1141010I Rev. 02

LDH BR
REF 1141010
2 x 50 mL LDH BR
SFBC
CONTENTS UV enzymatic method
R1. Reagent 2 x 40 mL
R2. Reagent 1 x 20 mL KINETIC
For in vitro diagnostic use only
PRINCIPLE INTERFERENCES
Lactate dehydrogenase (LD/LDH) catalyzes the reduction of  Lipemia (intralipid  20 g/L) does not interfere.
pyruvate to lactate (P-L) in the presence of reduced nicotinamide
 Bilirubin ( 40 mg/dL) does not interfere.
adenine dinucleotide (NADH) at pH 7.5.
The reaction is monitored kinetically at 340 nm by the rate of  Hemoglobin ( 4 g/L) may affect the results.
decrease in absorbance resulting from the oxidation of NADH to  Other drugs and substances may interfere3.
NAD+ proportional to the activity of LD present in the sample.
LD/LDH MATERIALS REQUIRED

Pyruvate + NADH + H+ L-Lactate + NAD+
pH 7.5
 Photometer or spectrophotometer with a thermostatted cell
compartment set at 30/37ºC, capable of reading at 340 nm.
This test has been formulated according the standarized method  Stopwatch, strip-chart recorder or printer.
described by SFBC.1
 Cuvettes with 1-cm pathlength.
 Pipettes to measure reagent and samples.
REAGENT COMPOSITION
R1 LDH substrate. TRIS buffer 100 mmol/L pH 7.5, pyruvate PROCEDURE

2.75 mmol/L, sodium chloride 222 mmol/L.
1. Preincubate working reagent, samples and controls to reaction
R2 LDH coenzyme. NADH 1.55 mmol/L. temperature 30/37ºC.
2. Set the photometer to 0 absorbance with distilled water.
3. Pipette into a cuvette:
STORAGE AND STABILITY
Reaction temperature 30/37ºC
Store at 2-8ºC.
All the kit compounds are stable until the expiry date stated on the Working reagent 1.0 mL
label. Do not use reagents over the expiration date. Sample or control 20 L
Store the vials tightly closed, protected from light and prevented
contaminations during the use.
Discard If appear signs of deterioration: 4. Mix gently by inversion. Insert cuvette into the cell holder and
- Presence of particles and turbidity. start stopwatch.
- Blank absorbance (A) at 340 nm < 1.000 in 1cm cuvette. 5. Incubate for 30 seconds and record initial absorbance reading.
6. Repeat the absorbance readings exactly after 1, 2 and 3
minutes.
REAGENT PREPARATION 7. Calculate the difference between absorbances.
8. Calculate the mean of the results to obtain the average change
in absorbance per minute (A/min).
Working reagent. Mix 4 mL of R1 + 1 mL of R2. Stable for 2 months
at 2-8ºC.
Protected from light.
CALCULATIONS
SAMPLES
U/L = A/min x 8095
Serum free of hemolysis separated from the cells as soon as Samples with A/min exceeding 0.150 at 340 nm should be diluted
possible after collection. The use of heparin and citrate as 1:10 with saline and assayed again. Multiply the results by 10.
anticoagulants have been reported to falsely elevate LD activity. If results are to be expressed as SI units apply:
Freezing results in loss of activity of the enzyme. U/L x 0.01667 = kat/L
QUALITY SYSTEM CERTIFIED LINEAR CHEMICALS, S.L.U. Joaquim Costa 18 2ª planta. 08390 Montgat (Barcelona) SPAIN
ISO 9001 ISO 13485 Telf. (+34) 934 694 990; E-mail: [email protected] ; website: www.linear.es NIF-VAT:B60485687
REFERENCE VALUES4 NOTES
Serum 1. This method may be used with different instruments. Any
application to an instrument should be validated to demonstrate
that results meets the performance characteristics of the
Temperature 37ºC 30ºC
method. It is recommended to validate periodically the
207 - 414 U/L 140 - 280 U/L instrument. Contact to the distributor for any question on the
Adults application method.
(3.40 - 6.80 Kat/L) (2.30 - 4.70 Kat/L)
2. Clinical diagnosis should not be made on findings of a single test
It is recommended that each laboratory establishes its own result, but should integrate both clinical and laboratory data.
reference range.
REFERENCES
QUALITY CONTROL
To ensure adequate quality control (QC), each run should include a 1. Comission Enzymologie de la Societé Française de Biologie
set of controls (normal and abnormal) with assayed values handled Clinique. Ann. Biol. Clin. 40: 123-128 (1982).
as unknowns. 2. Sociedad Española de Química Clínica, Comité Científico,
Comisión de Enzimas. Método recomendado para la
REF 1980005 HUMAN MULTISERA NORMAL determinación en rutina de la concentración catalítica de
Borderline level of LDH. Assayed. lactato deshidrogenada en suero sanguíneo humano. Quim
Clin 1988; 8:57-61.
REF 1985005 HUMAN MULTISERA ABNORMAL 3. Young DS. Effects of drugs on clinical laboratory tests, 4th ed.
Elevated level of LDH. Assayed. AACC Press, 1995.
4. Tietz. N.W. Clinical Guide to Laboratory Tests, 3rd Edition.
If the values are found outside of the defined range, check the W.B. Saunders Co. Philadelphia, PA. (1995).
instrument, reagents and procedure. 5. Friedman and Young. Effects of disease on clinical laboratory
Each laboratory should establish its own Quality Control scheme tests, 5th ed. AACC Press, 2000.
and corrective actions if controls do not meet the acceptable
tolerances.
CLINICAL SIGNIFICANCE2,5
The enzyme activity found in circulation is a mixture of five

isoenzymes. Each organ has a characteristic isoenzyme profile.
Leakage of these isoenzymes from a diseased organ results in the
elevation of total serum LD.
Increased levels become evident 8-12 hours after myocardial
infarction reaching a maximum 4-5 days later. Elevated values in
serum are also encountered in cases of pulmonary embolism and in
about one third of patients with renal disease, specially those with
pyelonephritis or tubular necrosis. In toxic hepatitis with jaundice,
Hodking´s disease and abdominal and lung cancers elevations are
specially high.
Moderate increases are also observed in cases of liver disease,
megaloblastic and pernicious anemia and progressive muscular
dystrophy.
Decreases are not important clinically.
ANALYTICAL PERFORMANCE
- Detection Limit : 15.73 U/L

- Linearity : Up to 1500 U/L
- Precision:
U/L Within-run Between-run

Mean 495 879 495 879
SD 4.32 7.34 14 13.5
CV% 0.87 0.83 2.83 1.54
N 10 10 10 10
- Sensitivity : 0.152 mA / U/L LDH

- Correlation: This assay (y) was compared with a similar commercial
method (x). The results were:
N =52 r = 0.99 y = 1.031x + 3.147
The analytical performances have been generated using on automatic

instrument. Results may vary depending on the instrument. B1141-2/0901
R1.ing
QUALITY SYSTEM CERTIFIED LINEAR CHEMICALS, S.L.U. Joaquim Costa 18 2ª planta. 08390 Montgat (Barcelona) SPAIN
ISO 9001 ISO 13485 Telf. (+34) 934 694 990; E-mail: [email protected] ; website: www.linear.es NIF-VAT:B60485687

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1141010I Rev. 02

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LDH BR

LD/LDH MATERIALS REQUIRED

R1 LDH substrate. TRIS buffer 100 mmol/L pH 7.5, pyruvate PROCEDURE

The enzyme activity found in circulation is a mixture of five

- Detection Limit : 15.73 U/L

U/L Within-run Between-run

- Sensitivity : 0.152 mA / U/L LDH

The analytical performances have been generated using on automatic

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