This package insert follows the Pharmaceuticals, Medical devices and Other Therapeutic Products Act of Japan.
In Vitro Diagnostics
Marketing Approval No. 20300AMZ00140000** Revised: January 2017 (8th edition)* Revised: April 2008 (7th edition)

This package insert must be read carefully prior to use.
withdiabetes,
hyperthyroidism.

Class II Biochemical Test Series（General）(Classification No.: 80022002)
Nonesterified fatty acid assay kit(Classification No.: 30174000)

severehepatopathy,

andAssay Principle
1. Assay Principle
Free fatty acids in samples are converted to acyl
CoA by the action of CoA synthetase (ACS) in
the presence of coenzyme A (CoA) and adenosinetriphosphate (ATP). Acyl CoA is oxidized by acyl
CoA oxidase (ACO) to produce hydrogenperoxide. Hydrogen peroxide causes oxidative
condensation of 4-aminoantipyrine and N,N,-bis(4-sulfobutyl)-m-toluidine (DSBmT) in thepresence of peroxidase (POD) and form acomplex with a red-purple color. The content of
free fatty acids is determined by measuring theabsorbance of this complex.

Clinimate NEFA
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 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.

ACSFree fatty acids + CoA + ATP
2+Mg+ AMP + Pyrophosphoric acid

Acyl CoAACO
AcylCoA+O2
2,3-Trans-enoyl CoA + H2O2
POD
H2O2 + DSBmT + 4-Aminoantipyrine
Red-purple color
2. Features
This reagent is stable after preparation.
Procedural Precautions
1. Properties of Samples and Sampling Methods1) Samples
Serum may be used.
2) Storage of samples
The isolated serum should be tested on thesame day.
2. Interfering substances
1) Assay results are found to be insensitive bybilirubin (up to 5 mg/dL) and ascorbic acid(up to 50 mg/dL).
2) Hemolysis samples may result in positive
error. Perform re-measurement by another test
method if the results obtained are extremely
high compared with those of other related
variables.
3. Others
1) Always use Anaserum NEFA standard solution
for calibration.
2) Precautions for assay range
If the concentration of sample exceeds assay
range, dilute the sample with saline and repeat
the measurement.

Description (Kit Components) ＊＊Component: Ingredients
NEFA Enzyme Agent 1:
Acyl CoA synthetase (Pseudomonasorigin)
4-Aminoantipyrine
Ascorbate oxidase
Coenzyme A
Adenosine triphosphate disodium
Magnesium chloride
NEFA Buffer Solution 1:
2-Amino-2-hydroxymethyl-1,3-propanediol buffer
NEFA Enzyme Agent 2:
Acyl CoA oxidase (Arthrobacterorigin)
Peroxidase
NEFA Buffer Solution 2:
N, N-bis (4-sulfobutyl)-m-toluidine
disodium (DSBmT)N-Ethyl maleimide
Intended Use
Measurement of free fatty acids in serum

Dosage/Administration (Assay Procedure) ＊＊1. Preparation of reagents
Reagent (1): Dissolve NEFA Enzyme Agent 1 withNEFA Buffer Solution 1 before use.
Reagent (2): Disolve NEFA Enzyme Agent 2 withNEFA Buffer Solution 2 before use.
Reagents (1) and (2) are stable for 7 days after

Free fatty acids include oleic acid, palmitic acid,stearic acid, and linoleic acid. Most of these fatty
acids are bound to albumin in the blood.
The serum level of free fatty acids is profoundly
influenced by lipid metabolism, glucose metabolism,and endocrine function, and is increased in patients1reagents 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.
4. Other precautions
Do not use the containers for other purposes.

preparation when stored at 2-10C, under lightshielding circumstance.
2. Assay Procedure
This product is compatible with various types ofautomated analyzer. An example of the assay
procedure is indicated below.
Sample + Reagent (1)7 L150 L
Reagent (2)300 L37C
3 min37C
4.5 min

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

Absorbance I and II: The difference inabsorbance between 660 nm and 546 nm
Calibration material: Anaserum NEFA standard
solution (Manufacture's assigned value)
Reagent blank: Purified water or saline

Assessment of Assay Results ＊＊1. Reference standard range4)
140–850  Eq/L
2. There may be reactions or interfering reactions
with non-target substances. If assay resultsappear to be unreliable, repeat the measurement(if necessary, after dilution) or try anotheranalytical methods.

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 ＊＊Performance
1. Sensitivity
1) Reagent blank: absorbance being equal to orlower than 0.06
2) Sensitivity: The absorbance is 0.24–0.30 per1000  Eq/L of oleic acid.
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)
10–3000  Eq/L
5. Correlation5)
Serum N=106 r=0.998 y=0.96x–20.11
Control method: Approved in vitro diagnostics(enzymatic method)
6. Standard Material
Sodium oleate (Manufacture’s assigned value)

ClinimateNEFAName
NEFA Enzyme
Agent 1(1)NEFA Buffer
Solution 1
NEFA Enzyme
Agent 2(2)NEFA Buffer
Solution 2Package
4×for 50mL
4×50mL
4×for 100mL4×100mL＊＊References ＊1) Itakura H.: Clinical News 2 and 3, 1981.
2) Yamada H, Shimizu M, Tani Y.: Vitamin 54, 189,1980.
3) Maehata E.: Med Tech, 8, 1029, 1980.
4) Kanai M. (supervising editor): Kanai’s manual ofclinical laboratory medicine. 34th ed. 516,Kanehara Shuppan, 2015.
5) In house data, SEKISUI MEDICAL CO., LTD.

＊＊

Precautions for Use or Handling ＊1. Precautions for Handling (to Ensure Safety)
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. 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 suchContact ＊SEKISUI MEDICAL CO., LTD.
international@sekisui.com＊＊

Manufacturer ＊SEKISUI MEDICAL CO., LTD.
1-3, Nihonbashi 2-chome, Chuo-ku, Tokyo, Japan2