This package insert follows the Pharmaceuticals, Medical devices and Other Therapeutic Products Act of Japan.
In Vitro Diagnostics
Marketing Notification No. 13A2X00197218084**Revised: January 2017 (8th edition)*Revised: August 2011 (7th edition)

This package insert must be read carefully prior to use.
Alkaline phosphatase assay kit for blood(Classification No.: 33165001)

matches with the final concentration stipulated
in the JSCC Recommendation for EnzymeActivities.
2) Applicable to various automated analyzers.

Autosera S ALPProcedural Precautions ＊＊1. Properties of Samples and Sampling Methods
1) Samples
Serum and plasma (heparin plasma andcitrated plasma) may be used.
2) Storage of samples
The isolated serum (plasma) should be testedon the same day.
If samples cannot be measured on the sameday, storage should be performed at -20C orlower.5)
Bring samples to room temperature (15–30C)
before use.
2. Interfering substances
Assay results are not affected by free bilirubin (upto 40 mg/dL), conjugated bilirubin (up to 40
mg/dL), hemoglobin (up to 500 mg/dL), formazinturbidity (up to 5600 FTU), or ascorbic acid (upto 50 mg/dL).
3. Others
1) Always use Enzyme Calibrator Plus “Daiichi”
for calibration.
2) Precautions for assay range
If the activity of sample exceeds assay range,
dilute the sample with saline and repeat themeasurement.

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 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.
Description (Kit Components) ＊＊Component
Ingredients
ALP Reagent 1: Ethylaminoethanol Buffer Solution
Magnesium chloride
ALPReagent 2: Disodium 4-nitrophenyl phosphate

Dosage/Administration (Assay Procedure) ＊＊1. Preparation of reagents
Reagent (1): ALP Reagent 1 is ready to use.
Reagent (2): ALP 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.

Intended Use
Measurement of alkaline phosphatase in serum orplasma
Alkaline phosphatase (ALP) is an enzyme thathydrolyzes phosphate compounds at the optimal pH(alkaline). ALP activity is high in organs/tissues suchas the kidneys (proximal uriniferous tubule), small
intestine (mucosa), osteoblasts, placenta, liver, andmammary gland. ALP activity is increased in bonedisease, hepatobiliary disease, pregnancy, andmalignancies.

Sample + Reagent (1)4 μL
260 μL

37C
5 min
37C

Measurement
Reagent (2)※130 μL 96–300 sec (Absorbance )Calculation ofactivity

Assay Principle
1. Assay Principle
4-Nitrophenol (yellow color) is released when
ALP in samples acts on 4-nitrophenyl phosphateas the substrate in Ethylaminoethanol (EAE)
Buffer Solution. ALP activity is determined bymeasuring the velocity of 4-nitrophenol
production.
ALP
4-Nitrophenyl phosphate ─▶2+4-Nitrophenol
Mg(Yellow color) + Phosphoric acid
2. Features1) Final concentration of the principal ingredient※

Absorbance: The difference in absorbance
between 505 nm and 405 nm.
Calibration material: Enzyme Calibrator Plus
“Daiichi” (manufacture's assigned value)
Reagent blank: Purified water or saline

Assessment of Assay Results ＊＊1. Reference standard range6)
106–322 U/L
2. There may be reactions or interfering reactions
with non-target substances. If assay results appear
to be unreliable, repeat the measurement (if1necessary, after dilution) or try another analyticalmethods.

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) Sodium azide has been added as an antiseptic
agent in the ALP Reagent 2. It can react withlead or copper pipes to produce the highly
explosive metal azide. Therefore, the reagent
should be flushed with large amounts of water
during disposal.
4. Other precautions
Do not use the containers for other purposes.

Performance ＊1. Sensitivity
1) Reagent blank: change in absorbance being
equal to or lower than 0.002/min
2) Sensitivity: The change of absorbance is0.010–0.030/min per 500 U/L of ALP activity.
2. Accuracy: 90–110 % of the expected assay value
3. Within-run Reproducibility:
Coefficient of variation ≤ 5 %(Test methods used for 1.–3. are in-housemethods.)
4. Measurement Range7): (On Hitachi 7170S
automated analyzer)
5–2000 U /L
5. Correlation7)
1) Serum N=50
r=0.999
y=1.05x–3.1
Control method: using an already approved invitro diagnostic product with method in line with
the JSCC Recommendation for Enzyme Activities
2) Plasma N=70
r=0.999
y=0.99x–0.6
Control method: using an already approved invitro diagnostic product with method in line with
the JSCC Recommendation for Enzyme Activities
6. Standard Material
Enzyme Calibrator Plus, the calibration material
used for this product, is in line with the JapaneseStandard for Certified Enzyme ReferenceMaterials.

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
ALP Reagent 1
4 × 85 mL(1)
ALP Reagent 1
4 × 50 mL
Autosera S
ALP
ALP Reagent 2
4 × 45 mL(2)
ALP Reagent 2
4 × 50 mL
Constituent reagents are available in otherconfigurations. For further details please contact
SEKISUI MEDICAL CO., LTD.

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) Sodium azide is added as an antiseptic agent in
the ALP Reagent 2. Therefore, if the reagent
comes in accidentally contact with eyes, mouthor skin, rinse immediately with ample water as
first aid, and consult the doctor if required.
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 121 ºC 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 %.

References ＊＊1) Japan Society of Clinical Chemistry: Jpn J Clin
Chem, 19, 209, 1990.
2) Japan Society of Clinical Chemistry: Jpn J Clin
Chem, 19, 213, 1990.
3) Osawa S.: Med Tech, 20, 1022, 1992.
4) Sasaki M. et al.: Sampling of constituents of thehuman body, 170, Kodansha, 1972.
5) Nakahara M. et al.: Jpn J Med Tech, 45, 1106,
1996.
6) Kanai M. (supervising editor): Kanai’s manual ofclinical laboratory medicine. 34th ed. 558,Kanehara Shuppan, 2015.
7) 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, Japan2