Development of co-immobilised enzymes amperometric biosensor for the determination of triglycerides in coconut milk
2020
Chittibabu, S. | Auddy, I. | Shanmugasundaram, S. | Manoj, D. | Nimbkar, S.
Enzyme biosensor was developed to determine the triglycerides present in coconut milk. Lipase, glycerol-3-phosphate oxidase (GPO) and glycerol kinase (GK) were co-immobilised on the gelatine membrane, and coated on the surface of the working electrode. The standardisation of the biosensor was carried out at different standard triolein solutions. The effect of gelatine concentration, glutaraldehyde concentration, and pH of reaction buffer were optimised. It was found that 45 mg gelatine, 2.5% glutaraldehyde concentration, and pH 7.0 yielded the best results. The developed biosensor could detect the triglyceride concentration between 0.3 and 1.3 mM. Triglyceride content in coconut milk was determined using the developed amperometric biosensor method, and the results obtained were compared with the triglyceride analysis in coconut milk by ultra-high performance liquid chromatography (UHPLC) method. It was found that the correlation between the results of the two methods was 97%. Apart from these, thermal stability and sensitivity loss on storage of the working electrode was also studied. The working electrode lost 48% of its activity in 4 h when maintained at 40°C, and lost 57.7% of its activity after 30 d when stored at 4°C.
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