.13%, respectively. A good correlation (r=0.99) was found. Among various serum substances tested, only cholesterol caused slight stimulation (20%).


CONCLUSIONS
An amperometric method was developed for determination of TG employing this enzyme electrode.
Pundir
C S
CS
Biochemistry Research Laboratory, Department of Biochemistry, M D University, Rohtak-124 001, India. pundircs@rediffmail.com
Sandeep Singh
Bharvi
B
Narang
Jagriti
J
D015374:Biosensing Techniques / Q000295:instrumentation*; D055664:Electrochemical Techniques / Q000295:instrumentation*; D004800:Enzymes, Immobilized / Q000378:metabolism* / Q000648:ultrastructure; D005260:Female; D005991:Glycerol Kinase / Q000378:metabolism; D005993:Glycerolphosphate Dehydrogenase / Q000378:metabolism; D006735:Horseradish Peroxidase / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D057230:Limit of Detection; D008049:Lipase / Q000378:metabolism; D008297:Male; D008567:Membranes, Artificial; D008855:Microscopy, Electron, Scanning; D011142:Polyvinyl Alcohol / Q000737:chemistry*; D012015:Reference Standards; D013379:Substrate Specificity; D013696:Temperature; D014280:Triglycerides / Q000032:analysis*
D004800:Enzymes, Immobilized; D008567:Membranes, Artificial; D014280:Triglycerides; D011142:Polyvinyl Alcohol; D005993:Glycerolphosphate Dehydrogenase; C098831:glycerol-3-phosphate oxidase; D006735:Horseradish Peroxidase; D005991:Glycerol Kinase; D008049:Lipase