Intended use . Colorimetric system for creatinine determination in sodium tetraborate (12.7 mmol/L) and surfactant.

rfactant. The reagent may

blood (serum, plasma) and urine by end point reaction. precipitate in low temperatures at 15 ºC. In this case, warm up at 37 ºC
and mix until it dissolves completely.
Professional use.
3. - Standard 4.0 mg/dL - Store at 15 - 25 ºC.
[For in vitro diagnostic use.] Contains 4.0 mg/dL creatinine, solubilizer and 0.1% preservative.
Reagent label bears expiration date. In order to avoid evaporation of the
Test principle . Creatinine and other components of serum react to Standard, keep the bottle tightly closed.
alkaline picrate in alkaline solution yielding a red complex what is
photometrically measured. 4. - Acid solution - Store at 15 - 25 ºC.
The addition of an acid decrease the pH to 5.0 yielding the decomposition Acetic acid (≤ 15 mol/L).
of the creatinine picrate, the chromogen derived color remained unaltered
what is also a photometric measurement. The difference of the two Precautions and warnings
measurements yields the true value of creatinine.
For in vitro diagnostic use.
Summary . Creatinine applies an end point procedure in order to
Disposal of all waste material should be in accordance with local
improve the method specificity and minimize the susceptibility to
1-3,5 guidelines.
interferences .
The usual security cares should be applied on the reagent handling.
The measurement procedure is calibrated with the NIST SRM 914 and
renders the results traceable to the IDMS (isotropic dilution, mass The reagents Buffer and Acid solution are corrosive. Avoid ingestion. In
spectrometry) definitive method, which complies the National Kidney case of eyes, skin and mucosa contact, immediately flush then with plenty
Disease Education Program (NKDEP) recommendations for of water and get medical assistance.
4
standardization of serum creatinine measurement .
The Buffer may result in ulcerations when ingested. In case of ingestion,
All the direct methods that apply the Jaffe reaction are susceptible to a immediately offer a lot of water with lemon juice or vinegar. Do not induce
constant systematic error, due to the plasmatic proteins ant other vomiting and get medical assistance.
chromogens interference. In order to minimize this error and increase the
Creatinine results accuracy, Labtest recommends the use of the In case of ingestion of Picric Acid, offer 4 glasses of water and if the
8,11
correction index that must be applied whatever are the found results . individual is conscious, induce vomiting and get medical assistance.

Several studies have demonstrated that is possible minimize significantly Storage and stability . Unopened reagents, when stored at
the interferences caused by icteric and lipemic samples in creatinine indicated temperature, are stable up to expiration date shown on the label.
3
measurement . The procedure with acidification step minimizes the
bilirubin negative interference, while the desproteinization procedure Deterioration . Microbial or chemical contamination may decrease
removes interference of lipemic samples equivalent to a triglycerides reagents stability.
11
value of 250 up to 1800 mg/dL .
Specimen collection and preparation
The measurement procedure is applied in automated and semi-
automated systems able to perform accurate measure of absorbance at Use serum or plasma (heparin, EDTA, fluoret, oxalate and citrate). The
510 nm. analyte is reportedly stable for about 7 days at 2 - 8 ºC.

Methodology . Labtest. Glistab anticoagulant (Labtest Ref.: 29) allows the collection of only one
blood sample for the measurements of creatinine, glucose, and urea.
Reagents Urine of 24 hours must be stored at 2 - 8 ºC during the period of collection
until it is measured.
1. - Picric acid - Store at 15 - 25 ºC.
Reagent label bears expiration date. Picric acid ≤ 50 mmol/L. No known test method can offer complete assurance that human blood
samples will not transmit infectious diseases. Therefore, all blood
2. - Buffer - Store at 15 - 25 ºC. derivatives should be considered potentially infectious.
Reagent label bears expiration date. Sodium hydroxide (208 mmol/L),

01 English - Ref.: 35
 Acid solution (Nº. 4) 0.1 mL 0.1 mL
Interference
Proteins present in the samples yield a positive interference introducing a
constant systematic error. This error may be minimized applying a Mix and let at room temperature for 5 minutes. Measure the absorbance of
correction index. the Unknown against Blank at 510 nm or green filter (500 - 540 nm).
Since the urine has no proteins that may interfere, the correction index is Absorbance will be A2.
not applied to the calculation of concentration in urine samples. See
application of correction index on Calculations. Calculations
Creatinine determination in urine samples may be affected by the action of A1 - A2
high amount of reducers substances present in cases of ketoacidosis. Creatinine (not corrected) = x 4 mg/dL
Boiling the urine sample for one minute eliminates partially these Astandard
substances interference. The remaining interference is excluded in the
4
procedure with acidification step. According with NKDEP recommendations, the results should be reported
with two places of decimals in order to avoid systematic errors due to
For therapeutic control it is recommended to collect the sample at the making the results round, which may reach ± 6%.
same time due circadian variations.
The aspirin in anti-inflammatory doses increases the creatinine value in Due the great reproductive results of the assays system, it is possible to
blood sample. use the factor method:
Physical exercises increase the creatinine values.
Creatinine values are lower in individuals who have vegetarian diet. Calibration factor = 4 /Astandard

Bilirubin up to 5 mg/dL, hemoglobin up to 180 mg/dL and triglycerides up Creatinine (not corrected) = (A1 - A2) x Factor (mg/dL)
to 250 mg/dL do not interfere significantly.
Urinary creatinine
Bilirubin values over 5 mg/dL interfere negatively in the reaction.

Triglycerides values over 250 mg/dL provide false increased results. Urinary Creatinine
Urinary
(mg/24 hours) = x UrineVolume (mL/24h)
Creatinine
Materials required not provided 100

1. A constant temperature water bath (37 ºC). mg/kg weight = mg/24 hours divided by body weight
2. Photometer capable of measuring absorbance at 500 - 540 nm.
3. Pipettes to measure reagents and samples. Applying the correction index . Plasmatic proteins interference
5
4. Timer. that occurs in Jaffe reaction, introduce a constant error in the
measurement which is minimized by the correction index (0.25 mg/dL).
Manual and direct assay procedure The obtained results with the calibration and the correction are
4
traceable to IDMS method and comply with the NKDEP
See notes 1, 2 and 3. recommendations.

Urine . Dilute the urine 1:25 with distilled water (0.2 mL of urine + Creatinine = Creatinine - Correction index
4.8 mL of distilled water) and multiply the result by the dilution factor (25). (corrected) (not corrected) (0.25 mg/dL)
The water must have resistivity ³1 megaohm, or conductivity
£1 microsiems and silicates concentration must be <0.1mg/L. Procedure with deproteinization

Set up three tubes and proceed as follows: This must be applied to icteric and turbid samples.

Mix 0.5 mL of serum or plasma to 1.0 mL of Picric Acid (n° 1),

Blank Unknown Standard homogenize and centrifuge 10 minutes.
Buffer (Nº. 2) 2.0 mL 2.0 mL 2.0 mL
Sample 0.25 mL Set up three tubes and proceed as follow
Distilled or deionized water 0.25 mL
Standard (Nº. 3) 0.25 mL Blank Unknown Standard
Picric acid (Nº. 1) 0.5 mL 0.5 mL 0.5 mL Buffer (Nº. 2) 2.0 mL 2.0 mL 2.0 mL
Supernatant 0.75 mL
Mix and incubate in a water bath at 37 ºC during 10 minutes. Measure the Distilled or deionized water 0.25 mL
absorbance of the Unknown and Standard against Blank at 510 nm or Standard (Nº. 3) 0.25 mL
green filter (500 - 540 nm). Picric acid (Nº. 1) 0.5 mL 0.5 mL
Absorbance will be A1.
Mix and incubate in a water bath at 37 ºC during 10 minutes. Measure the

02 English - Ref.: 35
 2 2
absorbance of the Unknown and Standard against Blank at 510 nm or 60 mL/min/1.73m or > 60 mL/min/1.73m .
green filter (500 - 540 nm).
Calibration . Standard is traceable to the Standard Reference Material
Absorbance will be A1. (SRM) 914 of the National Institute of Standards and Technology (NIST).
Acid solution (Nº. 4) 0.1 mL 0.1 mL
Manual calibrations
Mix and let at room temperature for 5 minutes. Measure the absorbance of Perform a new calibration after reagent lot change or when the internal
the Unknown against Blank at 510 nm or green filter (500 - 540 nm). quality control indicates.

Absorbance will be A2. Quality control . For quality control use Qualitrol H Level 1 and
Use the same calculation proposed for the direct assay. Do not apply the
Qualitrol H Level 2 or other suitable control material. The limits and control
correction index.
interval must be adapted to the laboratory requirements.
In turbid samples in excess, it is not possible to get a clear supernatant in
Each laboratory should establish corrective actions to be taken if values
the deproteinization procedure. In this case, it is not possible to measure
fall outside the control limits.
the creatinine.

Endogenous creatinine depuration. The laboratory must Measurement/reportable range

inform the patient how to collect the correct urine within 24 hours. The reaction is linear between 0.2 mg/dL and 12 mg/dL.
Use the proposed methodologies to measure creatinine in serum and If creatinine concentration exceeds 12 mg/dL, the sample must be diluted
urine. with 0.85% NaCl. Multiply the result by the appropriate dilution factor.
Apply the obtained results in the following equation:
Expected values . Each laboratory should evaluate the
U transferability of the expected range to its own patient population and, if
8,10
Depuration (mL/minute) = x VM necessary, estimate its own reference range .
S
Serum/Plasma (mg/dL)*
U: Creatinine in urine (mg/dL)
newborn 0.31 - 0.92
S: Creatinine in serum (mg/dL)
VM: Volume per minute (urinary volume of 24 hours, in mL, divided by 2 weeks - 1 year 0.16 - 0.39
1440). 1 - <3 years 0.17 - 0.35
PS: Depuration must be corrected to the patient's body surface that is 3 - <5 years 0.26 - 0.42
obtained by a nomogram correlating weight and height, or using the 5 - < 7 years 0.29 - 0.48
following equation: 7 - <9 years 0.34 - 0.55
0.425 0.725
A = W x H x 0.007184 9- <11 years 0.32 - 0.64
11 - <13 years 0.42 - 0.71
2
A = body surface (m ) 13 - <15 years 0.46 - 0.81
W = weight (kg) Adults (women) 18 - 74 0.53 - 1.00
H = height (cm) Adults (men) 18 - 74 0.70 - 1.20

Multiply the depuration value by 1.73 and divide by the patient body * Corrected values with the correction index and IDMS traceable.
surface.
There isn't established range for the age 15 - <18 years. It's suggested to
Glomerular de filtration rate . The NKDEP4 recommends that the use the women and men adult range.
laboratories report the estimated glomerular filtration rate (eGFR) in all the
reports containing creatinine results. Conversion mg/dL to IS Units: mmol/L = mg/dL x 88.4.
When the plasmatic creatinine results are corrected and traceable
to the IDMS, the following equations that apply creatinine (CREA), age Urine (mg/kg/24 hours)
(18 - 70 years) and sex are used. 2 - 3 years 6 - 22
>3 years 12 - 30
Women Adults (women) 16 - 22
-0.203
eGFR (mL/min/1.73m2) = 175*(CREA)-1.154*(Age) *0.742 Adults (men) 21 - 26
2
Men Creatinine Depuration (mL/minute/1,73 m )**
eGFR (mL/min/1.73m2) = 175*(CREA)-1.154*(Age)-0.203 Children 70 - 140
Adults (women) 88 - 128
4
According NKDEP recommendations, the eGFR must be reported as the Adults (men) 97 - 137
2
calculated value when the result is £60 mL/min/1.73m . When the
calculated value is higher than 60, must be reported as: higher than ** Established values for not corrected and not IDMS traceable results

03 English - Ref.: 35
 4
The NKDEP recommends calculate the glomerular filtration rate (eGFR) 3. It is suggested to consult “www.fxol.org/” in order to review
in substitution of Creatinine Depuration, using the creatinine IDMS physiopathological source and drugs interference in results and
traceable result after application of the correction index. methodology.

Performance characteristics References

Recovery studies . In two samples with creatinine concentrations of 1. Cook JGH. Clin Chim Acta 1971;32:485-6.
1.2 and 3.2 mg/dL were added different quantities of the analyte.
Subsequent analyses provided recoveries ranging from 93 to 98%. The 2. Yatzidis H. Clin Chem 1974;20:1131-34.
mean proportional systematic error at 3.0 mg/dL decision level was
3. Spencer K. Ann Clin Biochem 1986;23:1-25.
0.1 mg/dL or 3.7 %.
4. Meyers GL, Miller WG, Coresh J et al. Clin Chem 2006;52:5-18.
Method comparison . A group of 20 sera were assayed by the
Creatinine method and the Creatinine K method (traceable to IDMS 5. Heinegard D, Tiderström G. Clin Chim Acta 1973;43:305-10.
method). Serum creatinine values ranged from 0.59 - 4.40 mg/dL. The
comparisons yielded a correlation coefficient of 0.995 and regression 6. Sociedad Española de Bioquímica Clínica y Patologia Molecular,
equation was 1.003x + 0.00. The mean total systematic error Base de Datos de Variación Biológica. Avaiable in :
(proportional and constant) at 1.00 mg/dL, 1.20 mg/dL and 2.00 mg/dL <htttp://www.seqc.es/ar ticle/ar ticleview/330/1/170> (access
decision levels were 0.003 (0.30 %), 0.004 (0.30 %) and 0.006 (0.30 %), 2006/04).
respectively.
7. Basques JC. Especificações da Qualidade Analítica. Labtest
Diagnóstica 2005.
Imprecision - Within Run
8. Junge W, Wilke B, Halabi A, Klein G. Clin Chim Acta 2004;344:137-48.
Mean SD
N (%) CV
(mg/dL) (mg/dL) 9. Martensson A, Rustad P, Lund H, Ossowicki H. Scand J Clin Lab Invest.
Sample 1 20 2.0 0.04 2.0 2004;64:439-42.
Sample 2 20 2.8 0.04 1.3
10. Ceriotti F, Boyd JC, Klein G et al. Clin Chem 2008; 54:559-66.

11. Labtest: Data on file.

Imprecision - Run-to-Run
Mean SD
N (%) CV
(mg/dL) (mg/dL) Presentation
Sample 1 20 2.0 0.05 2.5
Sample 2 20 2.8 0.06 2.3 Product Reference Contents
1 X 50 mL
1 X 200 mL
35-100
Analytical sensitivity . Detection limit: 0.3 mg/dL. The detection 1 X 10 mL
limit represents the lowest measurable creatinine concentration that can 1 X 10 mL
Creatinine
be distinguished from zero. It is calculated as two standard deviations of 1 X 250 mL
20 replicates of one sample without creatinine. 1 X 1000 mL
35E-500
1 X 30 mL
Matrix dilution effects . Two samples with values equal of 16.5 1 X 50 mL
and 18.6 mg/dL were used to evaluate the system response on the matrix
dilutions with 150 mmol/L NaCl (0.85%). Recoveries were found a range See availability of applications with Customer Service.
of 103 and 104 %, using dilution factors that vary from 2 to 4.

Notes

1. The material cleaning and drying are fundamental factors to the Customer information
reagent stability and to obtain correct results. [Warranty conditions]

2. The deionized or distilled water in the laboratory to prepare reagents, Labtest Diagnóstica warrants the performance of this product under the
use in the measurements and for final glass washing must have resistivity specifications until the expiration date shown in the label since the
³1 megaohm.cm, or conductivity £1 microsiems/cm and silicates application procedures and storage conditions, indicated on the label and
concentration must be <0.1mg/L. in this insert, have been followed correctly.

04 English - Ref.: 35
 Labtest Diagnóstica S.A.
CNPJ: 16.516.296 / 0001 - 38
Av. Paulo Ferreira da Costa, 600 - Vista Alegre - CEP 33400-000
Lagoa Santa . Minas Gerais Brasil - www.labtest.com.br
Customer Service email: customerservice@labtest.com.br

Edition: August, 1994 Copyright by Labtest Diagnóstica S.A.

Revision: September, 2020 Reproduction under previous autorization
Ref.: 290920

05 English - Ref.: 35
 Símbolos utilizados com produtos diagnósticos in vitro
Símbolos usados con productos diagnósticos in vitro
Symbols used with ivd devices

Conteúdo suficiente para < n > testes Risco biológico

Contenido suficiente para < n > tests Riesgo biológico
Contains sufficient for < n > tests Biological risk

Data limite de utilização (aaaa-mm-dd ou mm/aaaa) Marca CE

Estable hasta (aaaa-mm-dd o mm/aaaa) Marcado CE
Use by (yyyy-mm-dd or mm/yyyy) CE Mark

Material Calibrador Tóxico

Material Calibrador Tóxico
Calibrator Material Poison

Material Calibrador Reagente

Material Calibrador Reactivo
Calibrator Material Reagent

Limite de temperatura (conservar a) Fabricado por

Temperatura limite (conservar a) Elaborado por
Temperature limitation (store at) Manufactured by

Representante Autorizado na Comunidade Europeia Número do lote

Representante autorizado en la Comunidad Europea Denominación de lote
Authorized Representative in the European Community Batch code

Consultar instruções de uso Controle

Consultar instrucciones de uso Control
Consult instructions for use Control

Número do catálogo Controle negativo

Número de catálogo Control negativo
Catalog Number Negative control

Adições ou alterações significativas Controle positivo

Cambios o suplementos significativos Control positivo
Significant additions or changes Positive control

Produto diagnóstico in vitro Controle

Dispositivo de diagnóstico in vitro Control
In vitro diagnostic device Control

Liofilizado Corrosivo
Liofilizado Corrosivo
Lyophilized Corrosive

Período após abertura Uso veterinário

Período post-abertura Uso veterinario
Period after-opening Veterinary use

Instalar até
Instalar hasta
Install before Ref.: 140214

06 English - Ref.: 35