Determination of Serum Cholesterol in Biochemistry
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Questions and Answers

What is the purpose of the enzyme used in the cholesterol test?

  • To catalyze the reaction between hydrogen peroxide and 4-aminoantipyrine
  • To convert cholesterol into cholestenone
  • To measure the concentration of cholesterol in the sample
  • To break down cholesterol esters into cholesterol and fatty acids (correct)
  • What is the wavelength used to measure the absorbance of the sample in the cholesterol test?

  • 450 nm
  • 500 nm (correct)
  • 600 nm
  • 550 nm
  • What is the purpose of the standard used in the cholesterol test?

  • To calibrate the spectrophotometer
  • To provide a known concentration of cholesterol for comparison
  • To ensure the accuracy of the test
  • All of the above (correct)
  • What is the recommended temperature for incubating the samples in the cholesterol test?

    <p>37ºC</p> Signup and view all the answers

    How long should the samples be incubated in the cholesterol test?

    <p>10 minutes</p> Signup and view all the answers

    What is the conversion factor used to convert cholesterol concentration from mg/dL to mmol/L?

    <p>0.0258</p> Signup and view all the answers

    What is the normal range for total cholesterol in adults?

    <p>Less than 200 mg/dL</p> Signup and view all the answers

    What is the recommended storage condition for cholesterol samples?

    <p>2-8ºC</p> Signup and view all the answers

    What is the primary role of HDL cholesterol in the body?

    <p>To carry cholesterol back to the liver</p> Signup and view all the answers

    What percentage of plasma cholesterol is esterified with fatty acids to form cholesterol esters?

    <p>Two-thirds</p> Signup and view all the answers

    Which type of cholesterol is referred to as 'bad cholesterol'?

    <p>LDL cholesterol</p> Signup and view all the answers

    What is the significance of determining serum cholesterol levels?

    <p>To estimate risk of cardiovascular disease</p> Signup and view all the answers

    Which of the following is NOT a risk factor for cardiovascular disease?

    <p>High physical activity levels</p> Signup and view all the answers

    Which of these is a source of cholesterol in the body?

    <p>Diet and liver production</p> Signup and view all the answers

    Study Notes

    Cholesterol

    • Cholesterol is essential for cell membranes, hormones, and bile acids formation
    • It is obtained from both diet and liver production
    • Cholesterol is carried in the blood attached to proteins, forming lipoproteins

    Types of Cholesterol

    • LDL cholesterol (bad cholesterol) is part of low-density lipoprotein
    • HDL cholesterol (good cholesterol) is part of high-density lipoprotein, helps prevent disease by carrying cholesterol away from cells to the liver for breakdown and removal

    Importance of Determining Serum Cholesterol

    • Estimating risk of cardiovascular disease
    • Diagnosis and classification of lipemia
    • Requested for individuals with risk factors such as:
      • Family history of high cholesterol or heart disease
      • Being overweight or obese
      • Diabetes mellitus
      • High blood pressure (hypertension)
      • Smoking cigarettes

    Principle of the Method

    • Enzymatic colorimetric method used to determine total serum cholesterol level
    • Involves cholesterol esters, cholesterol, and fatty acids reaction
    • Measured spectrophotometrically at λ=500 nm and compared to a standard of known cholesterol concentration (200 mg/dL)

    Test Preparation

    • No fasting required for a cholesterol test
    • Sometimes a fasting sample is required for a full lipid profile including cholesterol
    • Sample (serum or plasma) requirements:
      • Stability of the sample for 7 days at 2-8ºC
      • Freezing at –20ºC will keep samples stable for 3 months

    Procedure

    • Assay conditions:
      • Wavelength: 505 nm (500-550)
      • Cuvette: 1 cm light path
      • Temperature: 37ºC /15-25ºC
    • Reagents and sample preparation:
      • Blank, standard, and sample added to cuvette
      • Mix and incubate for 5 min at 37ºC or 10 min at room temperature
      • Read absorbance (A) of samples and standard against the blank

    Calculation

    • Conversion factor: mg/dL x 0.0258= mmol/L
    • Ranges for total cholesterol in adults:
      • Normal: Less than 200 mg/dL
      • Borderline high: 200 to 239 mg/dL

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    Description

    This quiz assesses knowledge on the determination of serum cholesterol, its importance in forming cell membranes and hormones, and its production in the liver.

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