LDH FS*

DGKC 1970
Diagnostic reagent for quantitative in vitro determination of lactate dehydrogenase (LDH) in serum or
plasma on photometric systems

Order Information Waste Management

Cat. No. Kit size Please refer to local legal requirements.
1 4201 99 10 021 R1 5x 20 mL + R2 1x 25 mL Reagent Preparation
1 4201 99 10 026 R1 5x 80 mL + R2 1x 100 mL
1 4201 99 10 023 R1 1 x 800 mL + R2 1x 200 mL Substrate Start
1 4201 99 10 704 R1 8x 50 mL + R2 8x 12.5 mL The reagents are ready to use.
1 4201 99 10 917 R1 8x 60 mL + R2 8x 15 mL
Sample Start
1 4201 99 90 305 R1 10 x 12 mL + R2 2x 20 mL
Mix 4 parts of R1 + 1 part of R2
Summary [1,2] (eg. 20 mL R1 + 5 mL R2) = monoreagent
Stability: 5 days at 2 – 8 °C
Lactate dehydrogenase (LDH) is an enzyme, consisting of five
8 hours at 15 – 25 °C
different isoenzymes which catalyze the interconversion of
The monoreagent must be protected from light.
L-lactate and pyruvate. LDH is present in the cytoplasm of all
human tissues with higher concentrations in liver, heart and Materials required but not provided
skeletal muscle, and lower values in erythrocytes, pancreas, NaCl solution 9 g/L
kidney and stomach. Increased LDH activities are found in a General laboratory equipment
variety of pathological conditions such as myocardial infarction,
cancer, diseases of liver, blood or muscle. However, because of Specimen
the lack of organ specificity, determination of its isoenzymes or Serum, heparin plasma or EDTA plasma
other enzymes such as alkaline phosphatase or ALAT/ASAT is Stability [4]:
necessary for differential diagnosis. 4 days at 20 – 25 °C
6 weeks at 4 – 8 °C
Method Discard contaminated specimens.
Optimized test according to German Society of Clinical Chemistry
(DGKC) [3]. Assay Procedure
Principle Application sheets for automated systems are available on
request.
< LDH > Lactate + NAD
+ +
Pyruvate + NADH + H
Wavelength 340 nm, Hg 365 nm, Hg 334 nm
Reagents Optical path 1 cm
Temperature 25 °C/30 °C/37 °C
Components and Concentrations Measurement Against air
R1: Phosphate buffer pH 7.5 64 mmol/L Substrate Start
Pyruvate 0.80 mmol/L
R2: Good’s buffer pH 9.6 Temperature 25°C/30°C 37 °C
NADH 1.0 mmol/L Sample/Calibrator 20 µL 10 µL
Reagent 1 1000 µL 1000 µL
Storage Instructions and Reagent Stability Mix, incubate for approx. 1 – 5 min., then add:
The reagents are stable up to the end of the indicated month of Reagent 2 250 µL 250 µL
expiry, if stored at 2 – 8 °C and contamination is avoided. Do not Mix, read absorbance after 1 min. and start stopwatch. Read
freeze the reagents! absorbance again after 1, 2 and 3 min.
Reagent 2 must be protected from light.
Sample Start
Warnings and Precautions
Temperature 25 °C/30 °C 37 °C
1. The reagents contain sodium azide (0.95 g/L) as Sample/Calibrator 20 µL 10 µL
preservative. Do not swallow! Avoid contact with skin and Mono-reagent 1000 µL 1000 µL
mucous membranes. Mix, read absorbance after 1 min. and start stopwatch. Read
2. In very rare cases, samples of patients with gammopathy absorbance again after 1, 2 and 3 min.
might give false results [7].
3. Please refer to the safety data sheets and take the necessary
precautions for the use of laboratory reagents. For diagnostic
purposes, the results should always be assessed with the
patient’s medical history, clinical examinations and other
findings.
4. For professional use only!

LDH FS DGKC – Page 1 * fluid stable

Calculation Precision (at 25 °C)
Intra-assay precision Mean [U/L] SD CV
With factor n = 20 [U/L] [%]
From absorbance readings calculate ∆A/min and multiply by the Sample 1 142 5.50 3.86
corresponding factor from table below: Sample 2 245 4.95 2.01
∆A/min x factor = LDH activity [U/L] Sample 3 497 8.39 1.69
Substrate Start 25 °C/30 °C 37 °C
Inter-assay precision Mean [U/L] SD CV
340 nm 10080 20000
n = 20 [U/L] [%]
334 nm 10275 20390
365 nm 18675 37060 Sample 1 144 3.09 2.13
Sample 2 248 4.53 1.82
Sample Start 25 °C/30 °C 37 °C Sample 3 492 6.23 1.26
340 nm 8095 16030 Method Comparison
334 nm 8250 16345 A comparison of DiaSys LDH FS (y) with a commercially available
365 nm 15000 29705 test (x) using 78 samples gave following results:
With calibrator y = 1.03 x + 2.13 U/L; r= 0.999.

∆A / min Sample Reference Range [6]

LDH [U / L] = x Conc. Calibrator [U / L]
∆A / min Calibrator 25 °C 30 °C 37 °C Unit
Adults: < 240 < 346 < 480 [U/L]
Conversion factor <4 < 5.77 <8 [µkat/L]
LDH [U/L] x 0.0167 = LDH [µkat/L] Each laboratory should check if the reference ranges are
transferable to its own patient population and determine own
Calibrators and Controls reference ranges if necessary.
For the calibration of automated photometric systems, DiaSys
TruCal U calibrator is recommended. This method is traceable to Literature
st
the molar extinction coefficient. DiaSys TruLab N and P controls 1. Thomas L. Clinical laboratory diagnostics. 1 ed. Frankfurt:
should be assayed for internal quality control. Each laboratory TH-Books Verlagsgesellschaft;1998.p.89–94.
should establish corrective action in case of deviations in control 2. Moss DW, Henderson AR. Clinical enzymology In: Burtis CA,
recovery. Ashwood ER, editors. Tietz Textbook of Clinical Chemistry.
rd
3 ed. Philadelphia: W.B Saunders Company;1999.617–721.
Cat. No. Kit size
3. Deutsche Gesellschaft für klinische Chemie. Empfehlungen
TruCal U 5 9100 99 10 063 20 x 3 mL
der deutschen Gesellschaft für Klinische Chemie (DGKC).
5 9100 99 10 064 6 x 3 mL
Standardisierung von Methoden zur Bestimmung von
TruLab N 5 9000 99 10 062 20 x 5 mL
Enzymaktivitäten in biologischen Flüssigkeiten. Z Klin Chem
5 9000 99 10 061 6 x 5 mL
Klin Biochem 1972;10:182-92.
TruLab P 5 9050 99 10 062 20 x 5 mL
4. Guder WG, Zawta B et al. The Quality of Diagnostic Samples.
5 9050 99 10 061 6 x 5 mL st
1 ed. Darmstadt: GIT Verlag; 2001;
p. 36-7.
Performance Characteristics 5. Young DS. Effects of Drugs on Clinical Laboratory Tests. 5th
ed. Volume 1 and 2. Washington, DC: The American
Measuring range Association for Clinical Chemistry Press 2000.
On automated systems the test is suitable for the determination of 6. Fischbach F, Zawta B. Age-dependent reference limits of
LDH activities up to 1200 U/L. several enzymes in plasma at different measuring
In case of a manual procedure, the test is suitable for LDH temperatures. Klin Lab 1992;38:555-61.
activities which correspond to a maximum of ∆A/min of 0.15 at 340 7. Bakker AJ, Mücke M. Gammopathy interference in clinical
and 334 nm or 0.08 at 365 nm. chemistry assays: Mechanism, detection and prevention. Clin
If these values are exceeded the sample should be diluted 1+10 Chem Lab Med 2007; 45(9): 1240–1243.
with NaCl solution (9 g/L) and results multiplied by 11.
Specificity/Interferences Manufacturer
No interference was observed by ascorbic acid up to 30 mg/dL, DiaSys Diagnostic Systems GmbH
IVD Alte Strasse 9 65558 Holzheim Germany
bilirubin up to 40 mg/dL and lipemia up to 2,000 mg/dL
triglycerides. Hemolysis interferes because LDH is released by
erythrocytes. For further information on interfering substances
refer to Young DS [5].
Sensitivity/Limit of Detection
The lower limit of detection is 5 U/L.

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