title: Development of cholesterol biosensor based on immobilized cholesterol esterase and cholesterol oxidase on oxygen electrode for the determination of total cholesterol in food samples.
abstract: The development of a cholesterol biosensor by co-immobilization of cholesterol esterase (ChEt) and cholesterol oxidase (ChOX) on oxygen electrode is described. The electrode consists of gold cathode and Ag/AgCl anode. The enzymes were immobilized by cross-linking with glutaraldehyde and Bovine Serum Albumin (BSA). The immobilized enzymatic membrane was attached to the tip of the electrode by a push cap system. The optimum pH and temperature of the sensor was determined, these are 6 and 25 degrees C respectively. The developed sensor was calibrated from 1-75 mg/dl of cholesterol palmiate and found linear in the range of 2-50 mg/dL. The calibration curve was drawn with V(i) (ppm/min)(initial velocity) vs different concentrations of cholesterol palmiate (mg/dL). The application of the sensor to determine the total cholesterol in different real food samples such as egg, meat was investigated. The immobilized enzymatic layer can be reused over 30 times and the stability of the enzymatic layer was studied up to 9 weeks.
authors.affiliation: Department of Food Technology and Biochemical Engineering, Jadavpur University, Kolkata 700 032, India.
authors.forename: Anjan Kumar
authors.initials: AK
authors.lastname: Basu
authors.forename: Parimal
authors.initials: P
authors.lastname: Chattopadhyay
authors.forename: Utpal
authors.initials: U
authors.lastname: Roychoudhuri
authors.forename: Runu
authors.initials: R
authors.lastname: Chakraborty
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D002784:Cholesterol / Q000032:analysis*; D002789:Cholesterol Oxidase / Q000737:chemistry*; D004563:Electrochemistry / Q000295:instrumentation* / Q000379:methods; D004566:Electrodes; D004800:Enzymes, Immobilized / Q000737:chemistry; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005504:Food Analysis / Q000295:instrumentation* / Q000379:methods; D005524:Food Technology / Q000295:instrumentation / Q000379:methods; D010100:Oxygen / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D002787:Sterol Esterase / Q000737:chemistry*
chemical_list: D004800:Enzymes, Immobilized; D002784:Cholesterol; D002789:Cholesterol Oxidase; D002787:Sterol Esterase; D010100:Oxygen