Which enzyme is NOT part of the enzymatic cholesterol method?

• A. Cholesterol Esterase
• B. Cholesterol Oxidase
• C. Peroxidase
• D. Lipase

Answer: (D)

In this enzymatic cholesterol method, the color signal comes from a specific cascade: cholesterol esters are first broken down to free cholesterol by cholesterol esterase, then cholesterol is oxidized by cholesterol oxidase to generate hydrogen peroxide, and finally peroxidase uses that hydrogen peroxide to oxidize a chromogen, producing the measurable color. Lipase isn’t part of this sequence because it acts on triglycerides, not cholesterol esters, so it wouldn’t create the necessary hydrogen peroxide or color signal.