This package insert follows the Pharmaceuticals, Medical devices and Other Therapeutic Products Act of Japan.
In Vitro Diagnostics
Marketing Notification No. 13A2X00197218049**Revised: January 2017 (11th edition)*Revised: September 2016 (10th edition)

This package insert must be read carefully prior to use.
Triglyceride assay kit(Classification No.: 30182000)

these products are eliminated.
Second reaction: In the presence of lipoprotein
lipase (LPL) in Enzyme Solution 2, triglycerides inthe sample are rapidly hydrolyzed to glycerol andfatty acids. The glycerol thus formed is converted
to glycerol-3-phosphate by the action of GK in thepresence of ATP, which is then converted tohydrogen peroxide in the presence of GPO.
Hydrogen peroxide causes oxidative condensationof 4-aminoantipyrine and N-Ethyl-N-sulfobutylm-toluidine sodium (ESBmT) in the presence ofperoxidase (POD) to form a red-purple dye. The
triglyceride content is determined by measuring
the absorbance of the red-purple dye. The
influence of ascorbic acid is blocked by ascorbate
oxidase.

Cholestest TGGeneral Precautions
1. This product is for in vitro diagnostic use, and mustnot 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 if required.
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.

First reaction:
Glycerol + ATP

GK

Glycerol-3-phosphate + ADP

Glycerol-3-phosphate + O2
Dihydroxyacetone phosphate

H2O2Catalase

GPO

H2O2 +

H2O + O2

Second reaction:

Description (Kit Components) ＊Component: Ingredients
Enzyme Solution 1:
Glycerol kinase (microbial origin)
Glycerol-3-phosphate oxidase(microbial origin)
N-Ethyl-N-sulfobutyl-m-toluidine
sodiumEnzyme Solution 2:
4-Aminoantipyrine
Lipoprotein lipase
Peroxidase

Triglycerides

LPLGlycerol + ATPGK

Glycerol + Fatty acids
Glycerol-3-phosphate + ADP

Glycerol-3-phosphate + O2
Dihydroxyacetone phosphateGPO

H2O2 + ESBmT + 4-Aminoantipyrine

H2O2 +POD
Red-purple color

2. Features
1) With this reagent, measurements of TG and otherparameters (such as F-CHO, NEFA) is hardly
affected even if cross-contamination occurs.
2) Free glycerol is eliminated by the enzymaticmethod.
3) Bilirubin, hemolysis, ascorbic acid and chyle haveminimal effects on results.
4) Applicable to various automated analyzers.

Intended Use
Measurement of triglycerides in serum or plasma
Triglycerides are formed by esterification between 3
fatty acid molecules and 1 glycerol molecule, and are
the major component of lipids throughout the body.
Measurement of triglycerides is considered to be
useful for elucidating abnormalities of lipid
metabolism. Attention has especially been focused on
the association of triglycerides with arteriosclerosis
and coronary artery disease.

Procedural Precautions ＊1. Properties of Samples and Sampling Methods
1) Samples
Serum and plasma (heparin plasma and EDTAplasma) 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.
3) Measurement of heparinized samples may yield

Assay Principle
1. Assay Principle
First reaction: Free glycerol in the sample isconverted to glycerol-3-phosphate by glycerol
kinase (GK) in the presence of adenosine
triphosphate disodium (ATP). Then hydrogen
peroxide is produced by glycerol-3-phosphate
oxidase (GPO), after which hydrogen peroxide isdecomposed into water and oxygen by catalase and1artificially low values because decomposition oftriglycerides is promoted. 5)
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), andascorbic acid (50 mg/dL).
3. Others
1) Always use Seronorm Lipid, Cholestest NCalibrator
or QUALIGENT N Calibrator for
Labospect 008 for calibration.
2) Precautions for assay range
If the concentration of sample exceeds assay range,
dilute the sample with saline and repeat themeasurement.

automated analyzer)
3–2000 mg/dL
5. Correlation6)
1) Serum N=60
r=0.999
y=1.04x–0.14
Control method: Approved in vitro diagnostic(enzymatic method)
2) Plasma N=71
r=0.999
y=0.99x–0.46
Control method: Approved in vitro diagnostic(enzymatic method)
6. Standard Material
Reference standard for lipid assay (Reference
Material Institute for Clinical ChemistryStandards)
Precautions for Use or Handling ＊1. Precautions for Handling (to Ensure Safety)
1) All samples used in the test should be handled asa 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 contains humanderived components determined as HBsAgnegative, HIV antibody (AIDS virus antibody)
negative, and HCV antibody negative. Whenusing, however, it should be handled very
carefully as with samples, considering the risk ofinfectious.
3) Enzyme Solution 2 contains 0.0005 w/v%
sodium azide to inhibit catalase activity. If thissolution is accidentally ingested or comes into
contact with the eyes or skin, immediately
implement first-aid measures such as rinsing thearea with water and seek medical treatment ifnecessary.
4) Proclin 300, which possesses skin-irritative
potential, is added as an antiseptic agent in theEnzyme Solution 2. Therefore, if the reagent
comes in contact with skin or clothes, rinse
immediately with ample water, and consult thedoctor 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 orautoclaved 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

Dosage/Administration (Assay Procedure) ＊1. Preparation of reagents
Reagent (1): TG Enzyme Solution 1 is ready touse.
Reagent (2): TG Enzyme Solution 2 is ready touse.
2. Assay Procedure
This product is compatible with various types ofautomated analyzer. An example of the assay
procedure is indicated below.
37C
Sample + Reagent (1)2.4 μL240 μL
5 min
Reagent (2) 37C80 μL

5 min

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

Absorbance I and II: The difference in absorbance
between 800 nm and 600 nm.
Calibration material: Seronorm Lipid, Cholestest
N Calibrator (manufacture's assigned value), orQUALIGENT N Calibrator for Labospect 008(manufacture's assigned value)
Reagent blank: Purified water or saline

Assessment of Assay Results ＊＊1. Reference standard range2)
Male:
40–234 mg/dL
Female: 30–117 mg/dL
2. Diagnostic criterion3)
Hypertriglyceridemia: ≥ 150 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 orlower than 0.05
2) Sensitivity: The absorbance is 0.09–0.17 per
200 mg/dL of triglycerides.
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-house methods.)
4. Measurement Range6): (On Hitachi 7150

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according to the waste disposal regulations.
4) The reagents should be disposed of inaccordance with the Water Pollution Control act
or related regulations.
5) Enzyme Solution 2 contains 0.0005 w/v%
sodium azide, which inhibits the activity ofcatalase. Sodium azide may react with lead orcopper pipes and produce highly explosive
metallic azide. When disposing of this product,flush it away with copious amounts of water.
4. Other precautions
Do not use the containers for other purposes.
Storage and Shelf Life
1. Storage temperature: 2–10 C
2. Shelf life: 1 year from the date of manufacture(The expiration date is printed on the outerpackage.)
Packaging
NamePackage
Cholestest (1) Enzyme Solution 1 2 × 400 mLTG(2) Enzyme Solution 2 2 × 200 mL
Constituent reagents are available in otherconfigurations. For further details please contact
SEKISUI MEDICAL CO., LTD.
References ＊＊1) Sasaki M. et al.: Sampling of constituents of thehuman body, 263, Kodansha, 1972.
2) Kanai M. (supervising editor): Kanai’s manual ofclinical laboratory medicine. 34th ed. 514, Kanehara
Shuppan, 2015.
3) Japan Atherosclerosis Society, ed. 2012 Japan
Atherosclerosis Society (JAS) Guidelines forPrevention of Atherosclerotic Cardiovascular
Diseases, 13.
4) Tamura K. et al.: J Med Pharm Sci, 49, 791, 2003.
5) Shirai K.: Modern Med Lab, 21, 3, 211, 1993.
6) 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

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