This package insert follows the Pharmaceuticals, Medical devices and Other Therapeutic Products Act of Japan.

In Vitro Diagnostics **Revised: January 2017 (12th edition)

Marketing Notification No. 13A2X00197218044 *Revised: September 2016 (11th edition)
This package insert must be read carefully prior to use.

LDL-cholesterol assay kit standard method of measuring LDL-cholesterol, this

(Classification No.: 30173000) method is difficult to use for laboratory tests in
routine clinical practice as requires special equipment
and time. The direct method, therefore, is widely
Cholestest LDL employed in recent years.

General Precautions Assay Principle

1. This product is for in vitro diagnostic use, and 1. Assay Principle
must not be used for any other purposes. The principle of this method is that each
2. Clinicians should make a comprehensive clinical lipoprotein reacts differently with surfactants
decision based on assay results in conjunction with depending on its physicochemical properties.
clinical symptoms and other examination results. Therefore, two different surfactants are employed.
3. This product should be used only as directed in this Surfactant 1 is added in the first reaction. It can
package insert. Reliability of results cannot be change the structure of lipoproteins other than
guaranteed if there are any deviations from the LDL, including chylomicron (CM), very VLDL
instructions in this package insert. and HDL. In the presence of this surfactant,
4. If reagent accidentally comes in contact with eyes lipoproteins other than LDL are eliminated by the
and/or mouth, rinse immediately with ample water action of cholesterol oxidase and cholesterol
as first aid, and consult the doctor if required. esterase. Surfactant 2, which promotes the
5. Carefully read the operating instructions for each enzymatic reaction of all types of lipoproteins, is
type of automated analyzers prior to using this used in the second reaction. The enzymatic
product. Parameters for each type of analyzers are reaction with LDL-cholesterol left from the first
available, and can be requested from SEKISUI reaction is initiated to produce color development.
MEDICAL CO., LTD. if required.
6. Perform a quality control test prior to assay to First reaction:
ensure accuracy. Surfactant 1
HDL, VLDL, CM
Description (Kit Components) ＊ Cholesterol esterase
Component: Ingredients Micellar cholesterol H2O2
Enzyme Solution: 4-Aminoantipyrine Cholesterol oxidase
Cholesterol oxidase
Peroxidase
Cholesterol esterase H2O2 + 4-Aminoantipyrine Colorless
Peroxidase
Coloring Solution: N,N-bis LDL Surfactant 1 LDL
(4-sulfobutyl)-m-toluidine
disodium (DSBmT) Second reaction:
Intended Use Surfactant 2
LDL
Measurement of LDL-cholesterol in serum or
plasma 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 Peroxidase
H2O2 + 4-Aminoantipyrine + DSBmT
factor for arteriosclerotic diseases, especially Red-purple color
coronary artery disease. Total cholesterol has
commonly been measured for diagnosis of 2. Features
hyperlipidemia, which is considered to be a cause of 1) The direct method does not require
arteriosclerotic diseases. However, according to the complicated procedures
“Research Report 1986 on Specified Disease Primary 2) Results obtained by this method are closely
Hyperlipidemia by Investigation and Research Group correlated with those by ultracentrifugation.
of the Ministry of Health and Welfare”1), the 3) Analytical results are not affected by chyle (up
LDL-cholesterol level is more closely correlated with to TG 1500 mg/dL).
ischemic heart disease than the total cholesterol level.
The LDL-cholesterol concentration has been Procedural Precautions ＊
calculated according to the Friedwald equation based 1. Properties of Samples and Sampling Methods
on measurement of 3 other variables (total cholesterol, 1) Samples
HDL-cholesterol, and triglycerides). However, the Serum and plasma may be used.
accuracy of this method is insufficient. Even though 2) Storage of samples4)
ultracentrifugation has been also performed as the If the isolated serum or plasma sample cannot
1
 be tested on the same day, specimens should be 100 mg/dL of LDL-cholesterol.
stored as follows: 2. Accuracy: 90–110% of the expected assay value
2–10C: for tests within 1 week 3. Within-run Reproducibility:
≤ -20C: for tests after more than 1 week Coefficient of variation ≤ 5%
Do not refreeze. (Test methods used for 1. –3. are in-house
Bring samples to room temperature (15–30C) methods.)
before use. 4. Measurement Range4):(On Hitachi 7150
2. Interfering substances automated analyzer)
Assay results are not affected by free bilirubin (up 1–450 mg/dL
to 20 mg/dL), conjugated bilirubin (up to 20 5. Correlation4)
mg/dL), hemoglobin (up to 500 mg/dL), ascorbic 1) Serum N=60 r=0.992 y=0.95x＋3.66
acid (up to 50 mg/dL), formazin turbidity (up to Control method: Ultracentrifugation
2500 FTU), or Intralipos (up to 5%). 2) Plasma N=76 r=0.998 y=1.01x –4.03
3. Others Control method: Comparison with the values
1) Always use Cholestest N Calibrator or for plasma samples obtained simultaneously
QUALIGENT N Calibrator for calibration. with the serum samples.
2) Precautions for assay range 6. Standard Material
If the concentration of sample exceeds assay Certified Reference Material for Measurement of
range, dilute the sample with saline and repeat HDL Cholesterol, LDL Cholesterol and
the measurement. Triglycerides in Human Serum (Reference
Material Institute for Clinical Chemistry
Dosage/Administration (Assay Procedure) ＊ Standards)
1. Preparation of reagents
Reagent (1): Enzyme Solution is ready to use. Precautions for Use or Handling ＊
Reagent (2): Coloring Solution is ready to use. 1. Precautions for Handling (to Ensure Safety)
2. Assay Procedure 1) All samples used in the test should be handled
This product is compatible with various types of as a material possibly infected with HIV, HBV,
automated analyzer. An example of the assay HCV, or other viruses. To prevent infection, use
procedure is indicated below. disposable gloves and avoid mouth pipetting
during the test.
37C Measurement 2) Cholestest N Calibrator and QUALIGENT N
Sample Reagent (1)
+ Calibrator for LABOSPECT 008 contain
3 μL 300 μL 5 min (Absorbance I※)
human-derived components that have been
37C Measurement
Reagent (2) shown to be negative for HBs antigens, HIV
100 μL 5 min (Absorbance II※) antibodies (AIDS virus antibodies), and HCV
Calculation of antibodies. However, these reagents (as well as
concentration the samples) should be considered potentially
infectious and handled with great care.
※
Absorbance I and II: The difference in absorbance 3) Proclin 300, which possesses skin-irritative
between 660 nm and 546 nm. potential, is added as an antiseptic agent in the
Calibration material: Cholestest N Calibrator Enzyme Solution and Coloring Solution.
(in-house indicated values) or QUALIGENT N Therefore, if the reagent comes in contact with
Calibrator for LABOSPECT 008 (Manufacture's skin or clothes, rinse immediately with ample
assigned value) water, and consult the doctor if skin irritation
Reagent blank: Purified water or saline develops.
2. Precautions for use
Assessment of Assay Results ＊＊ 1) This product should be stored as directed,
1. Reference standard range2) without freezing. Freezing can deteriorate the
65–163 mg/dL reagents, which can produce inaccurate results.
2. Diagnostic criterion3) Therefore, avoid using the reagents which have
Hyper-LDL cholesterolemia: ≥ 140 mg/dL been previously frozen.
3. There may be reactions with non-target substances 2) Do not use expired reagents. Use of such
in the samples or interfering reactions. Especially reagents cannot guarantee the reliability of
if abnormal lipid metabolism is suspected, measurement values.
including cases of biliary obstruction, accurate 3) Do not replenish the reagents.
measurements may not be obtained due to the 4) Do not perform the assay under direct sunlight
influence of lipoprotein abnormalities. If assay 3. Precautions for Disposal
results appear to be unreliable, repeat the 1) Before disposal, used samples and their
measurement (if necessary, after dilution) or try containers must be immersed in sodium
another analytical methods. hypochlorite solution at a concentration of
greater than 0.1% for longer than 1 hour or
Performance autoclaved at 121ºC for 20 minutes.
1. Sensitivity 2）To prevent infections from spilled samples or
1) Reagent blank: absorbance being equal to or solutions containing samples, wipe the spilled
lower than 0.05 area thoroughly with disinfectants such as
2) Sensitivity: The absorbance is 0.18–0.28 per sodium hypochlorite solution at a concentration
2
 of greater than 0.1%.
3) The reagents and treated samples should be
discarded as medical waste or industrial waste
according to the waste disposal regulations.
4) The reagents should be disposed of in
accordance with the Water Pollution Control act
or related regulations.
4. Other precautions
Do not use the containers for other purposes.

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 outer
package.)
Packaging

Name Package
Cholestest (1) Enzyme Solution 2 × 6 0mL
LDL (2) Coloring Solution 2 × 20 mL
Reagents are available in other configurations. For
further details please contact SEKISUI MEDICAL
CO., LTD.

References ＊＊
1) Tarui S.: Research Report 1986 on Specified
Disease Primary Hyperlipidemia by Investigation
and Research Group of the Ministry of Health and
Welfare (group leader: Seiichi Tarui), 17–26,
1987.
2) Kanai M. (supervising editor): Kanai’s manual of
clinical laboratory medicine. 34th ed. 522,
Kanehara Shuppan, 2015.
3) Japan Atherosclerosis Society, ed. 2012 Japan
Atherosclerosis Society (JAS) Guidelines for
Prevention of Atherosclerotic Cardiovascular
Diseases, 13.
4) In house data, SEKISUI MEDICAL CO., LTD.

Contact
SEKISUI MEDICAL CO., LTD.
[email protected]

Manufacturer ＊＊
SEKISUI MEDICAL CO., LTD.
1-3, Nihonbashi 2-chome, Chuo-ku, Tokyo, Japan