Bilirubin is an endogenous antioxidant in the human blood. While measurement of a total serum bilirubin, comprising of protein-bound (indirect bilirubin) and bilirubin glucuronide (direct bilirubin), is a routine clinical test, direct determination of free bilirubin represents an analytical challenge. Free bilirubin, which is the bioactive form, accounts for a minor fraction (<100 nM). Here we show the direct analysis of free bilirubin in human and animal blood serum samples using a state-of-the-art system comprised of newly developed high-performance liquid chromatography (HPLC) on reversed-phase (RP) C18 support coupled with thermal lens spectrometric detection (TLS), based on excitation wavelength of 457.9 nm using argon laser. The method excels in ultra-high sensitivity with limit of detection (LOD) and limit of quantitation (LOQ) of 90 pM and 250 pM, respectively. As a result, we present here the direct assay of free bilirubin in serum (around 10 nM), which can be employed as an assessment of antioxidative blood status.
Antioxidative blood status assessment by the free bilirubin assay
PASSAMONTI, SABINA;ZIBERNA, Lovro
2014
Abstract
Bilirubin is an endogenous antioxidant in the human blood. While measurement of a total serum bilirubin, comprising of protein-bound (indirect bilirubin) and bilirubin glucuronide (direct bilirubin), is a routine clinical test, direct determination of free bilirubin represents an analytical challenge. Free bilirubin, which is the bioactive form, accounts for a minor fraction (<100 nM). Here we show the direct analysis of free bilirubin in human and animal blood serum samples using a state-of-the-art system comprised of newly developed high-performance liquid chromatography (HPLC) on reversed-phase (RP) C18 support coupled with thermal lens spectrometric detection (TLS), based on excitation wavelength of 457.9 nm using argon laser. The method excels in ultra-high sensitivity with limit of detection (LOD) and limit of quantitation (LOQ) of 90 pM and 250 pM, respectively. As a result, we present here the direct assay of free bilirubin in serum (around 10 nM), which can be employed as an assessment of antioxidative blood status.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
