bilirubin (conjugated/unconjugated) in serum

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Reference interval

Principle

The Kodak Ektachem Clinical Chemistry Slide (BuBc) is a dry multilayered, analytical element coated on a clear polyester support.

A 10 uL drop of sample is deposited on the slide & evenly deposited on the slide & evenly distributed by the spreading layer. Aided by caffeine, sodium benzoate, & surfactant in the spreading layer, Bu dissociated from albumin & migrated with Bc, through the masking layer to the gel registration layer. Proteins (including the albumin-bound Delta Bilirubin & hemoglobin), as well as lipids & lipocromes, are retained in the spreading layer. The masking layer optically blocks potentially interfering compounds trapped in the spreading layer, thereby, preventing them from being measured. In the gel registration layer, Bu & Bc bind to a cationic mordant. Bilirubin mono- & di-glucuronides, when bound to the mordant, have identical spectra & are quantitated together. As Bc in the vicinity of 400 to 420 nm, Bu & Bc have similar molar absorbtivities; at 460 nm, Bu has a higher molar absorbtivity than Bc. Because of these unique spectral characteristics, the reflection densities at two wavelengths (400 & 460 nm) are used to determine the concentrations of Bu & Bc

The following reaction sequences are involved:
Bilirubin complexes ------------> Bilirubin (Bu  + Bc)
Bilirubin + mordant ------------> Bilirubin-mordant complex

Preparation: No special patient preparation is necessary

Clinical significance

Increases

Specimen

Sample preparation: Collect specimen by standard venipuncture technique. Heparin may be used as an anticoagulant for plasma specimens. Sodium citrate, iodoacetate, sodium fluoride/potassium oxalate & EDTA anticoagulants should not be used.

Promptly remove serum or plasma from the clot or cells & analyze as soon as possible to minimize changes. Conversion of Bc to Bu has been observed, presumably due to hydrolysis of the glucuronide moieties. PROTECT SAMPLE FROM EXPOSURE TO LIGHT.

Handle specimens in stoppered containers to avoid contamination and evaporation. Refrigerate at 2-8 C if analysis is not performed within 4 hours after collection. Freeze specimens at -18 degrees C if analysis is delayed beyond 48 hours.

Minimum sample size of 0.5 milliliter: with an optimum size of 1.0 milliliter or larger.

More general terms

More specific terms

Additional terms

References

  1. Kodak Ektachem 700 Analyzer Operator's Manual, Kodak Clinical Products, Rochester, New York.
  2. Kodak Ektachem Slide Package Inserts, Kodak Clinical Products Rochester, New York.
  3. Kodak Ektachem Training Manual, Kodak Clinical Products, Rochester, New York.