Note: – It is essential to use absolutely dry glassware for this estimation.
Requirements: – Std cholesterol soln (1 mg/ml). Chloroform, acetic anhydride, conc. H2SO4, blood or serum.
Procedure: – Prepare series of test tube, as given Add all the soln from burette mix the soln in each tube and plug with cork and keep in dark for 15 mins measure the co lour developed at 680 nm against blank.
Isolation of blood (or serum) cholesterol. Add 0.2 ml of serum or bld. to 10 ml. mix of acetone alcohol mix(1:1) in a centrifuge tube keep the tube in boiling water bath with intermittent shaking till the solvent begin to boil. Remove the tube the W.B.shake it for 10 min and centrifuge decant the supernatant liquid into a dry t.t.and evap. To dryness on a W.B. cool and dissolve in 2 ml chloroform. Use 1 ml for co lour development. Cholesterol level in blood is det to follow the treatment of diseases like hypo hyperthyroidism diabetes, coronary artery diseases and arthero scleroses.
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