This package insert follows the Pharmaceuticals, Medical devices and Other Therapeutic Products Act of Japan.
In Vitro Diagnostics
Marketing Notification No. 13A2X00197218045**Revised: January 2017 (6th edition)*Revised: September 2013 (5th edition)

This package insert must be read carefully prior to use.
Cholesterol assay kit(Classification No.: 30159000)

presence of peroxidase (POD) to form a
red-purple color. Total cholesterol content isdetermined by measuring the absorbance of thesample. The influence of ascorbic acid is blocked
by ascorbate oxidase.
CEEsterified cholesterol ─▶ Free cholesterol + Fatty
acid
CODFree cholesterol ─▶ Δ4-cholestenone + H2O2POD
H2O2 + ESBmT + 4-Aminoantipyrine ─▶
Red-purple color
2. Features
1) Assay results are hardly affected by crosscontamination.
2) Bilirubin, hemolysis, ascorbic acid and chylehave minimal effects on results.

QUALIGENT CHO
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 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.

Cholesterol in serum or plasma is derived fromdietary intake and is also synthesized in the liver.
Measurement of cholesterol is important as an index
of abnormal lipid metabolism and is also considered
to be useful for the diagnosis of hepatic dysfunction
and metabolic disease.

Procedural Precautions ＊1. Properties of Samples and Sampling Methods
1) Samples
Serum and plasma (heparin plasma, EDTA
plasma and citrated plasma) may be used.
2) Storage of samples1)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.
2. Interfering substances
Assay results are not affected by free bilirubin (upto 20 mg/dL), conjugated bilirubin (up to 20mg/dL), hemoglobin (up to 500 mg/dL), ascorbic
acid (up to 50 mg/dL), formazin turbidity (up to3000 FTU), or Intralipos (up to 5%).
3. Others
1) Always use Cholestest N Calibrator forcalibration.
However, for Hitachi LABOSPECT 008
Automated Analyzer, use QUALIGENT N
Calibrator (manufactured by SEKISUIMEDICAL) for calibration.
2) Precautions for assay range
If the concentration of sample exceeds assay
range, dilute the sample with saline and repeat
the measurement.

Assay Principle
1. Assay Principle
Cholesterol exists in samples as both the esterified
and free types. Esterified cholesterol is converted
to free cholesterol by the action of cholesterolesterase (CE). Then free cholesterol is oxidized bycholesterol oxidase (COD) to form hydrogenperoxide. Hydrogen peroxide causes oxidative
condensationof4-aminoantipyrine
andN-ethyl-N-sulfobutyl-m-toluidine (ESBmT) in the

Dosage/Administration (Assay Procedure) ＊1. Preparation of reagents
Reagent (1): CHO Enzyme Solution 1
is ready to use.
Reagent (2): CHO Enzyme Solution 2
is ready to use.
2. Assay Procedure
This product is compatible with Hitachi 9000
series and LABOSPECT series automatedanalyzers. Assay procedure is indicated below.

Description (Kit Components) ＊＊Component: Ingredients
CHO Enzyme Solution 1:4-Aminoantipyrine
Cholesterol esterase
Peroxidase
CHO Enzyme Solution 2:
Cholesterol oxidase (microbialorigin)
N-ethyl-N-sulfobutyl-m-toluidine
sodiumIntended Use
Measurement of total cholesterol in serum orplasma＊ ＊1Sample+2 L

Reagent(1)
150 L
Reagent(2) 50 μL

37C
────▶
5 min37C────▶
5 min────▶

※antibodies. However, these reagents (as well asthe samples) should be considered potentially
infectious and handled with great care.
3) Proclin 300, which possesses skin-irritative
potential, is added as an antiseptic agent in theCHO Enzyme Solution 1. Therefore, if thereagent comes in contact with skin or clothes,rinse immediately with ample water, and consult
the doctor if skin irritation develops.
4) Sodium azide is added as an antiseptic agent in
the CHO Enzyme Solution 2. Therefore, if thereagent comes in accidentally contact with eyes,mouth or skin, rinse immediately with ample
water as first aid, and consult the doctor ifrequired.
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) Sodium azide is added as an antiseptic agent in
the CHO Enzyme Solution 2. It can react withlead or copper pipes to produce the highly
explosive metal azide. Therefore, the reagent
should be flushed with large amounts of water
when disposing.
4. Other precautions
1) Do not use the containers for other purposes.
2) Do not take apart the reagent cartridge before.

Measurement(Absorbance I※）
Measurement(Absorbance II※)
Calculate ofconcentration

Absorbance I and II: The absorbance difference
between 800 nm and 600 nm.
Calibration material: Cholestest N Calibrator
Use QUALIGENT N Calibrator (manufactured bySEKISUI MEDICAL) for Hitachi LABOSPECT
008 Automated Analyzer
for calibration.
(Manufacture’s assigned values)
Reagent blank: Purified water or saline

Assessment of Assay Results ＊＊1. Reference standard range2)
142–248 mg/dL
2. Diagnostic criterion3)
Hypercholesterolemia: ≥ 220 mg/dL
3. There may be reactions or interfering reactions
with non-target substances. If assay results appear
to be unreliable, repeat the measurement (ifnecessary, after dilution) or try another analyticalmethods.
Performance ＊＊1. Sensitivity
1) Reagent blank: absorbance being equal to or
lower than 0.052) Sensitivity: The absorbance is 0.40–0.72 per300 mg/dL of cholesterol.
2. Accuracy: 95–105% of the expected assay value
3. Within-run Reproducibility:
Coefficient of variation ≤ 3%
(Test methods used for 1.-3. are in-housemethods.)
4. Measurement Range4): (On a Hitachi 9000 series
automated analyzer)
5–1000 mg/dL
5. Correlation 4)
1) Serum
N=70 r=0.997 y=1.01x–2.06
Control method: Approved in vitro diagnostic(enzymatic method)
2) Plasma
N=50 r=0.999 y=1.00x –1.26
Control method: Approved in vitro diagnostic(enzymatic method)
6. Standard Material
SRM911（NIST）＊

Storage and Shelf Life ＊＊1. Storage temperature: 2–10C
2. Shelf life: 18 months from the date ofmanufacture(The expiration date is printed on the outerpackage.)＊

Precautions for Use or Handling ＊1. 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) Cholestest N Calibrator and QUALIGENT N
Calibrator for LABOSPECT 008 containhuman-derived components that have beenshown to be negative for HBs antigens, HIVantibodies (AIDS virus antibodies), and HCV

2Packaging＊＊NameQUALIGENTCHO＊Package contents
CHO
EnzymeSolution 1Set
1 × 22.5 mL(Cassette forHitachi 9000
CHOseries)
EnzymeSolution 2
1 × 7.5 mL
CHO
EnzymeL set
Solution 1(Set for1 × 46 mL
HitachiCHO
LABOSPECT
Enzymeseries)
Solution 2
1 × 16 mL22

References ＊＊1) Sasaki M. et al.: Sampling of constituents of thehuman body, 246, Kodansha, 1972.
2) Kanai M. (supervising editor): Kanai’s manual ofclinical laboratory medicine. 34th ed. 512,
Kanehara Shuppan, 2015.
3) Japan Atherosclerosis Society, ed. Japan
Atherosclerosis Society (JAS) Guidelines forDiagnosis and Treatment of Atherosclerotic
Cardiovascular Diseases, 2002, 5.
4) 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