This package insert follows the Pharmaceuticals, Medical devices and Other Therapeutic Products Act of Japan.
In Vitro Diagnostics
Marketing Approval No. 20300AMZ00140000** Revised: January 2017 (9th edition)* Revised: April 2008 (8th edition)

This package insert must be read carefully prior to use.
Class II General and Biochemical Test Series(Classification No.: 80022002)
Direct bilirubin assay kit(Classification No.: 30157002)

1) Because self-sample blank method is used,
analytical results are hardly affected byinterfering substance.
2) Instruments are not damaged.
Procedural Precautions ＊1. Properties of Samples and Sampling Methods
1) Samples
Serum may be used.
2) Storage of samples
If the isolated serum or plasma sample cannot
be tested on the same day, specimens should
be stored as follows:
2–10°C: for tests within 1 week≤- 20°C: for tests after more than 1 week
Bring samples to room temperature (15–30°C) before use.
Bilirubin is isomerized by light (both artificialand solar), resulting is discoloration. Store
under light shielding.
2. Interfering substances
Hemolyzed and chyle samples give positive errors
to analytical results. Perform remeasurement bydifferent methods, if the analytical result is
extremely higher than those of other related
variables.
3. Others
1) For calibration, use the FACTOR obtainedwith total bilirubin as the conversion factor.
2) Precautions for assay range
If the concentration of sample exceeds assay
range, dilute the sample with saline and repeat
the measurement.
3) Do not perform the assay under direct
sunlight.

Clinimate D-BIL-2
General Precautions
1. This product is for in vitro diagnostic use, andmust not be used for any other purposes.
2. Clinicians should make a comprehensive clinicaldecision based on assay results in conjunction withclinical symptoms and other examination results.
3. This product should be used only as directed in thispackage insert. Reliability of results cannot beguaranteed if there are any deviations from theinstructions in this package insert.
4. If the reagent accidentally comes in contact witheyes and/or mouth, rinse immediately with ample
water as first aid, and consult the doctor if
required.
5. Carefully read the operating instructions for eachtype of automated analyzers prior to using thisproduct. Parameters for each type of analyzers areavailable, and can be requested from SEKISUIMEDICAL CO., LTD. if required.
6. Perform a quality control test prior to assay toensure accuracy.
Description (Kit Components) ＊＊Component: Ingredients
D-BIL-2 Buffer Solution 1: Glycine buffer
BIL・D-BIL-2 Reactant Reagent 2: Sodium nitrite
BIL・D-BIL-2 Dissolution Solution 2:
Sulfanilic acid
Hydric acid

Dosage/Administration (Assay Procedure) ＊＊1. Preparation of reagents
Reagent (1): D-BIL-2 Buffer Solution 1 is readyto use.
Reagent (2): Dissolve BIL ・ D-BIL-2 Reactant
Reagent 2 with BIL ・ D-BIL-2 Dissolution
Solution 2.
Reagent (2) is stable for 7 days after preparation
when stored at 2–10°C with protection from light.
2. Assay Procedure
This product is compatible with various types ofautomated analyzers. An example of the assay
procedure is indicated below.

Intended Use
Measurement of direct bilirubin in serum
The total amount of serum bilirubin varies according
to the disintegration of hemoglobin and depending on
the condition of metabolism in the liver. On the otherhand, the direct to indirect bilirubin ratio plays animportant role in the diagnosis and course evaluationof hepatobiliary and hematological diseases.
Assay Principle
1. Assay Principle
Chemical method (azobilirubin method)
Direct bilirubin (conjugated bilirubin) in samples
reacts with diazo solution and formredazobilirubin. The direct bilirubin content isdetermined by measuring the absorbance of thisred sample.
Diazo solution
Direct bilirubin in sample→Azobilirubin (red)Sample Reagent (1) 37C Measurement+────▶
※10 L350 L
3 min (Absorbance I )37CReagent (2) ────▶ Measurement
4.5 min (Absorbance II※)
70 L────▶

2. Features

1

Calculation ofconcentration※

discarded as medical waste or industrial waste
according to the waste disposal regulations.
4) The reagents should be disposed of inaccordance with the Water Pollution Control actor related regulations.
4. Other precautions
Do not use the containers for other purposes.

Absorbance I and II: The difference in absorbance
between 600 nm and 546 nm.
Calibration method: Conversion factor (FACTORfor total bilirubin)
Reagent blank: Purified water or saline

Assessment of Assay Results
1. Reference standard range2)
0–0.6 mg/dL
2. There may be reactions or interfering reactions
with non-target substances. If assay resultsappear to be unreliable, repeat the measurement(if necessary, after dilution) or try anotheranalytical methods.

Storage and Shelf Life ＊＊Storage
Shelf life
temperature
D-BIL-2
1 year from
2–10C
Bufferthe date ofSolution 1
manufacture
2 years from
BIL・ D-BIL-2 2–10CClinimate
the date ofReactant
D-BIL-2
manufacture
Reagent 2Room2 years fromBIL・ D-BIL-2
temperature the date ofDissolution
manufacture
Solution 2
NamePerformance
1. Sensitivity
1) Reagent blank: absorbance being equal to or
lower than 0.02
2) Sensitivity: The absorbance is 0.19–0.26 per
10 mg/dL of bilirubin.
2. Accuracy: 90–110% of the expected assay value
3. Within-run Reproducibility:
Coefficient of variation ≤ 3%
(Test methods used for 1. –3. are in-housemethods.)
4. Measurement Range5): (On Hitachi 7170S
automated analyzer)
0.05–20.0 mg/dL
5. Correlatio5)
Serum N=55 r=0.999 y=0.92x–0.13
Control method: Approved in vitro diagnostics(Alkaline azobilirubin method)
6. Standard Material
SRM916a（NIST）＊When the above reagents are combined in use, referto the information about storage and expiration datelisted on the outer package.)
Packaging
NamePackage(1)

D-BIL-2
Buffer Solution 1

4 × 100 mL(2)

BIL・D-BIL-2
Reactant Reagent 2
BIL・D-BIL-2
Dissolution Solution 2

Clinimate
D-BIL-2

4 × for 50 mL
4 × 50 mL

Constituent reagents are available in otherconfigurations. For further details please contact
SEKISUI MEDICAL CO., LTD.

＊Precautions for Use or Handling ＊1. Precautions for Handling (to Ensure Safety)
All samples used in the test should be handled as amaterial possibly infected with HIV, HBV, HCV,or other viruses. To prevent infection, use
disposable gloves and avoid mouth pipetting
during the test.
2. Precautions for use
1) This product should be stored as directed,
without freezing. Freezing can deteriorate thereagents, which can produce inaccurate results.
Therefore, avoid using the reagents which havebeen previously frozen.
2) Do not use expired reagents. Use of suchreagents cannot guarantee the reliability ofmeasurement values.
3) Do not replenish the reagents.
4) Do not perform the assay under direct sunlight
3. Precautions for Disposal
1) Before disposal, used samples and theircontainers must be immersed in sodium
hypochlorite solution at a concentration ofgreater than 0.1% for longer than 1 hour or
autoclaved at 121ºC for 20 minutes.
2）To prevent infections from spilled samples orsolutions containing samples, wipe the spilledarea thoroughly with disinfectants such assodium hypochlorite solution at a concentrationof greater than 0.1%.
3) The reagents and treated samples should be

References ＊1) Michaelson, M.: Scand. J. Clin. & Lab. Invest, 13,Supple, 56, 1961.
2) Sasaki M. et al.: Sampling of constituents of thehuman body, 79, Kodansha, 1972.
3) Kitamura M. (author and editor): Practical
Clinical Chemistry, 277, Ishiyaku Shuppan,
1974.
4) Kanai I and Kanai M. (author and editor):
Kanai’s manual of clinical laboratory medicine.
32nd ed. 577, Kanehara Shuppan, 2005.
5) In house data, SEKISUI MEDICAL CO., LTD.
Contact ＊SEKISUI MEDICAL CO., LTD.
international@sekisui.com＊ ＊Manufacturer ＊SEKISUI MEDICAL CO., LTD.
1-3, Nihonbashi 2-chome, Chuo-ku, Tokyo, Japan2