Determination of serum triglycerides using lipase, glycerol kinase, glycerol-3-phosphate oxidase and peroxidase co-immobilized onto alkylamine glass beads

Abstract

A method for determination of serum triglycerides (Tgs) using lipase, glycerol kinase, glycerol-3-phosphate oxidase and peroxidase co-immobilized onto alkylamine glass beads (pore diameter 55 nm) through glutaralde¬hyde coupling was developed and evaluated. The minimum detection limit of the method was 0.54 mM. The analytical recovery of added triolein in the serum was 97.55±1.5% (mean ± S.D.). The mean value of serum Tgs, determined by the present method showed a good correlation (r=0.984) with the Bayer’s kit method, employing free enzymes. The within and between batch coefficients of variation (CV) were <2.25% and <1.35% respectively. No significant loss of activity was observed, when co-immobilized enzymes were reused for about 200 times and stored at 4°C in distilled water. The cost of Tg determination for 200 serum samples was less, as compared with Bayer’s kit method.