Determination of serum triglyceride by enzyme electrode using covalently immobilized enzyme on egg shell membrane

• Published in: International journal of biological macromolecules 2010-12, Vol.47 (5), p.691-695
• Main Authors: Narang, Jagriti, Minakshi, Bhambi, Manu, Pundir, C.S.
• Format: Article
• Language: English
• Subjects: Animals; Biosensing Techniques - methods; Biosensor lipase; Chickens; Egg Shell - metabolism; Egg Shell - ultrastructure; Egg shell membrane; Enzymes, Immobilized - metabolism; Female; Glycerol Kinase - metabolism; Glycerolphosphate Dehydrogenase - metabolism; GPO; Humans; Hydrogen-Ion Concentration; Kinetics; Limit of Detection; Lipase - metabolism; Male; Membranes; Reference Standards; Substrate Specificity; Temperature; Triglycerides - blood
• ISSN: 0141-8130; 1879-0003
• DOI: 10.1016/j.ijbiomac.2010.09.001

A mixture of commercial lipase, glycerol kinase and glycerol-3-phosphate oxidase was co-immobilized onto egg shell membrane through covalent coupling. A method is described for fabrication of a triglyceride (TG) biosensor using egg shell membrane bound enzymes. The biosensor measured current, i.e. flow of electrons generated from H 2O 2, maximally when polarized at 0.4 V. The biosensor showed optimum response within 10 sec at pH 7.0 and 35 °C. The current was in proportion to concentration of TG in the range 0.56–2.25 mM. An amperometric method was developed for determination of TG employing this enzyme electrode. The minimum detection limit of the method was 0.28 mM. The analytic recovery of added TG was 95.00% and 96.50%. Within batch and between batch coefficients of variations (CV) were