This package insert follows the Pharmaceuticals, Medical devices and Other Therapeutic Products Act of Japan.
In Vitro Diagnostics
Marketing Notification No. 13A2X00197218051**Revised: December 2017 (16th edition)*Revised: January 2017 (15th edition)

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

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 these products
are eliminated.
Second reaction: In the presence of lipoprotein
lipase (LPL) in TG-N Reagent 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-sulfobutyl-mtoluidine (ESBmT) in the presence of peroxidase(POD) to form a red-purple dye. The triglyceride
content is determined by measuring the absorbanceof the red-purple dye. The influence of ascorbic
acid is blocked by ascorbate oxidase.

Autosera S TG-N
General 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 be
guaranteed 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

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

H2O2

CatalaseGPO

H2O2 +H2O + O2

Second reaction:
Triglycerides

LPL

Glycerol + ATP

GK

Glycerol + Fatty acids
Glycerol-3-phosphate + ADP

GPO

Glycerol-3-phosphate + O2
Dihydroxyacetone phosphate

H2O2 +H2O2 + ESBmT + 4-Aminoantipyrine
purple color

POD

Red-2. Features
1) With this product, assay results are hardly
affected even if cross-contamination occurs.
2) Free glycerol is eliminated by the enzymatic
method.
3) Liquid reagents, ready-to-use.
4) Wide assay range.
5) Bilirubin, hemolysis, ascorbic acid and chylehave minimal effects on results.

Intended Use
Measurement of triglycerides 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)
If the isolated serum sample cannot be testedon the same day, specimens should be stored asfollows:
2–10C: for tests within 1 week≤ -20C: for tests after more than 1 week

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

15. Correlation5)
Serum
N=50
r=0.999
y=1.01x+0.02
Control method: Approved in vitro diagnostic(enzymatic method)
6. Standard Material
Reference standard for lipid assay (ReferenceMaterial Institute for Clinical ChemistryStandards)

Bring samples to room temperature (15–30C)
before use.
3) Measurement of heparinized samples may yieldartificially low values because decompositionof 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), orascorbic 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.

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 contains human-derived
components determined as HBsAg-negative,
HIV antibody (AIDS virus antibody) negative,and HCV antibody negative. When using,
however, it should be handled very carefully aswith samples, considering the risk of infectious.
3) TG-N Reagent 2 contains sodium azide to inhibit
catalase activity. If this substance is accidentally
ingested or comes into contact with the eyes orskin, immediately implement first-aid measures
such as rinsing the area with water and seek
medical treatment if necessary.
4) Proclin 300, which possesses skin-irritative
potential, is added as an antiseptic agent in theTG-N Reagent 2. Therefore, if the reagent comesin contact 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 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.

Dosage/Administration (Assay Procedure) ＊1. Preparation of reagents
Reagent (1): TG-N Reagent 1 is ready to use.
Reagent (2): TG-N Reagent 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.
37C Measurement
Sample + Reagent (1)(Absorbance I※)
3 μL
260 μL
5 min
Reagent (2) 37C Measurement(Absorbance II※)
130 μL
5 min
Calculation 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 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 or
lower than 0.05
2) Sensitivity: The absorbance is 0.096–0.16 per200 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 Range5): (On Hitachi 7170S
automated analyzer)
3–2000 mg /dL5) TG-N (2) contains sodium azide to inhibit
catalase activity. Sodium azide may react withlead or copper pipes and produce highly2

explosive metallic azide. When disposing of thisproduct, flush it away with copious amounts ofwater.
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 Reagent 1
4 × 85 mL(1)
TG-N Reagent 1
4 × 50 mL
Autosera S
TG-N
TG-N Reagent 2
4 × 45mL(2)
TG-N Reagent 2
4 × 50 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 Guidelines for Preventionof 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, Japan

3