This package insert follows the Pharmaceuticals, Medical devices and Other Therapeutic Products Act of Japan.
In Vitro Diagnostics
Marketing Approval No. 21200AMZ00383000** Revised: January 2017 (7th edition)* Revised: April 2008 (6th edition)

This package insert must be read carefully prior to use.
Bile acid assay kit(Classification No.: 30156000)

Assay Principle
1. Assay Principle
Bile acid in a sample is specifically oxidized by
3α-hydroxysteroid dehydrogenase (3α-HSD) andoxidized β-thionicotinamide adenine dinucleotide(Thio-NAD) to produce 3-ketosteroid and reduced
β-thionicotinamideadenine
dinucleotide(Thio-NADH). In the presence of 3α-HSD andreduced β-nicotinamide adenine dinucleotide(NADH), reaction with 3-ketosteroid produces
bile acid and oxidized β-nicotinamide adenine
dinucleotide (NAD). As mentioned above, tracelevels of bile acid are amplified by coenzyme
cycling, and the change of absorbance resulting
from Thio-NADH production is measured todetermine the concentration of bile acid.

Pureauto S TBA
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 be
guaranteed 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 ifrequired.
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.

Thio-NADBile acid
(3α-Hydroxysteroid)3-HSDNAD＊＊

Thio-NADH

3-Ketosteroid

NADH

2. Features
1) Liquid reagents, ready-to-use.
2) A highly sensitive enzyme cycling method isused.
3) The calibration curve is linear up to 180µmol/L.
4) Applicable to various automated analyzers.

Description (Kit Components)
Component: IngredientsTBA Substrate Solution 1:
Oxdized β-thionicotinamide
adenine dinucleotide(Thio-NAD)Good’s buffer
TBA Enzyme Solution 2:
Reduced β-Nicotinamide
adenine dinucleotide (NADH)
3α-Hydroxysteroid
dehydrogenase (3α-HSD)
Good’s buffer＊＊

Procedural Precautions ＊1. Properties of Samples and Sampling Methods
1) Samples
Serum may be used.
Serum bile acid levels are known to increase
after meals. Pay attention to the timing ofblood collection. If patients are not taking bileacid, collect blood early in the morning afterfasting.
2) Storage of samples4)
The isolated serum should be tested on thesame day.
Store samples at -20°C or lower.
Bring samples to room temperature (15–30C) before use.
2. Interfering substances
Assay results are not affected by free bilirubin (upto 50 mg/dL), conjugated bilirubin (up to 50mg/dL), hemoglobin (up to 500 mg/dL), ascorbic
acid (up to 100 mg/dL), or formazin turbidity (upto 3000 FTU).
3. Others
1) Always use bile acid standard solution forcalibration.

Intended Use
Measurement of bile acid in serum
Bile acid is produced from cholesterol in the liver.
Bile acid is mostly secreted into the bile as glycine ortaurine conjugates and are stored in the gallbladder.
Bile acid stored in the gallbladder is released into theintestine in response to stimulation such as foodintake, and promote the digestion and absorption offats, etc. Then most of the bile acid is reabsorbed in
the terminal ileum, transported to the liver via theportal vein, and taken up by hepatocytes(enterohepatic circulation).
Elevation of the concentration of bile acid in theperipheral blood specifically occurs in hepatobiliary
diseases. Bile acid levels closely reflect theexacerbation and recovery of these diseases.

1＊＊2) Precautions for assay range
If the concentration of sample exceeds assayrange, dilute the sample with saline and repeat
the measurement.
Dosage/Administration (Assay Procedure) ＊＊1. Preparation of reagents
Reagent (1): TBA Substrate Solution 1 is readyto use.
Reagent (2): TBA Enzyme Solution 2 is readyto use.
※After opening, TBA substrate solution 1 andTBA enzyme solution 2 can be used for 1 month
if kept in closed containers and stored at 2–10°C.
2. Assay Procedure
This product is compatible with Hitachi 9000
series automated analyzers. An example of theassay procedure is indicated below.

Sample+2.2 L37°C
Reagent 37C Reagent
Measurement(1)(2)
48–186 (Absorbance※)
150 L 5 min 50 μL
sec
Calculation ofconcentration※

Absorbance: The difference in absorbance
between 660 nm and 415 nm
Calibration material: Standard solution of bile
acid (Manufacture's assigned value)
Reagent blank: Purified water or saline

Assessment of Assay Results
1. Reference standard range≤ 10 μmol/L3)
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.
Performance
1. Sensitivity
1) Reagent blank: The change of absorbance is ≤0.020/min.
2) Sensitivity: The change of absorbance is0.020–0.100/ min per 50 µ mol/L of bile acid.
2. Accuracy: 90–110% of the expected assay value
3. Within-run Reproducibility:
Coefficient of variation ≤ 5%
(Test methods used for 1.–3. are in-housemethods.)
4. Measurement Range5): (On Hitachi 7170
automated analyzer)
2–180 μ mol/L
5. Correlatio5)
Serum N=69 r=0.999 y=1.01x+0.19
Control method: Approved in vitro diagnostic(enzyme cycling method)
6. Standard Material
Sodium glycochenodeoxycholate (in-housereference standard)

Precautions for Use or Handling ＊. Precautions for Handling (to Ensure Safety)
1) All samples used in the test should be handled
as a material possibly infected with HIV, HBV,HCV, or other viruses. To prevent infection, use
disposable gloves and avoid mouth pipetting
during the test.
2) The components of this product contain sodium
azide as a preservative. If any reagent is
accidentally ingested or comes into contact withthe eyes or skin, immediately rinse the area withwater and seek medical treatment, if necessary.
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 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 as
sodium hypochlorite solution at a concentrationof greater than 0.1%.
3) The reagents and treated samples should bediscarded 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 act
or related regulations.
5) The components of this product contains sodium
azide as a preservative. Sodium azide may reactwith lead or copper pipes and produce highly
explosive metallic azide. When disposing of thisproduct, flush it away with copious amounts ofwater.
4. Other precautions
1) Do not use the containers for other purposes.
2) Do not take apart the reagent cartridge before
use.
Storage and Shelf Life ＊＊1. Storage temperature: 2–10C
2. Shelf life: 1 year from the date of manufacture(The expiration date is printed on the outerpackage.)
Packaging ＊＊Name
TBA Substrate
Pureauto S TBA
Solution 1:
L set (Set for1 × 48 mL
HitachiTBA Enzyme
LABOSPECT
Solution 2:series)
1 × 16 mL

2Package

2＊＊References ＊1) Tanaka N. et al.: Jpn J Clin Med, 598 (extraedition), 521–524, 1989.
2) Fujiwara K.: Jpn J Clin Pathol, 37, 10, 1114–1121,1989.
3) Kanai M. (supervising editor): Kanai’s manual ofclinical laboratory medicine. 34th ed. 526,
Kanehara Shuppan, 2015.
4) Medical Practice Editorial Board: Laboratory testguide 2003–2004, 296, Bunkodo, 2013.
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, Japan3