For in vitro diagnostic use only. Do not pipette by mouth.
MANUAL Exercise the normal precautions required for handling laboratory RX MONZA reagents.
INTENDED USE Solution R1a contains picric acid which is poisonous.
For the quantitative in vitro determination of creatinine in serum, Solution R1b contains sodium hydroxide which is caustic. plasma or urine. This product is suitable for manual use and on the Rx Monza analyser. Solution R1b is classified as hazardous according to Regulation (EC) No. 1272/2008 (CLP). Cat. No. CR 510 CAL. Standard 1 x 5.5 ml 200 ml R1a. Picric Acid 1 x 100 ml R1b. Sodium Hydroxide 1 x 100 ml
GTIN: 05055273201901
CR 524 CAL. Standard 1 x 30 ml
6 x 500 ml R1a. Picric Acid 3 x 500 ml R1b. Sodium Hydroxide 3 x 500 ml DANGER
GTIN: 05055273201918 Causes severe skin burns and eye damage.
Creatinine is derived from creatine and creatine phosphate in protection. muscle tissue and may be defined as a nitrogenous waste IF ON SKIN (or hair): Take off immediately all contaminated product. Creatinine is not reutilised but is excreted from the clothing. Rinse skin with water/shower. body in the urine via the kidney. It is produced and excreted at a IF IN EYES: Rinse cautiously with water for several minutes. constant rate which is proportional to the body muscle mass. As Remove contact lenses, if present and easy to do. Continue a consequence of the way in which creatinine is excreted by the rinsing. kidney, creatinine measurement is used almost exclusively in the Store locked up. assessment of kidney function. Creatinine is regarded as the most useful endogenous marker in the diagnosis and treatment Safety data sheets available on request. of kidney disease. Creatinine is measured primarily to assess kidney function and Please dispose of all Biological and Chemical materials according has certain advantages over the measurement of urea. The to local guidelines. plasma level of creatinine is relatively independent of protein ingestion, water intake, rate of urine production and exercise. The reagents must be used only for the purpose Since its rate of production is constant, elevation of plasma intended by suitably qualified laboratory personnel, creatinine is indicative of under-excretion, suggesting kidney under appropriate laboratory conditions. impairment. Depressed levels of plasma creatinine are rare and not clinically significant. STABILITY AND PREPARATION OF REAGENTS CAL. Standard COLORIMETRIC METHOD(1) Supplied ready to use. Stable to expiry date when stored at +2 to +25°C. PRINCIPLE Creatinine in alkaline solution reacts with picric acid to form a R1a. Picric Acid coloured complex. The amount of the complex formed is Supplied ready to use. Stable to expiry date when stored directly proportional to the creatinine concentration. at +15 to +25°C.
SAMPLE COLLECTION AND PREPARATION R1b. Sodium Hydroxide
Serum, Heparinized or EDTA plasma. Stable for 7 days at +2 to Supplied ready to use. Stable to expiry date when stored + 8°C. at +15 to +25°C. Urine collected without additives. Urine samples should be diluted 1 + 49 with distilled water. Stable for 4 days at +2 to STABILITY AND PREPARATION OF WORKING +8°C. REAGENT Mix equal volumes of Solutions R1a + R1b. Stable for 3 days at REAGENT COMPOSITION +15 to +25°C.
Contents Initial Concentrations of Solutions MATERIALS PROVIDED
Standard CAL. Standard See lot specific insert Picric Acid R1a. Picric Acid 35 mmol/l Sodium Hydroxide R1b. Sodium Hydroxide 0.32 mol/l
PAGE 1 OF 4 MANUAL/ RX MONZA CR 510
MATERIALS REQUIRED BUT NOT PROVIDED Concentration of creatinine in urine.
Pipetting devices for the delivery of 50 µl, 100 µl, 200 µl, 1 ml Asample and 2 ml. x Standard = mmol/l Timing device and water bath or heating block to maintain Astandard conc.(µmol/l) x 0.05 temperature at +25, +30 or +37°C. Spectrophotometer with wavelength capability of 490 to 510 nm. Asample Randox Assayed Multisera Level 2 (Cat. No. HN 1530) and Standard x conc (mg/dl) x 50 = = mg/dl Level 3 (Cat. No. HE 1532). Astandard Randox Calibration Serum Level 3 (Cat. No. CAL 2351). mg creatinine/dl urine Creatinine x ml urine 24 hrs PROCEDURE NOTES Clearance = [ml/min] Reaction rate and absorbance of the reaction product are very mg creatinine/dl serum x 1440 sensitive to temperature. The specified temperature must therefore be maintained. STANDARDISATION Randox Calibration Serum Level 3 is traceable to creatinine PROCEDURE reference materials NIST 909b and NIST 967. Using fresh ddH2O perform a new Gain Calibration in cuvette mode. Select CREA in the Run Test screen and carry out a water QUALITY CONTROL blank as instructed. Randox Assayed Multisera, Level 2 and Level 3 are recommended for daily quality control. Two levels of controls should be assayed Pipette into a cuvette: at least once a day. Values obtained should fall within a specified range. If these values fall outside the range and repetition excludes Reagent Blank S0 Standard S1 Sample error, the following steps should be taken: 1. Check instrument settings and light source. ddH2O 50 µl - - 2. Check cleanliness of all equipment in use. Standard - 50 µl - 3. Check water, contaminants i.e. bacterial growth may contribute Sample - - 50 µl to inaccurate results. Working Reagent 500 µl 500 µl 500 µl 4. Check reaction temperature. 5. Check expiry date of kit and contents. Mix, insert into the Rx Monza flowcell holder and press Read. 6. Contact Randox Laboratories Technical Services, Northern Ireland +44 (0) 28 94451070. FOR MANUAL USE INTERFERENCE Wavelength: 492 (490-510 nm) Haemolysis interferes with the test. Do not use lipaemic sera. Cuvette: 1 cm light path The method is subject to interferences from high levels of Temperature: +25/+30/+37°C reducing substances. Boiling urine before testing may help reduce Measurement: against air this interference. Pipette into cuvette: Physiological changes in serum or plasma analyte concentrations can be caused by a number of substances. Comprehensive Standard Sample discussion of possible interfering substances, their serum or Macro Semi Macro Semi plasma concentrations, and their possible physiological Micro Micro involvements is beyond the scope of this document. The listed reference contains specific details on known potential interfering Working reagent 2.0 ml 1.0 ml 2.0 ml 1.0 ml substances (4). The user must remain vigilant to the possible Standard solution 0.2 ml 0.1 ml - - effect on results of unknown interferences from medications or Sample - - 0.2 ml 0.1 ml endogenous substances. All patient results must be evaluated in light of the total clinical status of the patient. Mix and after 30 seconds read the absorbance A1 of the standard and sample. Exactly 2 minutes later, read absorbance A2 of NORMAL VALUES(2) standard and sample. Serum: Men 53 - 97 µmol/l (0.6 - 1.1 mg/dl) Women 44 - 80 µmol/l (0.5 - 0.9 mg/dl) CALIBRATION FOR RX MONZA Urine: 8.84 - 13.3 mmol/24 hrs Recommended on change of reagent lot or as indicated by quality 1 - 1.5 g/24 hrs control procedures, using supplied CAL Standard in kit or Randox Calibration Serum Level 3. It is recommended that each laboratory establish its own reference range to reflect the age, sex, diet and geographical MANUAL CALCULATION location of the population. A2 - A1 = A sample or A standard SPECIFIC PERFORMANCE CHARACTERISTICS Concentration of creatinine in serum or plasma. The following performance data were obtained using an Rx Asample Monza analyser running at +37°C. x Standard = µmol/l Astandard conc.(µmol/l)
Asample Standard x conc (mg/dl) = mg/dl Astandard
PAGE 2 OF 4 MANUAL/ RX MONZA CR 510
SERUM
LINEARITY If the concentration exceeds 2168 mol/l (24.5 mg/dl) in serum, dilute serum, 1+4 with 0.9% (w/v) NaCl and repeat the assay. Multiply the result by 5.
SENSITIVITY The minimum detectable concentration of Creatinine with an acceptable level of precision was determined as 14.0 mol/l (0.158 mg/dl).
PRECISION
Intra Assay Level 2 Level 3 Mean (mg/dl) 1.46 4.19 SD 0.040 0.068 CV (%) 2.75 1.63 n 20 20
Inter Assay Level 2 Level 3 Mean (mg/dl) 1.46 4.19 SD 0.042 0.122 CV (%) 2.86 2.91 n 20 20
CORRELATION This method (Y) was compared with another commercially available method (X) and the following linear regression equation obtained:
Y = 1.098 X - 0.27 and a correlation coefficient of r = 0.9995
47 patient samples were analysed spanning the range 0.24 to
15.3 mg/dl.
URINE
LINEARITY If the concentration exceeds 85309 µmol/l (963 mg/dl) in urine, dilute urine 1+4 with distilled water and repeat the assay. Multiply the result by 5.
SENSITIVITY The minimum detectable concentration of Creatinine with an acceptable level of precision was determined as 2221 µmol/l (25.1 mg/dl).
REFERENCES 1. Bartels, H., Bohmer, M., (1972) Clin. Chem. Acta 37: 193. 2. Schirmeister, J., H. Willmann, and H. Kiefer. (1964). Dtsch. Med. Wschr. 89: 1018. 3. Young DS. Effects of Drugs on Clinical Laboratory Tests. 5th ed. Washington, DC: AACC Press; 2000.
The presence of a vertical bar in the margin indicates a technical
