This package insert follows the Pharmaceuticals, Medical devices and Other Therapeutic Products Act of Japan.
In Vitro Diagnostics
Marketing Notification No. 13A2X00197218099** Revised: January 2017 (10th edition)* Revised: August 2011 (9th edition)

This package insert must be read carefully prior to use.

Lactate dehydrogenase assay kit(Classification No.: 38504000)

2. Features
1) The final concentration of the active
ingredient is identical to that mentioned in theJSCC-recommended method.
2) Applicable to various automated analyzers.

Pureauto S LD
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.

Procedural Precautions ＊＊1. Properties of Samples and Sampling Methods
1) Samples
Serum and plasma (heparin plasma andcitrated plasma) may be used.
2) Storage of samples
If the isolated serum or plasma sample cannot
be tested on the same day, specimens should
be stored as follows.4)
Room temperature: for tests within 1 week
Please note that activity of samples may
decrease when refrigerated or frozen.
2. Interfering substances
Assay results are not affected by free bilirubin (upto 40 mg/dL), conjugated bilirubin (up to 40mg/dL), hemoglobin (up to 500 mg/dL), formazinturbidity (up to 5600 FTU), or ascorbic acid (upto 50 mg/dL). Because LD is contained in blood
cells, hemolyzed samples show artifactual
elevation of the LD level.
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.

Description (Kit Components) ＊＊Component: Ingredients
LD Substrate Solution 1:
Lithium L-lactate
Diethanolamine buffer
LD Coenzyme Solution 2:
Nicotinamide adenine
dinucleotide (oxidized form)

Dosage/Administration (Assay Procedure) ＊＊1. Preparation of reagents
Reagent (1): LD Substrate Solution 1 is ready touse.
Reagent (2): LD Coenzyme 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.

Intended Use
Measurement of lactate dehydrogenase in serumor plasma
Lactate dehydrogenase catalyzes hydrogen transfer
reactions between lactic acid and pyruvic acid. It is
widely distributed in all tissues of the body.
Since the blood level of lactate dehydrogenase isincreased in various diseases of the heart, liver, andkidneys, as well as various solid tumors, leukemia,
and pernicious anemia, it plays an important role inthe diagnosis and follow-up of these diseases.

SampleReagent (1)+10 L260 L
Reagent (2)130 L

Assay Principle
1. Assay Principle
In samples, lactate dehydrogenase (LD) catalyzes
conversion of lactic acid to pyruvic acid in adiethanolamine buffer. The reaction also converts
NAD to NADH, and its absorbance at 340 nm
increases. LD activity is determined bymeasuring the velocity of NADH production.
LDLactic acid + NAD
Pyruvic acid + NADH

37C
5 min37C
60-180 sec

Measurement(Absorbance※)
Calculation ofactivity※

1

Absorbance: The difference in absorbance
between 405 nm and 340 nm
Calibration material: Enzyme Calibrator Plus
“Daiichi” (Manufacturer's assigned value)
Reagent blank: Purified water or saline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.
5) Sodium azide is added as an antiseptic agent in
the LD Coenzyme Solution 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
when disposing.
4. Other precautions
Do not use the containers for other purposes.

Assessment of Assay Results ＊＊1. Reference standard range 5)124‐222 U/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.
Performance ＊1. Sensitivity
1) Reagent blank: change in absorbance being
equal to or lower than 0.005/min
2) Sensitivity: The change in absorbance is0.029–0.035/min per 200 U/L of LD 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 7600Ds
automated analyzer)
3–900 U/L
5. Correlation 7)
1) Serum N=70 r=0.999 y=1.00x＋1.1
Control method: using an already approved invitro diagnostic product with method in linewith the JSCC Recommendation for Enzyme
Activities.
2) Plasma N=95 r=0.999 y=1.00x＋4.6
Control method: using an already approved invitro diagnostic product with method in linewith the JSCC Recommendation for EnzymeActivities.
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
LD Substrate(1)
4 × 100 mL
Solution 1
Pureauto S
LDLD Coenzyme(2)
4 × 100 mL
Solution 2
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 LD Coenzyme Solution 2. Therefore, if thereagent comes in accidentally contact with eyes,mouth or skin, rinse immediately with ample
water as first aid, and consult the doctor ifrequired.
2. Precautions for use
1) This product should be stored as directed, avoidfreezing. Freezing can cause deterioration ofthe reagents, leading to inaccurate results.
Therefore, do not use the product if it has beenpreviously 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.

References ＊＊1) Japan Society of Clinical Chemistry: Jpn J Clin
Chem, 19, 228, 1990.
2) Japan Society of Clinical Chemistry: Jpn J Clin
Chem, 19, 232, 1990.
3) Osawa S.: Med. Tech, 20, 1022, 1992.
4) Sasaki M. et al.: Sampling of constituents of thehuman body, 217, Kodansha, 1972.
5) Kanai M. (supervising editor): Kanai’s manual ofclinical laboratory medicine. 34th ed. 581,
Kanehara Shuppan, 2015.
6) Japan Society of Clinical Chemistry: Jpn J Clin
Chem, 25, 135, 1996.
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