Bilirubin liquicolor Calculation of the Bilirubin Concentration

If necessary, consider the reagent blank (see Notes).

Photometric Colorimetric Test for Total Normal assay Paediatric assay
Bilirubin Bilirubin conc. c [mg/dl] c [µmol/l] c [mg/dl] c [µmol/l]
DCA Method 546 nm, ∆Asample ∆Asample ∆Asample ∆Asample
Hg 546 nm x 12.5 x 214 x 58 x 992
Package Size
[REF] 10012 200 ml Complete Test Kit
Performance Characteristics
[IVD] Linearity
Method12 The linearity limit is for the normal assay 30 mg/dl or 510 µmol/l. At
concentrations of more than 20 mg/dl or 342 µmol/l the pediatric assay is
Albumin-bound bilirubin is released by a detergent. The total bilirubin
recommended to avoid too high absorbances.
reacts with diazotised 2,4-dichloroaniline to form a red azo dye as
indicator. Typical performance data can be found in the Verification Report,
accessible via
Contents www.human.de/data/gb/vr/su-bildc.pdf or
[DCA] 1 x 100 ml DCA Reagent www.human-de.com/data/gb/vr/su-bildc.pdf
2,4-Dichloroaniline 3.0 mmol/l
Hydrochloric acid 95 mmol/l Normal Values3
Detergent 70 g/l Adults: up to 1.1 mg/dl or 19 µmol/l
[NIT] 1 x 100 ml Nitrite reagent Newborn: up to 13.3 mg/dl or 227 µmol/l
Sodium nitrite 2.5 mmol/l
[BLK] 2 x 100 ml Sample blank reagent Quality Control
2,4 Dichloroaniline 1.5 mmol/l All control sera with bilirubin values determined by this method may be
Hydrochloric acid 47.5 mmol/l employed.
Detergent 35 g/l We recommend to use our quality control sera HumaTrol based on animal
serum or our SERODOS based on human serum.
Reagent Preparation
Mix [DCA] with [NIT] in a ratio of 1 + 1. Before use allow to stand for at Automation
least 15 min. at room temperature protected from light. Proposals to apply the reagents on analysers are available on request.
[BLK] is ready for use. Each laboratory has to validate the application in its own responsibility.

Reagent Stability Notes

The reagents are stable up to the stated expiry date when sealed and 1. The bilirubin readings may be too low if the sample is exposed to light,
stored at 2...25°C. and in hemolytic samples, due to the inhibition effect of hemoglobin on
the diazo reaction.
The working reagent is stable for 10 days at 15...25°C and for 21 days at
2...8°C when stored protected from light. 2. The working reagent may develop a weak colour during storage. This
will not affect the test performance. One reagent blank should, how-
Specimen ever, be run with each series. This is prepared by substituting specimen
Fresh, non-hemolytic serum, EDTA- or heparinised plasma. by water in the test procedure. If necessary, subtract the absorbance
Avoid hemolytic and lipemic samples! Samples must be protected from difference of the reagent blank (∆A reagent blank) from the absorbance
light. difference of each sample (∆Asample). If ∆A reagent blank is higher
than 0.040, fresh working reagent should be prepared.
Assay
References
Wavelength: 546 nm, Hg 546 nm
1. Rand R. N., di Pasqua A., Clin. Chem. 8, 570 (1962)
Optical path: 1 cm
2. Weigl E. et al., Med. Klin. 70, 664 (1975)
Temperature: 20...25°C
3. Schellong G., Icterus neonatorum, Thieme Verlag, Stuttgart (1962) 82
Measurement: against sample blank

Pipetting Scheme SU-BILDC INF 101301 GB 11-2008-15 |

Pipette into cuvettes
Normal assay Paediatric assay
Sample [BLK] Sample [BLK]
Specimen 100 µl 100 µl 20 µl 20 µl
working reagent 1000 µl --- 1000 µl ---
[BLK] --- 1000 µl --- 1000 µl
Mix and allow to stand for at least 10 min. at 20...25°C protected from
light. Measure absorbance of sample against the sample blank within
60 min.: ∆Asample = Asample – Asample blank

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