title: An amperometric cholesterol biosensor based on epoxy resin membrane bound cholesterol oxidase.
abstract: OBJECTIVE
  The use of epoxy resin membrane as a support for immobilization of enzyme has resulted into improved sensitivity and stability of biosensors for uric acid, ascorbic acid and polyphenols. The present work was aimed to prepare an improved amperometric biosensor for determination of serum cholesterol required in the diagnostics and management of certain pathological conditions.
  
  
  METHODS
  Epoxy resin membrane with immobilized cholesterol oxidase was mounted on the cleaned platinum (Pt) electrode with a parafilm to construct a working electrode. This working electrode along with Ag/AgCl as reference and Ag wire as an auxiliary electrode were connected through a three terminal electrometer to construct a cholesterol biosensor.
  
  
  RESULTS
  The sensor showed optimum response within 25 sec at pH 7.0 and 45°C. The linear working range of biosensor was 1.0 to 8.0 mM cholesterol. K m and I max for cholesterol were 5.0 mM and 9.09 μA, respectively. The biosensor measured serum cholesterol. The minimum detection limit of the sensor was 1.0 mM. The mean analytical recoveries of added cholesterol in serum (2.84 and 4.13 mM) were 91.4 ± 2.8 and 92.3 ± 3.1 per cent (n=6), respectively. Within and between assay coefficient of variation (CV) were <2 and <4 per cent, respectively. Biosensor had a storage life of 6 months at 4 o C.
  
  
  CONCLUSIONS
  The use of epoxy resin membrane as a support for immobilization of cholesterol oxidase has resulted into an improved amperometric cholesterol biosensor. The present biosensor had an advantage over the existing biosensors as it worked at comparatively lower potential.
authors.affiliation: Department of Biochemistry, M.D. University, Rohtak, India. pundircs@rediffmail.com
authors.forename: C S
authors.initials: CS
authors.lastname: Pundir
authors.forename: Jagriti
authors.initials: J
authors.lastname: Narang
authors.forename: Nidhi
authors.initials: N
authors.lastname: Chauhan
authors.forename: Preety
authors.initials: P
authors.lastname: Sharma
authors.forename: Renu
authors.initials: R
authors.lastname: Sharma
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D002789:Cholesterol Oxidase / Q000097:blood* / Q000378:metabolism; D003935:Diagnostic Techniques, Cardiovascular; D004566:Electrodes; D004800:Enzymes, Immobilized / Q000378:metabolism; D004853:Epoxy Resins; D006801:Humans; D006937:Hypercholesterolemia / Q000175:diagnosis*; D010984:Platinum; D013570:Synaptic Membranes / Q000737:chemistry / Q000378:metabolism*
chemical_list: D004800:Enzymes, Immobilized; D004853:Epoxy Resins; D010984:Platinum; D002789:Cholesterol Oxidase