FRUCTOSE

COD 11794 50 mL

STORE AT 2-8ºC
Reagents for measurement of fructose concentration FRUCTOSE
Only for in vitro use in the clinical laboratory HEXOKINASE/PHOSPHOGLUCOSE ISOMERASE

PRINCIPLE OF THE METHOD Decimals 0 0

No. of replicates 1 1
Fructose in the sample produces, by means of the coupled reactions described below, NADPH Test name in patient report - -
that can be measured by spectrophotometry1,2.
HK PROCEDURE Reading bichromatic bichromatic
D-Fructose + ATP Fructose-6-phosphate + ADP Volumes Sample 12 12
Reagent 1 300 300
PGI Reagent 2 - -
Fructose-6-phosphate Glucose-6-phosphate Washing 1.2 1.2
G6P-DH Predilution factor - -
Glucose-6-phosphate + NADP+ Gluconate-6-phosphate + NADPH + H+ Postdilution factor 2 2
Filters Main 340 340
CONTENTS AND COMPOSITION Reference 670 670
Times Reading 1 300 s 312 s
A. Reagent. 1 x 40 mL. PIPES 70 mmol/L, NADP+ 1.2 mmol/L, hexokinase > 15 U/mL, Reading 2 - -
phosphoglucose isomerase > 10 U/mL, preservatives, pH 7. Reagent 2 - -
B. Reagent. 1 x 10 mL. ATP > 15 mmol/L, glucose 6-phosphate deshydrogenase > 10 U/mL, CALIBRATION Calibration type specific specific
preservatives, pH 9. Calibrator replicates 3 3
S. Fructose standard. 1 x 3 mL. D-fructose 75 mg/dL equivalent to 375 mg/dL (20,8 mmol/L) of Blank replicates 3 3
Calibration curve - -
fructose according to the dilution factor of the sample . Aqueous primary standard.
OPTIONS Blank absorbance limit 0.300 0.300
DANGER: H314: Causes severe skin burns and eye damage. P280: Wear protective Kinetic blank limit - -
gloves/protective clothing/eye protection/face protection. P303+P361+P353: IF ON SKIN (or Linearity limit 1000 1000
hair): Remove/Take off immediately all contaminated clothing. Rinse skin with water/shower.
It is recommended to do a reagent blank every day and a calibration at least every 2 months,
For further warnings and precautions, see the product safety data sheet (SDS). after reagent lot change or as required by quality control procedures.

STORAGE REFERENCE VALUES

Store at 2-8ºC. Seminal plasma: >150 mg/dL = 8.33 mmol/L4
Reagents are stable until the expiry date shown on the label when stored tightly closed and if >2.34 mg/ejaculate = 13 μmol/ejaculate 5
contaminations are prevented during their use. These ranges are given for orientation only; each laboratory should establish its own reference
ranges.
Indications of deterioration:
 Reagents: Presence of particulate material, turbidity, absorbance of the blank higher than QUALITY CONTROL
0.300 at 340 nm (1 cm cuvette). It is recommended to use the Fertility Biochemistry Control (Cod. 18053) to verify the
 Standard: Presence of particulate material, turbidity. performance of the measurement procedure.
Each laboratory should establish its own internal Quality Control scheme and procedures for
REAGENT PREPARATION
corrective action if controls do not recover within the acceptable tolerances.
Working Reagent: Transfer the contents of one Reagent B vial into a Reagent A bottle. Mix
gently. Other volumes can be prepared in the proportion: 4 mL Reagent A + 1 mL Reagent B. METROLOGICAL CHARACTERISTICS
Stable for 3 months at 2-8ºC. The following data were obtained using an A25 analyser. Results are similar with A15. Details
Reagents open and kept in the refrigerated compartment of the analyzer are stable 3 months. on evaluation data are available on request.

ADDITIONAL EQUIPMENT  Linearity limit: 1000 mg/dL = 55 mmol/L. For higher values dilute pretreated sample 1/2 with
distilled water and repeat measurement.
 Thermostatic water bath at 37ºC.
 Detection limit: 4 mg/dL = 0.22 mmol/L.
 Analyzer, spectrophotometer or photometer able to read at 340  20 nm.
 Repeatibility (within run):
SAMPLES Mean Concentration CV n
Allow the fresh semen sample to liquefy at 37ºC for 30 minutes. Centrifuge, in order to separate 122 mg/dL = 6.8 mmol/L 1.4 % 20
the spermatozoa, and decant the seminal plasma3. Fructose in seminal plasma is stable for 6 493 mg/dL = 27.3 mmol/L 0.6 % 20
months at – 20ºC.
 Reproducibility (run to run):
PROCEDURE
Mean Concentration CV n
Sample preparation
122 mg/dL = 6.8 mmol/L 2.0 % 25
The standard do not require pretreatment.
493 mg/dL= 27.3 mmol/L 3.0 % 25
1. Pipette into a test tube:
Seminal plasma 200 L  Trueness: Results obtained with this procedure did not show systematic differences when
Distilled water 800 L compared with a reference procedure. Details of the comparison experiments are available
on request.
2. Shake thoroughly. The sample is stable 8 hours at 15-25ºC, 24 hours at 2-8ºC.
DIAGNOSTIC CHARACTERISTICS
Manual procedure
Fructose is the main sugar in the seminal vesicles and is the carbohydrate energy source of the
1. Bring the Working Reagent to room temperature.
spermatozoa. It is synthesized from blood glucose in the epithelial cells, and its production is
2. Pipette into labelled test tubes: regulated by testosterone.
Blank Standard Sample The concentration levels of fructose depend on the seminal vesicle state and the normal level of
Fructose Standard (S)  50 µL  the hormone. Seminal plasma concentration of fructose reflects the state of the secretory
Sample   50 µL function of the gland. Low concentration levels show ejaculatory duct obstruction or argenenesis
Working Reagent 1.2 mL 1.2 mL 1.2 mL of the vasa differentia and seminal vesicles4,5.

3. Mix thoroughly and incubate the tubes for 10 minutes at room temperature (16-25ºC) or for 5 NOTES
minutes at 37ºC. 1. This reagent may be used in several automatic analysers. Instructions for many of them are
4. Measure the absorbance (A) of the Standard and Sample at 340 nm against the Blank. The available on request.
colour is stable at least 30 minutes.
BIBLIOGRAPHY
5. Calculate fructose concentration using the following general formula:
1. Shimidt, FH. Die enzymatische bestimmung von glucose und fructose nebebeinander. Klin
A Sample x 375 = mg/dL fructose Wschr 1961;39:1244-1247.
A Standard x 20.8 = mmol/L fructose 2. Bergmeyer HU. Methods of enzymatic Analysis, ed Bergmeyer HU, Academic Press, NY,
1974; 3:1196-1201.
Automated procedure (Nota 1)
A25 A15 3. Poirot C, Cherruau. Infertilidad masculina. Aspectos clínicos e investigaciones biológicas.
Acta Bioquim Clin Latinoam 2005;39:225-241.
GENERAL Test name FRUCTOSE FRUCTOSE
Analysis mode mono. end point mono. end point 4. Tietz Textbook of Clinical Chemistry and Molecular Diagnostics, 4th ed. Burtis CA, Ashwood
Sample type SEM SEM ER, Bruns DE. WB Saunders Co, 2005.
Units mg/dL mg/dL 5. WHO laboratory manual for the examination of human semen and sperm-cervical mucus
Reaction type increasing increasing interaction. Cambridge university press, 4th ed, 1999.

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