HPLC Method for Separation of Bilirubin and Biliverdin on Newcrom R1 Column

Separation type: Liquid Chromatography Mixed-mode

Bilirubin and biliverdin are both pigments that are part of the heme degradation pathway in the body. They are structurally related but have different properties and functions.

Bilirubin : This is a yellow compound that is produced by the liver from the breakdown of old red blood cells. It is responsible for the yellow color of bruises and the yellow discoloration seen in conditions such as jaundice and liver disease. Bilirubin is transported in the blood to the liver, where it is conjugated with glucuronic acid and excreted in bile.

Biliverdin : This is a greenish pigment that is an intermediate product in the pathway that produces bilirubin. When red blood cells reach the end of their lifespan (about 120 days), they are broken down in the spleen. The heme group from the hemoglobin in these cells is converted into biliverdin by the enzyme heme oxygenase. This biliverdin is then quickly reduced to bilirubin by biliverdin reductase.

While both these compounds are normal products of metabolism, elevated levels can indicate disease. High levels of bilirubin, for example, can cause jaundice, a condition that results in yellowing of the skin and eyes. Furthermore, since these molecules are breakdown products of heme, they are part of the body’s natural antioxidant defense system. Bilirubin and biliverdin can help protect cells from damage by neutralizing harmful free radicals.

Bilirubin and biliverdin are both part of a class of compounds called tetrapyrroles. More specifically, they belong to a group of tetrapyrroles known as bile pigments.