This package insert follows the Pharmaceuticals, Medical devices and Other Therapeutic Products Act of Japan.
In Vitro Diagnostics
Marketing Notification No.13A2X00197218042**Revised: January 2017 (13th edition)*Revised: September 2016 (12th edition)

This package insert must be read carefully prior to use.
HDL-cholesterol assay kit(Classification No.: 30169000)

surfactant with different effects on HDL and otherlipoproteins (LDL, VLDL and chylomicron).
Therefore, only HDL-cholesterol is specificallymeasured.
Surfactant
HDL, LDL, VLDL, Chylomicron
HDL issolubilized
Cholesterol esterase
HDL-cholesterol
Δ4-cholestenone + H2O2
Cholesterol oxidase
Peroxidase
H2O2 + DSBmT + 4-Aminoantipyrine
Red-purple color

Cholestest N HDLGeneral 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.

2. Features
1) This product is used to measure
HDL-cholesterol by the direct method.
2) Because it does not contain Mg salt, there is
no heavy burden imposed on analyzer.
Procedural Precautions ＊1. Properties of Samples and Sampling Methods
1) Samples
Serum and plasma (heparin plasma andEDTA plasma) may be used.
2) Storage of samples3)4)
If a serum or plasma sample cannot bemeasured on the day of separation, store thesample as follows:
2–10C: for tests within 1 week≤- 20C: for tests after more than 1 week
Do not re-freeze.
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 50 mg/dL), or Intralipos (up to 5%).
3. Others
1) Always use Cholestest N Calibrator or
QUALIGENT 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
Enzyme Solution 1: N,N-bis(4-sulfobutyl)-m-toluidine
disodium (DSBmT)
Cholesterol oxidase (microbialorigin)
Peroxidase
Enzyme Solution 2: 4-Aminoantipyrine
Cholesterol esterase
Surfactant
Intended Use
Measurement of HDL-cholesterol in serum orplasma
Cholesterol that exists in HDL (High Density
Lipoprotein) fraction is called HDL-cholesterol. The
Framingham study (1977) and other studies haveshown that a low level of HDL-cholesterol is a risk
factor for ischemic heart disease.1
HDL-cholesterol is decreased in patients withcoronary artery disease, hyperlipidemia, smoking,obesity, diabetes, and hepatic dysfunction, while it isincreased by alcohol intake and by adequate exercise.
The HDL-cholesterol concentration also varies
depending on age, gender, and hereditary factors.

Dosage/Administration (Assay Procedure)
1. Preparation of reagents
Reagent (1): Enzyme Solution 1 is ready to use.
Reagent (2): Enzyme Solution 2 is ready to use.
2. Assay Procedure
This product is compatible with various types ofautomated analyzer. An example of the assay
procedure is indicated below.

Assay Principle
1. Assay Principle
In this assay system, HDL-cholesterol isselectively solubilized, and rapidly undergoes anenzymatic reaction due to the use of special

12) Cholestest N Calibrator and QUALIGENT N
Calibrator for LABOSPECT 008 containhuman-derived components
shown to be
negative for HBs antigens, HIV antibodies(AIDS virus antibodies), and HCV antibodies.
However, these reagents (as well as thesamples) 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 theEnzyme Solution 1 and Enzyme Solution 2.
Therefore, if the reagent comes in contact withskin 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.
4. Other precautions
Do not use the containers for other purposes.

37C Measurement
Sample Reagent (1)+───▶
3 L(Absorbance I※)
300 μL
5 min
37C Measurement
Reagent (2)───▶ (Absorbance II※)
100 μL
5 min
Calculation of────▶
concentration※

Absorbance I and II: The difference in absorbance
between 700 nm and 600 nm.
Calibration material: Cholestest N Calibrator(in-house indicated values) or QUALIGENT N
Calibrator for LABOSPECT 008 (Manufacture'sassigned value)
Reagent blank: Purified water or saline

Assessment of Assay Results ＊＊1. Reference standard range1)
Male: 38–90 mg/dL
Female: 48–103 mg/dL
2. Diagnostic criterion2)
Low-HDL cholesterolemia: < 40 mg/dL
3. There may be reactions with non-target substancesin the samples or interfering reactions. If assay
results appear to be unreliable, repeat themeasurement (if necessary, after dilution) or try
another analytical methods.
Performance
1. Sensitivity1) Reagent blank: absorbance being equal to or
lower than 0.05
2) Sensitivity: 0.07–0.17 per 100 mg/dL ofHDL-cholesterol.
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 Range4): (On Hitachi 7170
automated analyzer)
2–150 mg/dL
5. Correlation4)
1) Serum N=56 r=0.999 y=0.99x+0.39
Control method: Approved in vitro diagnostic(selective inhibition assay)
2) Plasma N=76 r=0.996 y=0.97x +0.28
Comparison with the values for plasma samples
obtained simultaneously with the serumsamples.
6. Standard Material
Certified Reference Material for Measurement ofHDL Cholesterol, LDL Cholesterol andTriglycerides in Human Serum (Reference
Material Institute for Clinical ChemistryStandards)

Storage and Shelf Life
1. Storage temperature: 2–10C
2. Shelf life: 2 years from the date of manufacture(The expiration date is printed on the outerpackage.)
Packaging
NamePackage
Enzyme Solution 1 2 × 60 mL
Cholestest NHDL
Enzyme Solution 2 2 × 20 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)
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.

References ＊＊1) Kanai M. (supervising editor): Kanai’s manual ofclinical laboratory medicine. 34th ed. 520,
Kanehara Shuppan, 2015.

22) Japan Atherosclerosis Society, ed. Japan
Atherosclerosis Society (JAS) Guidelines for
Diagnosis and Treatment of Atherosclerotic
Cardiovascular Diseases, 2002, 5.
3) Kodajima N. et al.: J Med Pharm Sci, 46, 235,2001.
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, Japan

This product uses a patent related to HDL-C
measuring reagent and method (patent No. 3288033),
and patent application (PCT/JP00/03860).

3