This package insert follows the Pharmaceuticals, Medical devices and Other Therapeutic Products Act of Japan.
In Vitro Diagnostics
Marketing Notification No. 13A2X00197218050**Revised: January 2017 (13th edition)*Revised: March 2013 (12th edition)

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

converted to glycerol-3-phosphate by glycerol
kinase (GK) in the presence of adenosine
triphosphate (ATP). Then hydrogen peroxide is
produced by glycerol-3-phosphate oxidase (GPO),
after which hydrogen peroxide is decomposed
into water and oxygen by catalase and theseproducts are eliminated.
Second reaction: In the presence of LPL in TG-N
enzyme solution 2, triglycerides in the sample are
rapidly hydrolyzed to glycerol and fatty acids.
The glycerol thus formed is converted toglycerol-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
andN-ethyl-N-sulfobutyl-m-toluidine (ESBmT) in thepresence of peroxidase (POD) to form a
red-purple color. The triglyceride content isdetermined by measuring the absorbance of thesample. The influence of ascorbic acid is blocked
by ascorbate oxidase.

Pureauto S TG−NGeneral 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.

First reaction:
GK
Glycerol + ATP

Glycerol-3-phosphate + ADP

Glycerol-3-phosphate + O2 GPO
Dihydroxyacetone phosphate

Description (Kit Components) ＊＊Component: Ingredients
TG-N Enzyme Solution 1:
Glycerol kinase (microbial origin)
Glycerol-3-phosphate oxidase(microbial origin)
Catalase
Ascorbate oxidase
Adenosine triphosphate disodium
N-ethyl-N-sulfobutyl-m-toluidine
sodiumGood’s buffer
TG-N Enzyme Solution 2:
4-Aminoantipyrine
Lipoprotein lipase
Peroxidase
Good’s buffer

H2O2

Catalase

H2O2 +

H2O＋O2

Second reaction:
Triglycerides

LPL

Glycerol + Fatty acids

GK
Glycerol + ATP

Glycerol-3-phosphate + ADP

Glycerol-3-phosphate + O2Dihydroxyacetone phosphate
H2O2 + ESBmTRed-purple color+GPO

H2O2 +4-Aminoantipyrine

POD

2. Features
1) Assay results are hardly affected by crosscontamination.
2) Free glycerol is eliminated by the enzymaticmethod.
3) Liquid reagents, ready-to-use.
4) Wide assay range.
5) Bilirubin, hemolysis, ascorbic acid and chylehave minimal effects on results.
6) Applicable to various automated analyzers.

Intended Use
Measurement of triglyceride in serum
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 may be used.
2) Storage of samples1)

Assay Principle
1. Assay Principle
First reaction: Free glycerol in the sample is1

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.
3) Measurement of heparinized samples may
yield artificially low values because
decomposition of triglycerides is promoted.4)
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.

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-housemethods.)
4. Measurement Range5): (On Hitachi 7170S
automated analyzer)
3–2000 mg/dL
5. Correlation5)
Serum N=55 r=0.999 y=1.00x–1.0
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
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 contains human-derived
components that have been 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) TG-N Enzyme Solution 2 contains sodium
azide to inhibit catalase activity. If this solutionis accidentally ingested or comes into contact
with the eyes or skin, immediately rinse the areawith water and seek medical treatment, ifnecessary.
4) Proclin 300, which possesses skin-irritative
potential, is added as an antiseptic agent in theTG-N Enzyme Solution 2. Therefore, if thereagent 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 Disposal1) 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.

Dosage/Administration (Assay Procedure) ＊＊1. Preparation of reagents
Reagent (1): TG-N Enzyme Solution 1 is readyto use.
Reagent (2): TG-N Enzyme Solution 2 is readyto use.
2. Assay Procedure
This product is compatible with various types ofautomated analyzer. An example of the assay
procedure is indicated below.
Sample+3 LReagent(1)
260 μL
Reagent(2)
130 μL

37C
5 min
37C
5 min

Measurement(Absorbance I※)
Measurement(Absorbance II※)
Calculate 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)
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 or
lower than 0.052) Sensitivity: The absorbance is 0.096–0.162）To prevent infections from spilled samples orsolutions containing samples, wipe the spilledarea thoroughly with disinfectants such as
sodium hypochlorite solution at a concentration2

of 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) TG-N Enzyme Solution 2 contains sodium
azide to inhibit catalase activity. Sodium azide
may react with lead or copper pipes andproduce highly explosive metallic azide. Whendisposing of this product, flush it away withcopious 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
TG-N Enzyme(1)
2 × 400 mL
Solution 1
Pureauto S
TG−N
TG-N Enzyme(2)
2 × 200 mL
Solution 2
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) Shirai K.: Modern Med Lab., 21, 3, 211, 1993.
5) 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