Controlled

Title: Standard Operating Procedure for Detecting Lactate

state: Under Page:/17
Dehydrogenase (LDH)
control
No.: XX-SH-001 Version: 1.0 Revision: Revision 0 Effective date:

Approved
Prepared by Reviewed by
by

Preparation Approval
Review Date
Date Date

Standard Operating Procedure for Detecting Lactate

Dehydrogenase (LDH)
1 Purpose
LDH reagent is intended for quantitative determination of Lactate Dehydrogenase in human
serum or plasma on photometric systems. It has important reference value for the diagnosis,
observation of changes in diseases, and treatment effect of relevant clinical diseases.

2 Method& Reaction Principle

2.1 Method
UV-assay according to IFCC (International Federation of Clinical Chemistry
and Laboratory Medicine).
2.2 Reaction Principle
Lactate Dehydrogenase catalyzes the conversion of L-lactate to pyruvate in the presence of
lactate dehydrogenase (LDH). In the process, β-nicotinamide adenine dinucleotide (NAD) is
deoxidized to NADH. This change in absorbance is directly proportional to the activity of LDH
in the sample.

3 Performance characteristics
3.1 Linearity Range and Analytical Specificity

Instrument
BS series chemistry analyzers
Performance Analytical Specificity (mg/dL) (the influence of the
Precision (%) following concentrations of interferents on the results
Chemistry Linearity should not exceed ±10%)
Range
Ascorbic
Within-run Batch-to-batch Hemoglobin Bilirubin Lipemia
acid
4～1000
LDH ≤3.0 ≤5.0 30 / 40 500
U/L
3.2 Precision
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Title: Standard Operating Procedure for Detecting Lactate
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Dehydrogenase (LDH)
control
No.: XX-SH-001 Version: 1.0 Revision: Revision 0 Effective date:

Precision performance using the CLSI Approved Guideline EP5-A2 to assay serum control

appears in the table below. U: U/L

TYPE OF LEVEL II LEVEL III

IMPRECISION Mean SD CV% Mean SD CV%

WITHIN-RUN 1.85 1.32 2.23 1.00

BETWEEN-RUN 1.28 0.91 2.04 0.92

140.71 221.72
BETWEEN-DAY 3.10 2.20 4.50 2.03

WITHIN-DEVICE 3.83 2.72 5.40 2.45

3.3 Comparison

A comparison between Mindray system (Mindray BS series analyzers/Mindray LDH

reagent) (y) and Hitachi/Roche system (Hitachi /Roche LDH, IFCC) (x) using 40 samples gave

following correlation (U/L): y=1.0288x+5.151, R2 = 0.9985.

Details of the comparison experiments are available on request.

4 Specimen collection and preparation

Serum, heparin plasma or EDTA plasma is suitable for sample. Whole blood, hemolysis
and urine are not recommended for use as a sample. Freshly drawn serum is the preferred
specimen.
Use the suitable tubes or collection containers and follow the instruction of the
manufacturer, avoid effect of the materials of the tubes or other collection containers.
Centrifuge samples containing precipitate before performing the assay.
Stability: 4 days at 2－8℃
7 days at 15－25℃
6 weeks at (-15)－(-25)℃

5 Test Requirements
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Title: Standard Operating Procedure for Detecting Lactate
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Dehydrogenase (LDH)
control
No.: XX-SH-001 Version: 1.0 Revision: Revision 0 Effective date:
5.1 Instrument: Mindray BS series biochemical analyzer.
5.2 Reagent: The original reagent is used. Do not mix different batches of reagent kit. The
validity period of the unopened reagent kit stored at 2℃~8℃ is twenty-four months.
Reagent Keep the vial uncapped from light and stable for 30 days on 2℃~8℃. Reagent Do
not freeze. The opened reagent should be used up as soon as possible. The expired reagent
cannot be used.
5.3 Environment requirements (Operating environment requirements of the detection system):
Temperature: 15℃~30℃;
Humidity: 35 %RH~85 %RH, without condensation.
5.4 Safety control: Take necessary precautions when using the monitor. For example, wear
protective suit and gloves. reagent Do not swallow the monitor and avoid contact with the
skin and mucous membrane. reagent and the corresponding calibrators and quality control
products have been tested and confirmed to be negative for HbsAg, HIV and TP antibody of
Treponema pallidum, but for caution, they still need to be handled as infectious dangerous
articles during use. Full attention must be paid to them during operation. Dispose of all
samples and waste in accordance with national and local laws and regulations.

6 Calibration
6.1 Calibrators: Used as matching calibrators:
1) Take out the 2~8℃ storage environment, prepare a proper amount of double distilled
water or deionized water, and balance the water to room temperature;
2) Tap the cap gently to ensure that the lyophilized sample is scattered at the bottom of the
bottle. Carefully open the cap and rubber stopper to avoid loss of the content.
3) Weigh 3 mL of distilled water or deionized water, inject it into the bottle slowly, put on
the rubber stopper and cap, and turn the bottle over for several seconds to keep it away
from light for 30 minutes.
4) Gently invert the vial to ensure that the contents are completely dissolved to avoid foam.
5) After redissolution, select appropriate centrifugal tubes or sample cups for dispensing
and marking.
6) Refer to the Standard Operating Procedure for Mindray Biochemistry Analyzer and use
the reference values in the calibrator reference value table to set the calibration values
and calibrate the analyzer.
Controlled
Title: Standard Operating Procedure for Detecting Lactate
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Dehydrogenase (LDH)
control
No.: XX-SH-001 Version: 1.0 Revision: Revision 0 Effective date:
Unopened calibrators are stored for one year in a dark place under 2~8℃ conditions. The
stability of each test item at different storage temperatures after uncapping and redissolution of
calibrators is shown in the table below. When opening calibrators, tighten the rubber stopper and
cap to avoid contamination and light. Do not freeze or thaw the calibrator repeatedly after
redissolution.

Stabilization Time
Storage temperature
General Items Total bilirubin Direct bilirubin
15～25℃ 8 hours 6 hours 3 hours
2～8℃ 2 days 1 day 8 hours
-25～-15℃ 4 weeks 2 weeks 2 weeks

S1: Physiological saline

S2: Mindray calibrators
6.2 Calibration method: It is recommended to use the Human multi-calibrator from Mindray for
calibration. For details, see《Standard Operating Procedure for Mindray Biochemistry
Analyzer》.
6.3 Calibration period and requirements (including but not limited to the following):
1) When it is used for the first time;
2) Replace the reagent lot number.
3) When replacing key parts;
4) When the indoor QC is out of control.
6.4 Traceability: See the traceability file provided by the manufacturer.

7 Procedure
Prepare the analyzer/reagent preparation → sample ID/data entry → QC → analysis QC
results → sample analysis → analysis results → abnormal result review → test results
review → instrument maintenance and use registration → shutdown of the instrument.
For details about the operations, see 《 Standard Operating Procedure of Mindray
Biochemistry Analyzer》. Analyze and test the collected samples in time. STAT samples shall be
tested first, and the results shall be reported within 30 minutes generally. The report time can be
extended for the samples to be retested. Non-STAT samples should be completed within 2 hours
after blood sampling. When reviewing test results, special attention should be paid to evaluating
their compliance with available patient-related clinical information.
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Title: Standard Operating Procedure for Detecting Lactate
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Dehydrogenase (LDH)
control
No.: XX-SH-001 Version: 1.0 Revision: Revision 0 Effective date:

8 Quality Control
8.1 Control type: The corresponding commercial control provided by the manufacturer.
8.2 QC measure
8.2.1 QC
Run controls before running samples.
1) Take out the 2~8℃ storage environment, prepare a proper amount of double
distilled water or deionized water, and balance the water to room temperature;
2) Tap the cap gently to ensure that the lyophilized sample is scattered at the bottom of
the bottle. Carefully open the cap and rubber stopper to avoid loss of the content.
3) Weigh 5 mL of distilled water or deionized water, inject it into the bottle slowly, cap
the bottle with the rubber stopper, and invert the bottle gently for several seconds.
Keep the bottle away from light for 30 minutes.
4) Gently invert the vial to ensure that the contents are completely dissolved to avoid
foam.
5) After redissolution, select appropriate centrifugal tubes or sample cups for
dispensing and labeling. (The unopened control should be stored in a dark place
under 2~8℃ conditions. The validity period is 18 months.) The stability of the
tested items at different storage temperatures after the open-vial reconstitution of
the control is as follows: When opening the vials of controls, tighten the rubber
stoppers and caps to avoid contamination and light. Do not freeze and thaw the
control after redissolution).
Stabilization Time
Storage temperature Total Bilirubin/Direct Bilirubin
General Items
/Unsaturated iron binding force
15～25℃ 12 hours 8 hours
2～8℃ 5 days 1 day
-25～-15℃ 4 weeks 2 weeks

6) For details about QC test, see the Standard Operating Procedure of Mindray
Biochemistry Analyzer.
The test results of the control shall be within the reference range; otherwise, the test system
shall be checked according to the quality control procedure, such as the control position, validity
period, storage method, calibration process, performance and status of the instrument, etc.
Record the condition of the instrument on the day of quality control.
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Title: Standard Operating Procedure for Detecting Lactate
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Dehydrogenase (LDH)
control
No.: XX-SH-001 Version: 1.0 Revision: Revision 0 Effective date:

9 Principle of Result Calculation Program

Sample Reagent

Reagent color
development
The absorbance principle
changes.
Analysis method

Response

Calibrator
Response
Sample
concentration

10 Calculation
The analyzer calculates the LDH activity of each sample automatically with a specified valid
calibration factor from calibration process. Conversion factor of traditional units (U/L) into
SI-units (μkat/L):
1 U/L = 16.67×10-3 μkat/L, 1 μkat/L= 60 U/L

11 Reference range
Male: ＜248 U/L（＜4.13 μkat/L）;
Female: ＜247 U/L（＜4.12 μkat/L）.

12 Reportable interval of test results

4~ 5000 U/L.
If the value of sample exceeds 1000 U/L, the sample should be diluted with 9g/L NaCl
solution (e.g. 1+ 4) and repeat the assay using this dilution, the result should be multiplied by 5.

13 Warning or Critical Value

For critical values of biochemistry analysis, see 《Standard Operating Procedure for
Reporting Critical Values in the Biochemistry Laboratory》. In case of a critical value, follow this
SOP.

14 Laboratory clinical interpretation

LDH is widely distributed in various tissues or cells of the human body. Among them, the
content of liver, myocardium, kidney, bone marrow muscle, and red blood cells is relatively high
Controlled
Title: Standard Operating Procedure for Detecting Lactate
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Dehydrogenase (LDH)
control
No.: XX-SH-001 Version: 1.0 Revision: Revision 0 Effective date:
and all of them exist in the cytoplasm.
LDH is widely distributed. Therefore, the elevation of serum LDH can be seen in many
clinical conditions, such as myocardial infarction, hepatitis, hemolysis, tumor, kidney, lung, and
muscle. At present, the measurement of serum LDH may be mainly used for the diagnosis of
hematology and tumor-related diseases. Patients with hemolytic anemia (such as cell anemia),
leukemia, or malignant tumor often have significantly elevated serum LDH.

15 References
1) Wroblewski, F & Ledne, J. S, Proc. Soc. Exp Biol Med.96210(1955)
2) Amador. E. Dorfman. L. E. And Acker, W. E. C, Clin. Chem, 9391(1963)
3) Henry, R. J, Chiamorr, N, Golub, O. J. And Berkman, S. Am. J. Clin. Path,
34381(1963)

16 Related Documents
1) Standard Operating Procedure of Mindray Biochemistry Analyzer;
2) Standard Operating Procedure for Laboratory Critical Value Reporting