Zhenhua GONG
Shanghai Jiaotong University, China
Title: Quantification of bilirubin on dry blood spot by tandem mass spectrometry
Biography
Biography: Zhenhua GONG
Abstract
BACKGROUND: Hyperbilirubinemia is caused by many kind diseases and is harmful to neonate development, it is necessary to detect hyperbilirubinemia as early as possible, or probably detect biliary atresia (BA) earlier. A new method for screening and clinical determination of blood bilirubin on a dry blood spot (DBS) using tandem mass spectrometry (MS/MS) was developed. METHODS: The serum bilirubin controls and blood were prepared as DBS, extracted into a methanol solution containing isotope-labeled external standards, were subjected to HPLC, detect by MS/MS. Multiple-reaction monitoring of m/z 585—299 for unconjugated bilirubin(UBIL), m/z761—585 for monoglucuronide bilirubin(BMG) and m/z 937—585 for diglucuronide bilirubin (BDG) were used to detect bilirubins. RESULTS: The recoveries of UBIL by MS/MS were 90%-120.31% with an R2 value of 0.928 after linear regression (p<0.001). The coefficients of variation were less than 20%.The blood UBIL in neonates aged 3-7 days (68.14±21.56umol/L) was higher than in children aged older than 1 mouth(38.24±7.95umol/L). BMG and BDG were not higher in BA than control significantly. CONCLUSION: Quantification of UBIL on a dry blood spot by MS/MS is accurate, reliable and feasible for screening and clinical tests. There may be δ-bilirubin caused direct bilirubin elevated in BA.