(M) Lec 4: Lipids and Lipoproteins (Part 2) Flashcards
TAGs (TESTING)
What sample is used: Plasma or Serum?
Both
TAGs (TESTING)
How long is the fasting period for TAG analysis?
12-14 hours
TAGs (TESTING)
TOF: Postural changes increase TAGs
False (decrease)
TAGs (TESTING)
Chemical Methods:
1. A fluorimetric method that produces a pink colored reaction
2. A colorimetric method that produces a blue colored reaction
A. Van Handel and Zilversmith
B. Hantzsch Condensation
- B
- A
TAGs (TESTING)
What is the enzymatic method called that is also known as the Bucolo and David Method?
Glycerol Kinase Method
TAGs (TESTING)
What is the CDC Reference Method for TAG analysis?
MODIFIED VAN HANDEL and ZILVERSMITH Method
TAGs (TESTING)
Modified Van Handel and Zilversmith Reference Method:
- Alkaline Hydrolysis
- Solvent Extraction
- Adsorption
- Colorimetry
- End Product
A. Formaldehyde
B. Alcoholic KOH
C. Silisic Acid
D. Chromotropic Acid
E. Chloroform
- B
- E
- C
- D
- A
TAGs (TESTING)
Silisilic Acid Chromatography isolates what?
TAGs
TAGs (TESTING)
Reference Values:
1. >500 mg/dL
2. 150-199 mg/dL
3. < 150 mg/dL
4. 499 mg/dL
A. Normal
B. Borderline High
C. High TAG
D. Very High TAG
- D
- B
- A
- C
TAGs (TESTING)
What is the conversion factor?
0.011 mmol/L
TAGs (TESTING)
Increased or Decreased?
- Hyperlipoproteinemia Type I, IIb, III, IV, V
- Alcoholism
- Nephrotic syndrome
- Hypothyroidism
- Pancreatitis
Increased
TAGs (TESTING)
Increased or Decreased?
- Malabsorption syndrome
- Hyperthyroidism
- Malnutrition burns
- Brain infarction
Decreased
Lipoproteins (TESTING)
What sample is used: Plasma or Serum?
Both
Lipoproteins (TESTING)
What 2 anticoagulants can be used for lipoprotein fractions?
SST and EDTA
Lipoproteins (TESTING)
What 2 parameters can be measured using a NON-FASTING sample?
HDL and TC
Lipoproteins (TESTING)
This method:
- is the reference method to quantify lipoproteins
- isolates the protein and TAG content of lipoproteins
- separation is based on density and weight
- reagent used is: potassium bromide with a 1.063 density
Utracentrifugation
Lipoproteins (TESTING)
Ultracentrifugation results using a potassium bromide reagent is expressed as what unit?
Svegberg Unit
Lipoproteins (TESTING)
This method:
- has a supporting medium of agarose gel (alam na)
- has the ff. staining dyes: Oil Red O, Fat Red 7B, and Sudan Black B
- moves from the most anodal to the least anodal portion of the media
Electrophoresis
Lipoproteins (TESTING)
The electrophoretic pattern of lipoproteins moves from most anodal to least anodal, order the ff. analytes in that manner:
- LDL
- VLDL
- Chylomicrons
- HDL
HDL (most anodal), VLDL, LDL, and Chylomicrons (least anodal)
Lipoproteins (TESTING)
This method:
- is an apolipoprotein assay that uses an antibody complex
- lipoprotein (a) is measured by a immunoturbidimetric assay
Immunoassay or Immunonephelometry
Lipoproteins (TESTING)
In chromatographic methods, what 2 types of chromatography is done?
Gel Chromatography or Affinity Chromatography
Lipoproteins (TESTING)
This method has the ff. principle:
The nuclei (sample) of atoms with characteristic spins when immersed in a static magnetic field and second oscillating magnetic field produce resonance or chemical shift that is measured and recorded.
Nuclear Magnetic Resonance Spectroscopy (NMRS)
Note: It measures HDL and LDL
Lipoproteins (TESTING)
What samples can be used in Nuclear Magnetic Resonance Spectroscopy (NMRS)? (3 answers)
- Serum
- EDTA Plasma
- Heparinized Plasma
Lipoproteins (TESTING)
What is the main disadvantage of Nuclear Magnetic Resonance Spectroscopy (NMRS)?
Requires a large volume of sample
Lipoproteins (TESTING)
This test is for the detection of increased chylomicrons and VLDL; the patient plasma stands overnight undisturbed in a refrigerator (@ 4ºC)
Standing Plasma Test/ Ref. Test/ Overnight Standing Test
Lipoproteins (TESTING)
Standing Plasma Test:
If (+) for chylomicrons, what is found on the top of the sample?
Milky/creamy layer
Lipoproteins (TESTING)
Standing Plasma Test:
If (+) for VLDL, what can be said about the turbidity of the sample?
Uniform/Homogenous
Lipoproteins (TESTING)
Standing Plasma Test:
If (+) for both Chylomicrons and VLDL, what can be said about the appearance of the sample?
It has both a creamy layer on top and uniform turbidity
Lipoproteins (TESTING)
This test uses the ff. reagents.
- heparin sulfates
- dextran sulfates
- phosphotungstate
- (+) divalent cations: Ca, Mg, and Mn
Polyanion precipitation
Lipoproteins (TESTING)
What is the most widely used reagent in the Polyanion Precipitation method?
Heparin Sulfate + Mn
Note: Mn removes interferences
Lipoproteins (TESTING)
The ff. parameters are the bases for what event in Polyanion Precipitation?
- Reagent conc.
- pH
- Ionic strength
- Lipid/Protein ratio
Separation
Lipoproteins (HDL TESTING)
This method of measuring HDLs:
- precipitates all Apo-B (100 and 48) containing lipoproteins such as chylomicrons, VLDL, LDL, and IDL
- liberated/remaining HDL is measured using a Chole reagent (used for cholesterol as well)
Polyanion Precipitation followed by Measurement of Supernatant Cholesterol
Lipoproteins (HDL TESTING)
This method of measuring HDLs:
- gets no prior treatment and separation procedures
- uses 2 reagents which: (1) prevents HDL from reacting [block non-HDL] and (2) releases HDL before measuring it enzymatically, respectively
Homogenous Assays
Lipoproteins (HDL TESTING)
What is the CDC’s reference 3-Step Method for HDL Testing?
- Ultracentrifugation
- Heparin Manganese Precipitation
- Abell-Kendall Assay