Cephalin-Cholesterol Flocculation Test

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WHAT IS CEPHALIN-CHOLESTEROL FLOCCULATION? Cephalin-cholesterol flocculation is a laboratory test for the non-specific measurement of blood globulins, which are a group of proteins that appear in abnormally high concentration (in which the abnormality is also known as hyperglobulinemia), in association with certain diseases. The test is carried out using HPLC (high-power liquid chromatography). Cephalin-cholesterol emulsion… Read More

Cephalin-Cholesterol Flocculation Test

WHAT IS CEPHALIN-CHOLESTEROL FLOCCULATION?

Cephalin-cholesterol flocculation is a laboratory test for the non-specific measurement of blood globulins, which are a group of proteins that appear in abnormally high concentration (in which the abnormality is also known as hyperglobulinemia), in association with certain diseases.

The test is carried out using HPLC (high-power liquid chromatography). Cephalin-cholesterol emulsion by pathological sera using HPLC can be carried out.

This test is similar to the thymol turbidity test and has become outdated due to the cumbersome and unclear nature of its procedure.

PROCEDURE

The separation of bilirubin species in serum by HPLC is confused by the presence of a high concentration of proteins (albumin, in particular). 

For the analysis of many substances in serum in HPLC, these proteins are always removed before chromatography. In the case of bilirubin in pathological sera, this results in the loss of variable, and most times, large fractions of the total amount of bilirubin present. 

To avoid this problem, a reversed-phase elution system is applied. Linear gradient elusion (1.4ml/min) at 41°C from a pH 2.0 phosphate buffer to one containing 80% isopropyl alcohol, both with 5% methoxy ethanol (v/v), enables the separation of unconjugated bilirubin sample in albumin solution on a reversed-phase HPLC column. 

Unfortunately, other proteins present in the serum samples do not elute from the column and cause its rapid deterioration. To stop this occurrence, a pre-chromatography step should be applied to serum samples to remove higher molecular weight proteins, specifically the globulins. 

First, the serum is diluted with a 10% human serum albumin solution (w/v), heated, and treated at 17°C with a 27.7% solution of Na₂SO₄ (w/v) to precipitate primarily higher molecule-weight proteins. 

The globulins are removed from the sample, while most of the albumins and the associated bilirubin species are retained in the aqueous solution.

HYPERGLOBULINEMIA

This is a condition characterized by abnormally large amounts of globulins in the blood. Among associated conditions are multiple myeloma, collagen disease, liver cirrhosis, sarcoidosis, kala-azar, lymphogranuloma, referendum, and certain other infections.

HYPERGAMMAGLOBULINEMIA

This is an uncommon condition that is usually the result of an infection. It is characterized by elevated levels of immunoglobulins in one’s blood. People with the condition mostly have increased levels of IgG. Other diseases associated with it include autoimmune disorder, malignancy (such as multiple myeloma), among others. 

CONCLUSION

Developments have led to many types of serological tests alongside flocculation tests. 

Other types of serum test include:

  • Neutralization tests
  • Hemagglutinin-inhibition tests
  • Enzyme-linked immunosorbent assays (ELISAs)
  • Chemiluminescence immunoassays.

Today, attention has been shifted to other scientifically simplified methods of serological determinations, this denying flocculation test more research information and perhaps clinical applications.

FAQs

  • What are the symptoms of high protein in the blood?

The indications of high levels of protein in your body may include pain in the bones, weight loss, as well as loss of appetite, excessive thirst (via frequent dehydration), frequent infections, and numbness or tingling (in your hands, legs, and feet).

  1. What is the normal range for globulin?

Normal value ranges are:

Serum globulin = 2.0gm/dL to 3.3gm/dL.

IgM = 75mg/dL to 300mg/dL.

  • What does it mean if your IgG is high?

High levels of immunoglobulin G (IgG) may seem to pose a long-term chronic infection such as HIV. Levels of IgG also get higher in IgG multiple myeloma, long-term hepatitis, and multiple sclerosis.

  • What is a high IgM reading?

Patients may have a slightly higher risk of gastrointestinal diseases (including celiac disease), autoimmune disease, or a modest increase in the rate of superficial infections. 

High IgM is ˃2g/L. elevated IgM levels are a non-specific marker of inflammation, but can be associated with liver disease. Low IgM is ˂0.5g/L.

  • Why does globulin increase in cirrhosis?

The serum protein changes occurring in the liver disease associated with parenchymal damage are decreased in serum albumin and increased in gamma globulin levels. Beta-gamma bridging in the electrophoretogram is highly characteristic of hepatic cirrhosis.