5. .

Direct Spectophotometry
This method measurement is based on the absorption of light by bilirubin near 460 nm and is restricted primarily to blood specimens from healthy newborns in which unconjugated bilirubin is the predominant species. 2. HPLC HPLC methods rapidly separate and quantify the four bilirubin fractions. It uses Micronex RP-30 column, which does not require salting out of globins or chemical transformation of the bilirubin conjugates. 3. Enzymatic methods Direct bilirubin is measured at ph 3.7 to 4.5 at thisph range, the enzyme oxidizes bilirubin conjugates . 4. Diazymes Direct Bilirubin Vanadate Oxidation assay is a dual stable liquid method that provides an enhance level of accuracy, reliability and freedom from interference compared to conventional Diazo procedures. The test offers an extended testing range of 0.1 to 40 mg/dL provides excellent precision (%CV <5%) and demonstrates no significant interference from ascorbate to 50 mg/dL and hemoglobin up to at least 500 mg/dL source: Tietz, fundamentals of clinical chemistry