This package insert follows the Pharmaceuticals, Medical devices and Other Therapeutic Products Act of Japan.
In Vitro Diagnostics
Marketing Notification No. 13A2X00197218047**Revised: January 2017 (9th edition)*Revised: April 2008 (8th edition)

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

cholesterol oxidase (COD) to form hydrogen
peroxide. Hydrogen peroxide causes oxidative
condensationof
4-aminoantipyrine
andN-ethyl-N-sulfobutyl-m-toluidine (ESBmT) in thepresence of peroxidase (POD) to form a
red-purple color. The total cholesterol content isdetermined by measuring the absorbance. The
influence of ascorbic acid is blocked by ascorbate
oxidase.
CEEsterified cholesterol ─▶ Free cholesterol + Fatty
acid
CODFree cholesterol ─▶ Δ4-cholestenone + H2O2
POD
H2O2 + ESBmT + 4-Aminoantipyrine ─▶
Red-purple color
2. Features
1) Liquid reagents, ready-to-use.
2) Wide assay range.
3) Assay results are hardly affected by crosscontamination.
4) Bilirubin, hemolysis, ascorbic acid and chylehave minimal effects on results.
5) Applicable to various automated analyzers.

Pureauto S CHO−N
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.

＊ ＊Procedural Precautions ＊1. Properties of Samples and Sampling Methods
1) Samples
Serum may be used.
2) Storage of samples1)
If the isolated serum sample cannot be testedon 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), andascorbic acid (50 mg/dL).
3. Others
1) Always use Seronorm Lipid or Cholestest N
Calibrator for calibration.
2) Precautions for assay range
If the concentration of sample exceeds assay
range, dilute the sample with saline and repeat
the measurement.

Description (Kit Components) ＊＊Component: Ingredients
CHO-N Enzyme Solution 1:
4-Aminoantipyrine
Cholesterol esterase
Peroxidase
Ascorbate oxidase
Phthalate buffer
CHO-N Enzyme Solution 2:
Cholesterol oxidase (microbialorigin)
N-ethyl-N-sulfobutyl-m-toluidine
sodiumPhthalate buffer
Intended Use
Measurement of total cholesterol in serum
Cholesterol in serum is derived from dietary intake
and is also synthesized in the liver. Esterified and freecholesterol account for approximately 70% and 30%of total cholesterol, respectively.
Measurement of cholesterol is important as an index
of abnormal lipid metabolism. The esterified/freecholesterol ratio is also considered to be an important
index of severe parenchymal liver disease.

Dosage/Administration (Assay Procedure) ＊＊1. Preparation of reagents
Reagent (1): CHO-N Enzyme Solution 1 isready to use.
Reagent (2): CHO-N Enzyme Solution 2 isready to use.
2. Assay Procedure
This product is compatible with various types ofautomated analyzer. An example of the assay

Assay Principle
1. Assay Principle
Cholesterol exists in samples as both the esterified
and free types. Esterified cholesterol is converted
to free cholesterol by action of cholesterolesterase (CE). Then free cholesterol is oxidized by1

procedure is indicated below.
Sample + Reagent 1
3 μL
260 μL
Reagent 2
130 μLreagent comes in contact with skin or clothes,rinse immediately with ample water, and consult
the doctor if skin irritation develops.
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 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 has been added as an antiseptic
agent in the CHO-N Enzyme Solution 2. It canreact with lead or copper pipes to produce thehighly explosive metal azide. Therefore, thereagent should be flushed with large amounts ofwater during disposal.
4. Other precautions
Do not use the containers for other purposes.

37C Measurement(Absorbance I※)
5 min
37C Measurement(Absorbance II※)
5 min
Calculation ofconcentration※

Absorbance I and II: The difference in absorbance
between 700 nm and 600 nm.
Calibration material: Seronorm Lipid or
Cholestest N Calibrator (Manufacture's assignedvalue)
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 0.05
2) Sensitivity: The absorbance is 0.34–0.47 per
300 mg/dL of cholesterol.
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 Range4): (On Hitachi 7170S
automated analyzer)
4–1300 mg/dL
5. Correlation4)
1) Serum N=100 r=0.999 y=1.01x–3.85
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 outer
package.)
Packaging
Name
Package
CHO-N Enzyme(1)
2 × 400 mL
Solution 1
Pureauto S
CHO−N
CHO-N Enzyme(2)2 × 200 mL
Solution 2
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)
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) Sodium azide is added as an antiseptic agent inthe CHO-N Enzyme Solution 2. Therefore, if
the reagent comes in accidentally contact witheyes, mouth or skin, rinse immediately withample water as first aid, and consult the doctor
if required.
3) Proclin 300, which possesses skin-irritative
potential, is added as an antiseptic agent in theCHO-N Enzyme Solution 1. Therefore, if the＊ ＊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. 2002 Japan

2Atherosclerosis Society (JAS) Guidelines forPrevention of Atherosclerotic CardiovascularDiseases, 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