What is Bilirubin?

Much of medicine revolves around understanding complex biological substances. Among these varied substances, Bilirubin is of particular interest. Like many biological substances, this pigment finds its origin in the breakdown of a certain molecule, in this case, heme, a component of hemoglobin. Hemoglobin is responsible for transporting oxygen in the blood. When hemoglobin breaks down, about 80% becomes Bilirubin. Healthy bodies efficiently manage this normal process by excreting the Bilirubin through bile, a crucial digestive fluid produced by the liver. But if the system becomes inefficient, for either the production, transportation, or excretion of Bilirubin, it can lead to jaundice which is characterized by yellowing of skin and eyes.

Bilirubin is a linear tetrapyrrole and an organic derivative of hemoglobin degradation. It exists in two forms in the human body: unconjugated (or free) and conjugated (or direct). The former is insoluble in water and toxic, while the latter is a non-toxic form that is soluble in water. High levels of unconjugated Bilirubin may cause brain damage, primarily in newborn infants, a condition referred to as kernicterus. Hence, it is crucial to quickly and efficiently detect Bilirubin levels in the body.

Bilirubin detection methods and principles

Several clinical diagnostic methods are employed to detect and measure Bilirubin levels in blood, all of which are based on different scientific principles. Spectrophotometry is the most commonly used technique. This technique is non-invasive and takes advantage of Bilirubin's capability to absorb light at certain wavelengths. The light intensity changes are then measured and translated into Bilirubin concentration.

High-Performance Liquid Chromatography (HPLC) is another method used to detect Bilirubin. This technology involves passing a sample mixture through a column filled with a solid adsorbent material. Each component in the sample interacts differently with the adsorbent material, causing different flow rates for different components and leading to the separation of components as they exit the column.

Atomic Absorption Spectroscopy (AAS) can also be used in Bilirubin detection. Atomic absorption spectroscopy is a spectroanalytical procedure for the qualitative detection of chemical substances that use the absorption of light by free metallic ions. When a photon of a specific energy level encounters an atom of the same energy level, the atom absorbs the photon and elevates its energy level.

Laboratory diagnostic companies also utilize immunoassays for Bilirubin determination. This biochemical test measures the concentration of a substance in a biological liquid, such as serum or urine, using the reaction of an antibody or antibodies to its antigen.

Bilirubin Assay Kits:

The market is replete with several Bilirubin assay kits, each exploiting different scientific principles and techniques for the detection and measurement of Bilirubin.

Bilirubin Assay Kit (Colorimetric): This kit operates on the basis of the Jendrassik-Grof method for the quantification of total, direct and indirect Bilirubin.

Total Bilirubin Assay Kit: This kit is used for the quantitative analysis of total bilirubin concentration in neonatal serum.

Bilirubin (Total and Direct) Colorimetric Assay Kit: This assay kit aids quantification of Bilirubin in research environments.

Bilirubin Direct Assay Kit: Making use of colorimetric detection, this kit measures direct Bilirubin.

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