MIDTERM LAB: LIPID PROFILE
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Requires at least _____ of fasting
10-16 hours
OPTIMAL FASTING:
12 HOURS
samples are not accepted
Hemolyzed and icteric
Specimen:
Serum or plasma EDTA
Not affected by fasting:
CHOLESTEROL determination AND HDL
Lipid Profile TEST:
Triglyceride determination Total Cholesterol determination HDL determination LDL determination (computed) VLDL determination (computed)
Friedewald Equation
TC = HDL + LDL + VLDL LDL = TC – HDL – TAG/5 (mg/dL) LDL = TC – HDL – TAG/2.175 (mmol/L)
Friedewald Equation FOR mg/dl:
LDL = TC – HDL – TAG/5 (mg/dL
Friedewald Equation FOR mmol/l:
LDL = TC – HDL – TAG/2.175 (mmol/L)
De Long Equation:
TC = HDL + LDL + VLDL LDL = TC – HDL – TAG/ 6.5 (mmol/L) LDL = TC – HDL – TAG/2.825 (mmol/L)
De Long Equation FOR mg/dl:
LDL = TC – HDL – TAG/ 6.5 (mmol/L)
De Long Equation FOR mmol/L:
LDL = TC – HDL – TAG/2.825 (mmol/L)
Cholesterol Measurement:
• Abell-Kendall Method • Abell, Levey, Brody Method • Sperry Method • Salkowski Method • Enzymatic Method • Isotope Dilution/Mass Spectrometry (IDMS)
OLD reference method
Abell-Kendall Method
3-step method
Abell-Kendall Method
Abell-Kendall Method reagent:
Libermann-Burchard reagent: gHAc, acetic anhydride, concentrated H2SO4
Abell-Kendall Method 3-step method:
Saponification -> Extraction -> Colorimetry
Current Reference Method
Abell, Levey, Brody Method
Abell-Kendall Method End Color:
GREEN
Abell-Kendall Method End Product:
cholestadienyl monosulfuric acid
Abell, Levey, Brody Method Reagents:
Libermann-Burchard rgt + KOH and alcoholic hexane
Abell, Levey, Brody Method End Color:
GREEN
Sperry Method 4-step method:
Saponification -> Extraction -> Colorimetry -> Purification with digitonide
4-step method
Sperry Method