Bilirubin Direct 4+1 Clinical Chemistry reagent; Elitech

Description

INTENDED USE
ELITech Clinical Systems BILIRUBIN DIRECT 4+1 is intended for the quantitative in vitro diagnostic determination of direct bilirubin in human serum and plasma.

CLINICAL SIGNIFICANCE
Approximately 80-85 % of the bilirubin produced isnderived from the hememoiety of the haemoglobin released from aging erythrocytes in the reticuloendothelial cells. Bilirubin, bound to albumin, is transported into the liver where it is rapidly conjugated with glucuronide to increase its solubility. Then it is excreted into biliary canaliculi, and hydrolyzed in the gastrointestinal tract.
Unconjugated bilirubin serum concentration increases in case of overproduction of bilirubin (acute or chronic hemolytic anaemia) and in case of disorders of bilirubin metabolism and transport defects (impaired uptake by liver cells: Gilbert’s syndrome; defects in the conjugation reaction: Crigler-Najjar syndrome).
Reduced excretion (hepatocellular damage: hepatitis, cirrhosis…; Dubin-Jonhson and Rotor syndrome) and obstruction to the flow of bile (most often produced by gallstones or by tumors) induce an important elevation of conjugated bilirubin and in a minor extent an increase of unconjugated bilirubin (conjugated hyperbilirubinemia).

METHOD
Malloy-Evelyn modified. End point.

PRINCIPLE
Sulphanilic acid reacts with sodium nitrite to form diazotized sulphanilic acid. In the presence of accelerator (cetrimide), conjugated and unconjugated bilirubin react with diazotized sulphanilic acid to form azobilirubin (Bilirubin Total 4+1). In the absence of accelerator, only conjugated bilirubin reacts (Bilirubin Direct 4+1).
The increase of absorbance at 546 nm is proportional to bilirubin concentration.

REAGENTS COMPOSITION

Reagent 1: R1
Sulphanilic acid 29 mmol/L
BILIRUBIN TOTAL & DIRECT 4+1
Reagent 2: R2
Sodium nitrite 11 mmol/L

MATERIALS REQUIRED BUT NOT PROVIDED
– CALI-0550 ELICAL 2 4 x 3 mL
– CONT-0060 ELITROL I 10 x 5 mL
– CONT-0160 ELITROL II 10 x 5 mL
– Normal saline solution (NaCl 9 g/L)
– General Laboratory equipment.

– Do not use materials that are not required as indicated above.

WARNINGS AND PRECAUTIONS
– These reagents are for professional in vitro diagnostic use only.
– The reagents R1 contain sulphanilic acid . They may produce an allergic reaction.
– For more information, refer to the Safety Data Sheet (SDS).

– Take normal precautions and adhere to good laboratory practice.

– Use clean or single use laboratory equipment only to avoid contamination.
– Do not interchange reagent vials from different kits.

STABILITY OF REAGENTS
Store at 2-8 °C and protect from light. Do not freeze.
Do not use after expiration dates indicated on the vial labels.

On board stability:
The on-board stability is specific for each analyzer.
(Refer to § PERFORMANCE DATA).

PREPARATION
The reagents are ready to use.

REAGENT DETERIORATION
– The reagent R1 of Bilirubin Total 4+1 can be slightly cloudy. It contains a detergent that can lead to the formation of foam in washing units of some equipments. These two characteristics are without consequences on the product performances.
– The reagent R1 of Bilirubin Direct 4+1 and the reagent R2 of Bilirubin Total & Direct 4+1 should be clear.
Cloudiness would indicate deterioration.
– Do not use the product if there is visible evidence of biological, chemical or physical deterioration.

DAMAGED PACKAGING
Do not use the reagent if the damages of packaging might have an effect on the product performance (leakages, pierced vial).

SAMPLES
Specimen
– Serum or lithium heparinized plasma.
– Do not use other specimens.
Warnings and precautions
– For Direct Bilirubin : Do not use hemolysed samples.

– According to Good Laboratory Practice, venipuncture should be performed prior to the administration of drugs.
– Care must be taken to fill heparinized tubes according to the manufacturer’s instructions. An insufficient filling may lead to erroneous results.
– Protect the samples from light before and during the analysis.

Storage and stability
If plasma and serum are protected from light, samples are stable 1 day (Total Bilirubin) or 2 days
(Direct Bilirubin) at room temperature, 7 days at 2-8°C and 6 months at -20 °C.