This package insert follows the Pharmaceuticals, Medical devices and Other Therapeutic Products Act of Japan.
In Vitro Diagnostics
Marketing Notification No.13A2X00197218043** Revised: January 2017 (8th edition)* Revised: September 2013 (7th edition)

This package insert must be read carefully prior to use.
LDL-cholesterol assay kit(Classification No.: 30173000)

correlated with ischemic heart disease than the totalcholesterol level.
The LDL-cholesterol concentration has been
calculated according to the Friedwald equation basedon measurement of 3 other variables (total cholesterol,HDL-cholesterol, and triglycerides). However, theaccuracy of this method is insufficient. Even thoughultracentrifugation has been also performed as thestandard method of measuring LDL-cholesterol, thismethod is difficult to use for laboratory tests inroutine clinical practice as requires special equipment
and time. The direct method, therefore, is widelyemployed in recent years.

QUALIGENT LDL
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.

Assay Principle
1. Assay Principle
The principle of this method is that eachlipoprotein reacts differently with surfactants
depending on its physicochemical properties.
Therefore, two different surfactants areemployed. Surfactant 1 is added in the firstreaction. It can change the structure oflipoproteins other than LDL, including
chylomicrons (CM), VLDL and HDL. In thepresence of this surfactant, lipoproteins otherthan LDL are eliminated by the action ofcholesterol oxidase and cholesterol esterase.
Surfactant 2, which promotes the enzymatic
reaction of all types of lipoproteins, is used in thesecond reaction. The enzymatic reaction with
LDL-cholesterol left from the first reaction isinitiated to produce color development.

Description (Kit Components) ＊＊Component : IngredientsLDL Enzyme Solution 1:
4-Aminoantipyrine
Cholesterol oxidase (microbialorigin)
Cholesterol esterase
Peroxidase
Surfactant 1
Good’s buffer
LDL Coloring Solution 2:
N,N-bis(4-sulfobutyl)-m-toluidine
disodium (DSBmT)
Surfactant 2
Good’s bufferFirst reaction:
HDL, VLDL, CM

Surfactant 1

Cholesterol esterase
Micellar cholesterol
H2O2
Cholesterol oxidase
H2O2 + 4-Aminoantipyrine
LDLLDL
Surfactant 1

Peroxidase
Colorless

Second reaction:
Intended Use
Measurement of LDL-cholesterol in serum orplasma

LDL

Surfactant 2

Cholesterol esterase
Micellar cholesterol
H2O2
Cholesterol oxidase

Cholesterol that exists in LDL (Low Density
Lipoprotein) fraction is called LDL-cholesterol.
Elevation of LDL-cholesterol is known to be a risk
factor for arteriosclerotic diseases, especiallycoronary artery disease.
Total cholesterol has commonly been measured for
diagnosis of hyperlipidemia, which is considered to
be a cause of arteriosclerotic diseases. However,according to the “Research Report 1986 on Specified
Disease Primary Hyperlipidemia by Investigation andResearch Group of the Ministry of Health andWelfare”1), the LDL-cholesterol level is more closely

Peroxidase
H2O2 + 4-Aminoantipyrine + DSBmTRed-purple color
2. Features
1) The direct method does not requirecomplicated procedures.
2) Results obtained by this method are closely
correlated with those by ultracentrifugation.
3) Analytical results are not affected by chyle (upto TG 1500 mg/dL).

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60–163 mg/dL
2. Diagnostic criterion4)
Hyper-LDL cholesterolemia: ≥ 140 mg/dL
3. There may be reactions with non-target substancesin the samples or interfering reactions. Especially
if abnormal lipid metabolism is suspected
including cases of biliary obstruction, accurate
measurements may not be obtained due to theinfluence of lipoprotein abnormalities. If assay
results appear to be unreliable, repeat themeasurement (if necessary, after dilution) or tryanother analytical methods.

＊ ＊Procedural Precautions ＊1. Properties of Samples and Sampling Methods
1) Samples
Serum and plasma (heparin plasma andEDTA plasma) may be used.
2) Storage of samples5)
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
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 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, use QUALIGENT N Calibrator for
LABOSPECT
008(manufacturedbySEKISUI MEDICAL) as the calibrationmaterial with a Hitachi LABOSPECT 008
Automated Analyzer.
2) Precautions for assay range
If the concentration of sample exceeds assayrange, dilute the sample with saline and repeat
the measurement.

Performance ＊＊1. Sensitivity
1) Reagent blank: absorbance being equal to or
lower than 0.05
2) Sensitivity :The absorbance is 0.18–0.28 per
100 mg/dL of LDL-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 Range5): (On a Hitachi 9000
series automated analyzer)
1–400 mg/dL
5. Correlation5)
1) Serum N=70 r=0.997 y0.96x＋0.02
Control method: Approved in vitro diagnostic(enzymatic assay)
2) Plasma N=60 r=0.999 y=1.03x –1.14
Control method: Approved in vitro diagnostic(enzymatic assay)
6. Standard Material
Certified Reference Material for Measurement ofHDL Cholesterol, LDL Cholesterol andTriglycerides in Human Serum (Reference
Material Institute for Clinical ChemistryStandards)＊ ＊Dosage/Administration (Assay Procedure) ＊1. Preparation of reagents
Reagent (1): LDL Enzyme Solution 1 is ready touse.
Reagent (2): LDL Coloring Solution 2 is ready touse.
2. Assay Procedure
This product is compatible with Hitachi 9000
series and LABOSPECT series automatedanalyzers. Assay procedure is indicated below.
SampleReagent (1)+2 L
180 LReagent (2)60 L37C5 min37C5 min＊ ＊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 HCVantibodies. 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 theLDL Enzyme Solution 1 andLDL Coloring
Solution 2. Therefore, if the reagent comes incontact with skin or clothes, rinse immediately
with ample water, and consult the doctor if skinirritation develops.
2. Precautions for use
1) This product should be stored as directed,without freezing. Freezing can deteriorate theMeasurement(Absorbance I※)
Measurement(Absorbance I※)
Calculation ofconcentration※

Absorbance I and II: The difference in absorbance
between 660 nm and 546 nm.
Calibration material: Cholestest N Calibrator
Use
QUALIGENTNCalibrator
forLABOSPECT 008 (manufactured by SEKISUIMEDICAL) as the calibration material with aHitachi LABOSPECT 008 Automated Analyzer.
(Manufacture's assigned value)
Reagent blank: Purified water or saline

Assessment of Assay Results ＊＊1. Reference standard range3)

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reagents, 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
4. Other precautions
1) Do not use the containers for other purposes.
2) Do not take apart the reagent cartridge before.

ContactSEKISUI MEDICAL CO., LTD.
international@sekisui.com
Manufacturer ＊＊SEKISUI MEDICAL CO., LTD.
1-3, Nihonbashi 2-chome, Chuo-ku, Tokyo, Japan

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 ＊＊Name

PackageSet(Cassette forHitachi 9000series)

QUALIGENTLDL
L set(Set for Hitachi
LABOSPECTseries)

LDL Enzyme
Solution 1
1 × 27.0 mL
LDL Coloring
Solution 2
1 × 9.0 mL
LDL Enzyme
Solution 1
1 × 48 mLLDL Coloring
Solution 2
1 × 16 mL×2×2

References ＊＊1) Tarui S.: Research Report 1986 on Specified
Disease Primary Hyperlipidemia by Investigation
and Research Group of the Ministry of Health andWelfare (group leader: Seiichi Tarui), 17–26,
1987.
2) Kanno T. et al.: J Med Pharm Sci, 37, 635–644,
1997.
3) Kanai M. (supervising editor): Kanai’s manual ofclinical laboratory medicine. 34th ed. 522,
Kanehara Shuppan, 2015.
4) Japan Atherosclerosis Society, ed. 2012 Japan
Atherosclerosis Society (JAS) Guidelines forPrevention of Atherosclerotic CardiovascularDiseases, 13.
5) In house data, SEKISUI MEDICAL CO., LTD.

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